Research Article The Renal Protective Effect of Jiangya ...downloads.hindawi.com/journals/ecam/2015/428106.pdfHypertensive nephrosclerosis is one of the most common and serious chronic

Research ArticleThe Renal Protective Effect of Jiangya Tongluo Formulathrough Regulation of Adrenomedullin and Angiotensin II inRats with Hypertensive Nephrosclerosis

Lin Han1 Yan Ma2 Jian-guo Qin3 Li-na Li1 Yu-shan Gao1 Xiao-yu Zhang3 Yi Guo3

Lin-mei Song1 Yan-ni Luo3 and Xiao-yi Chi3

1College of Basic Medicine Beijing University of Chinese Medicine Beijing 100029 China2Molecular Research in Traditional Chinese Medicine Group Department of Pathophysiology and Allergy ResearchCenter of Pathophysiology Infectiology amp Immunology Vienna General Hospital Medical University of ViennaWaehringer Guertel 18-20 1090 Vienna Austria3Department of Nephrology Dongfang Hospital The Second Clinical Medical College of Beijing University of Chinese MedicineBeijing 100078 China

Correspondence should be addressed to Jian-guo Qin qindoctor163com

Received 13 December 2014 Revised 23 February 2015 Accepted 12 March 2015

Academic Editor Zhang Tan

Copyright copy 2015 Lin Han et alThis is an open access article distributed under the Creative Commons Attribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

We investigated the effect of Jiangya Tongluo (JYTL) formula on renal function in rats with hypertensive nephrosclerosis A totalof 21 spontaneously hypertensive rats (SHRs) were randomized into 3 groups valsartan (10mgkgd valsartan) JYTL (142 gkgdJYTL) and a model group (5mLkgd distilled water) Wistar Kyoto rats comprised the control group (119899 = 7 5mLkgd distilledwater) Treatments were administered by gavage every day for 8 weeks Blood pressure 24-h urine protein pathological changes inthe kidney serum creatinine and blood urea nitrogen (BUN) levels were estimated The contents of adrenomedullin (ADM) andangiotensin II (Ang II) in both the kidney and plasma were evaluated JYTL lowered BP 24-h urine protein serum creatinine andBUN ADM content in kidneys increased and negatively correlated with BP while Ang II decreased and negatively correlated withADM but there was no statistically significant difference of plasma ADM between the model and the treatment groups Possiblyactivated intrarenal renin-angiotensin system (RAS) plays an important role in hypertensive nephrosclerosis and the protectivefunction of ADM via local paracrine JYTL may upregulate endogenous ADM level in the kidneys and antagonize Ang II duringvascular injury by dilating renal blood vessels

1 Introduction

Hypertensive nephrosclerosis is one of the most commonand serious chronic complications in primary hypertensionIt is an independent risk factor for end-stage renal disease(ESRD) [1] approximately 25 of patients with hypertensivenephrosclerosis require a kidney transplant [2] Data from theUnited States Renal Data System suggest that hypertension isthe second cause of ESRD and the multiple risk factor inter-vention study revealed that approximately 49 of ESRD casesare caused by hypertension By 2002 there were 160 millionpatients with hypertension in China and approximately 96of patients were on dialysis as a result of renal artery sclerosis

due to hypertension Improvements in the early preventionand treatment of hypertension have resulted in significantreductions in mortality from cardiovascular complicationscaused by hypertension however for hypertensive nephro-sclerosis the mortality rates have not significantly improved[2] Therefore research efforts are now being focused onthe mechanism and prevention of hypertensive nephroscle-rosis Currently the treatment of hypertensive nephroscle-rosis involves controlling blood pressure and preglomeru-lar arteriolar resistance reducing intraglomerular pressureand improving renal ischemia However this approachhas failed to show a curative effect in approximately 50of patients and the application of angiotensin-converting

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2015 Article ID 428106 9 pageshttpdxdoiorg1011552015428106

2 Evidence-Based Complementary and Alternative Medicine

enzyme inhibitor (ACEI) drugs is limited when renal func-tion is impaired beyond a specific threshold (serum creati-nine gt3mgdL) Therefore further research on new drugsand treatment approaches to delay the progression of hyper-tensive nephrosclerosis towards end-stage renal failure isrequired

Adrenomedullin (ADM) is a vasodilator peptide that wasoriginally isolated from the extract of human pheochromocy-toma in 1993 It has numerous biological effects and is presentin a variety of tissues and organs particularly in the heartkidney and lungs [3] ADM is mainly produced by vascularendothelial and smoothmuscle cells whilemesangial and epi-thelial cells also secrete ADM [4] When combined with cal-citonin receptor-like receptors (CRLR) and receptor activity-modifying proteins 2 and 3 (RAMP2 and RAMP3) ADMhasa strong vasodilator effect [5] it increases renal blood flowand glomerular filtration via the expansion of efferent andafferent arterioles [4] ADM exerts important vascular andrenal protective effects through its interaction with Ang IIand other vasoconstrictive substances [6]Therefore ADM isconsidered important in preventing high blood pressure andhypertension-induced organ damage Consequently inves-tigation of the potential renal protective effects of ADMhas become a research focus in the field of hypertensionnephrosclerosis

We routinely use Jiangya Tongluo (JYTL) at our dailyclinic to treat hypertensive nephrosclerosis and we haveobserved good clinical curative effect To gain insight into themechanisms of regulation of JYTL in hypertensive nephroscle-rosis we investigated the effects of JYTL on blood pressurerenal function and the expression of ADM and Ang II inkidney and plasma using a spontaneous hypertensive rat(SHR) experimental model to explore its decompression andprotective effects on kidney

2 Materials and Methods

21 Animal Models and Drugs Sixteen-week-old male SHRs(mean weight = 200 plusmn 10 g) were purchased from VitalRiver Laboratories (Beijing China number SCXK 2002-2003) The animals were housed in the Central Laboratory atBeijingUniversity of ChineseMedicine with a 12-h lightdarkcycle and free access to food and water The SHRs werefed a normal diet for a week and were then randomized toone of 3 groups model (5mLkgd distilled water by gav-age) valsartan (10mgkgd valsartan by gavage) and JYTL(142 gkgd JYTL by gavage) The control group comprisedWistar Kyoto rats (5mLkgd distilledwater by gavage)Thesedoses were determined from a previous pharmacodynamicexperimental study [7] Each group was then sacrificed afteran 8-week treatment period All experimental procedureswere conducted in accordance with the guidelines for theuse of experimental animals and approval was granted bythe Institutional Review Committee on Animal Care and Useat the Experimental Animal Centre at Beijing University ofChineseMedicine (Certificate of Conformity SCXK Beijing2012-0001)

211 Preparation of JYTL Decoction JYTL is frequently usedin clinical practice for treatment of hypertension and iscomposed of 30 g Nacre (Zhen Zhu Mu ZZM) 15 g Cassiaoccidentalis (Cao Jue Ming CJM) 12 g Safflower (HongHua HH) 20 g Salvia miltiorrhiza (danshen DS) and 15 gChrysanthemum (juhua JH) The medicinal herbs wereprovided by Pharmacy Department at Dongfang HospitalBeijing China The herbs were first cut and boiled togetherin 6x volume of water for 05 h (first extraction) Residuefrom the first extraction was boiled in 8x volume of waterfor 25min Finally the filtered solutions were combined andconcentrated into an aqueous extract containing 12 gmL rawherbs Valsartan was purchased from Novartis Pharma Ltd(Beijing China)

22 Blood Pressure Detection Systolic blood pressure (BP)was monitored prior to treatment (0 weeks) and after 2 4 6and 8 weeks of treatment using a noninvasive computerizedtail-cuff system (BESN-II Desai Production BiotechnologyCo Nanjing China) and performed by specifically assignedinvestigators at regular intervals in order to minimize errorThe mean value from 3 measurements of BP was taken foreach rate representing the sample systolic pressure

23 Pathological Examination of Renal Tissue After 8 weeksof treatment rats were sacrificed under chloral hydrate anes-thesia (35 gkg administered via intraperitoneal injection)Blood was rapidly sampled by abdominal aorta puncture andthe serum was stored at minus80∘C prior to analysis Renal tissuewas excised washed in physiological saline snap-frozen inliquid nitrogen and then stored atminus80∘CKidney tissueswerefixed with 4 paraformaldehyde embedded in paraffin andcut into 3-120583m thick sections Hematoxylin-eosin (HE) stain-ing was performed in order to assess glomerular and vascularinjuries and perivascular lesions such as fibrosis under anelectronic scanner (SIPNOMIC 01579 Zeiss Co Germany)

24 Determination of Urinary Protein over 24 Hours Priorto and after 8 weeks of treatment the rats were placed inmetabolic cages where food was not provided (but water wasavailable) Urine samples were collected over 24 h to calculatethe total urinary output and 5mL was collected for centrifu-gation the supernatant was stored at minus20∘C A commercialradioimmunoassay (RIA) kit (PLA Institute of RIA BeijingChina)was used to estimate urine protein over 24 h in accord-ance with the manufacturerrsquos protocol

25 Determination of Serum Creatinine and Blood UreaNitrogen Levels After 8 weeks of treatment blood wascollected from the abdominal aorta and serum was separatedby centrifugation (1500timesg) at 4∘C for 10min and stored inEppendorf tubes Blood urea nitrogen (BUN) and creatininelevels were determined by the Jaffe and diacetyl-oximemethods in accordance with the manufacturer protocol

26 Determination of Ang II and ADM Content in the Kidneyand Plasma After 8 weeks of treatment rats were sacrificedand blood was sampled from the abdominal aorta Freshkidney tissues were collected weighed quickly ground with

Evidence-Based Complementary and Alternative Medicine 3

normal saline and boiled in water at 100∘C for 10min Thehomogenate was centrifuged (1500timesg) at 4∘C for 15min andthe supernatant was preserved at minus20∘C Ang II and ADMcontent in the renal tissue and plasma was determined usingan RIA Kit (PLA Institute of RIA Beijing China) in accor-dance with the manufacturerrsquos protocol The protein contentof the supernatant was also simultaneously determined forcorrection

27 Statistical Analysis All data were analyzed using SPSS160 software (SPSS Inc Chicago USA) Statistical signif-icance was analyzed using a one-way analysis of variancefollowed by the post hoc Student-Newman-Keuls multiplecomparison test All data are expressed as mean plusmn standarderror of the mean (SEM) and 119875 lt 005 was consideredstatistically significant

3 Results

31 Effect of JYTL on Blood Pressure Animal models in thisstudy comprised SHRswith knocked out genes Prior to treat-ment the BP of rats in the model group and both treatmentgroups was significantly higher than in the control group(119875 lt 001) (Figure 1(a)) the model and treatment groupswere comparable indicating that the model was successfullyestablished No decrease in BP of rats was observed in thevalsartan and JYTL groups compared with the model groupafter 2 weeks of treatment (Figure 1(b)) However at 4 6 and8 weeks of treatment the BP of rats in the JYTL and valsartangroups was significantly lower than that in the model group(119875 lt 001 and 119875 lt 005) Furthermore at 4 weeks theBP of rats in the valsartan group was lower than that in theJYTL group (Figure 1(c)) while at 6 and 8 weeks there wasno statistically significant differences between the two groups(Figures 1(d) and 1(e))

32 Effects of JYTL on Renal Pathological Morphology Weobserved severely pathological lesion in SHRs by hematoxy-lin-eosin staining It comprised glomerular ischemia andsclerosis tubular atrophy and hyaline degeneration andinterstitial fibrosis with inflammatory cells hyperplasiaand the interlobular arteries show intimal thickening(Figure 2(b)) Such lesions were considerably diminishedby JYTL and valsartan the pathological change representedinterlobular artery mild-thickening with tubular vacuolardegeneration (Figure 2(c)) and tubular ectasia with hyalinedegeneration (Figure 2(d)) Furthermore the renal structureof Wistar Kyoto rats appeared normal (Figure 2(a)) Thisindicated that JYTL and valsartan could significantly protectthe kidney from hypertension-induced lesions This resultstrongly supported the therapeutic potential of JYTL againsthypertensive nephrosclerosis

33 Effects of JYTL onUrinary ProteinQuantity over 24HoursThe quantity of urinary protein over 24 h was statisticallysignificantly higher in the model group and both treatmentgroups compared with the control group (119875 lt 001) therewas no statistically significant difference between modeland treatment groups indicating a successful model After

8 weeks of treatment the 24-h urine protein level wassignificantly reduced in both themodel and treatment groups(119875 lt 001) it was slightly lower in the JYTL group thanin the valsartan group however these differences were notstatistically significant (Figures 3(a) and 3(b))

34 Effects of JYTL on Renal Function Following 8 weeks oftreatment serum creatinine levels were significantly reducedin both JYTL and valsartan groups (119875 lt 001) serumcreatinine levels in JYTL groupwere slightly lower than in thevalsartan group however there was no statistically significantdifference between the two groups (119875 gt 005) The bloodurea nitrogen content was significantly reduced in both JYTLand valsartan groups after the 8-week treatment period (119875 lt001) While BUN was lower in the JYTL group than in thevalsartan group this difference was statistically significant(119875 lt 005) (Figures 4(a) and 4(b))

35 Effects of JYTL on the Expression of Ang II and ADM in theKidneys and ADM in Plasma The results of RIA showed thatkidney Ang II expression in the model JYTL and valsartangroups was statistically significantly higher than that in thecontrol group (119875 lt 001) Ang II expression was statisticallysignificantly lower in the valsartan and JYTL groups thanin the model group (119875 lt 001) While Ang II expressionin valsartan group was slightly higher than the JYTL groupthese differences were not statistically significantly different(Figure 5(a)) Levels of ADM in the model JYTL andvalsartan kidneys were statistically significantly lower thanthe control (119875 lt 001) and significantly higher in thevalsartan group compared with the model (119875 lt 001)While the level of ADM in the valsartan group was slightlyhigher than that in the JYTL group this difference wasnot statistically significant (Figure 5(b)) After 8 weeks oftreatment the content of Ang II in plasma in themodel groupwas statistically significantly higher than that in the controlgroup (119875 lt 001) In treatment groups though the levels ofAng II were higher than that of control group but there wasno statistically significant difference between them and alsothere was no statistically significant difference between themodel and treatment groups (Figure 5(c)) Meanwhile thecontent of ADM in the plasma in the model group and bothtreatment groupswas statistically significantly lower than thatin the control group (119875 lt 001 and 119875 lt 005 resp) but therewas no statistically significant difference between the modeland treatment groups (Figure 5(d))

36 Correlation of ADM with Ang II and BP There was astatistically significant negative correlation between ADMandAng II and between ADMand BP in the kidneys (Figures6(a) and 6(b))

4 Discussion

Hypertensive nephrosclerosis is a form of hypertension-induced arteriolar nephrosclerosis and it results from benignarteriolar nephrosclerosis Pathological changes indicative of


0

50

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200

250

Control Model Valsartan JYTL

Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast

lowast

(a)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowast lowastlowast

(b)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowast lowast998779amplowast

(c)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast lowast998779

(d)

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100

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200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast998771lowast

(e)

Figure 1 Effect of JYTL on mean plusmn SEM blood pressure (119899 = 7 per study group) (a) Before treatment (b) After 2 weeks of treatment (c)After 4 weeks of treatment (d) After 6 weeks of treatment (e) After 8 weeks of treatment lowast119875 lt 001 model valsartan and JYTL rats versuscontrol rats 119875 lt 001 rats in the valsartan group versus model rats 119875 lt 005 and 998771119875 lt 001 rats in the JYTL group versus model ratsamp119875 lt 005 rats in the JYTL group versus valsartan group

the condition include renal afferent arteriole hyaline degen-eration and myointimal hypertrophy in the arteria inter-lobulares and arteria arcuata [8 9] These lead to ischemicchanges in the glomeruli and interstitium and consequentlycompromise renal function [10]

Decompression is the modern universally accepted formof hypertensive nephrosclerosis therapy Studies have demon-strated a significant contribution of ACE I and Ang II type1 receptor blockers (ARB) in the prevention of renal andcardiovascular damage [11 12] However this form of treat-ment has failed to demonstrate an obvious curative effect inapproximately 50 of patients Furthermore the vasodilator

effect of ACEI and ARB on efferent arterioles is superiorto afferent arterioles that impacts the compensatory mech-anisms of intraglomerular pressure hypertransfusion andhyperfiltration and accelerates renal function aggravationFor those patients with renal insufficiency administeringACEI frequently predisposes them to hyperkalemia There-fore when renal functional impairment crosses a specificthreshold (serum creatinine gt3mgdL) the application ofACE I and ARB is limited Consequently it has becomeimportant to explore new therapeutic methods to delay theprogression of hypertensive nephrosclerosis towards end-stage renal disease


200120583m

(a)

200120583m

(b)

200120583m

(c)

200120583m

(d)

Figure 2 Histologic studies of JYTL in hypertensive nephrosclerosis rats after HE staining (a) Control group (b) Model group (c) Valsartangroup (d) JYTL group Bar = 200120583m samemagnification for all panels (a)The arrows point to the glomerular and the lined epithelia of renaltubules in normal condition (b) Glomerular ischemia and sclerosis (blue arrow) interlobular artery with marked myointimal proliferation(yellow arrows) tubular atrophy and hyaline degeneration (little black arrow) and interstitial fibrosis with inflammatory cells hyperplasia(black arrow) (c) Interlobular artery mild-thickening with tubular vacuolar degeneration (d) Tubular ectasia with hyaline degeneration HEin 3-120583m thick sections 200

0

100

200

300

400

500

600

700

800


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowastlowast

lowast

(a)

0100200300400500600700800900

1000


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowast

lowast998779lowast

(b)

Figure 3 Effect of JYTL on the mean plusmn SEM urine protein content over 24 h (119899 = 7 per study group) (a) Before treatment (b) After 8 weeksof treatment with JYTL lowast119875 lt 001 model valsartan and JYTL rats versus control rats 119875 lt 001 rats in the valsartan group versus modelrats 119875 lt 001 rats in the JYTL group versus model rats

Thepathogenesis and etiology of hypertensive nephroscle-rosis are complex The mechanism behind the condition is asfollows (1) Glomeruli hypertension results in vascular endo-thelial cell damage and increases vasoconstrictor (Ang IIET-1) and platelet derived growth factors and the synthesisand secretion of extracellular matrix [11 12] (2) Elevations inthe glomerular capillary pressure lead to an increase in some

of the transforming growth factors such as TGF-1205731 thisprocession stimulates collagen deposition and the prolifera-tion of mesangial cells and increases the extracellular matrixeventually leading to kidney sclerosis [13] (3)The glomerularischemia inflammatory response causes increases in vascularinjury vasoactive substances chemical chemokines andmitogenic factors and can aggravate kidney damage [14]


0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

Bloo

d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast

lowast998779amplowast

(b)

Figure 4 Effect of JYTL on mean plusmn SEM renal function (119899 = 7 in each group) (a) Serum creatinine (b) Blood urea nitrogen lowast119875 lt 001model valsartan and JYTL rats versus control rats 119875 lt 001 rats in the valsartan group versus model rats 119875 lt 001 rats in the JYTLgroup versus model rats amp119875 lt 005 JYTL versus valsartan group

0123456789


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Ang

II in

kid

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(pg

g)

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(a)

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05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

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250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540


lowastlowastlowast lowastlowast

Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)

Figure 5 Effect of JYTL on mean plusmn SEM ADM and Ang II content (119899 = 7 per group) (a) Ang II in kidney (b) ADM in kidney (c) Ang II inplasma (d) ADM in plasma lowast119875 lt 001 and lowastlowast119875 lt 005 model valsartan and JYTL rats versus control rats 119875 lt 001 rats in the valsartangroup versus model rats 119875 lt 001 JYTL group versus model group

Reactive oxygen species can stimulate enzymes which haveoxidative stress sensitivity and nuclear transcription factors(NF-120581B and AP-1) and can increase the expression levels ofcytokine and chemotactic and adhesion factors The finalstages of the mechanism of hypertensive nephrosclerosisinclude triggering renal interstitial inflammation renalfibroblasts proliferation and conversion and the formationof renal fibrosis [15]

The functional impairment of vascular endothelial cellstherefore plays a critical role in the pathogenesis of hyperten-sive nephrosclerosis Apoptosis of microvascular endothelialcells inducesmicrovessel rarefaction and increases peripheralresistance which in turn increases BP decreases the ability ofsubstance and energymetabolism and reduces reserve capac-ity and material exchange ability resulting in the injury oftarget organsThis present study found that Ang II promoted


r = minus0853

P lt 0001

n = 7

0123456789

10

0 1 2 3 4Content of ADM in kidney (pgg)

Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

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250

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Bloo

d pr

essu

re (m

mH

g)

(b)

Figure 6 (a) Correlation between ADM and Ang II levels in the kidney (b) Correlation between BP and ADM levels in the kidney

endothelial cell apoptosis in a concentration-dependentman-ner Endothelial cell apoptosis induces an imbalance ofproliferation versus apoptosis of smooth muscle cells Thisprocession eventually results in changes to the structure andfunction of blood vessels and target organ damage [16ndash18]Therefore inhibition of blood vessel injuries by Ang II is keyto the treatment of hypertension-induced organ injury

AMD is a novel vasorelaxant peptide It is critical for bothvascular remodeling and blood pressure regulation and playsan important role in the protection of kidneys and blood ves-sels [5] It dilates the blood vessels in two ways endothelium-dependent dilation and endothelium-independent dilation[19ndash21] ADM can increase blood flow and glomerularfiltration rate through the dilation of the afferent andefferent arterioles [4] It can also block the action of AngII-induced mitogen-activated protein kinase resulting inthe prohibition of overproliferation of smooth muscle cells[4 22] and the inhibition of endothelial cell apoptosisthrough a NO-dependent mechanism [23] ADM promotesimpaired endothelial cells reendothelialization through thecyclic adenosine monophosphate and phosphatidylinositol3-kinase pathways consequently playing a role in vascularendothelial growth and the promotion of angiogenesis [2425]

Collagen and elastin which are secreted by fibroblastsand inhibit vascular sclerosis through the regulation ofmatrix metalloproteinase-2 activity and protein expressionare reduced by ADM [26] Therefore ADM is regarded asa crucial factor not only for preventing an increase in BPbut also for preventing target organ damage Consequentlystudies of ADM have become a research focus in thefield of hypertension nephrosclerosis ADM is an endogenicsubstance involved in a variety of bodily functions in anautocrine and paracrine manner however it is expensiveand therefore unsuitable for long-term therapy Thereforethe development of drugs that promote the generation ofendogenous ADM and improve its bioactivity has become anurgent requirement

Traditional Chinesemedicine (TCM) has been used sincea long time and positive treatment effects have been observedin hypertensive nephrosclerosis [27 28] JYTL formula usedin this study is composed of Nacre Cassia occidentalisSafflower Salvia miltiorrhiza and Chrysanthemum Thispreparation was derived from the JYTM formula invented byShikui Guo (one of the most prestigious TCM cardiovascularspecialists) and it has obvious therapeutic effects on car-diovascular disease [29] Modern pharmacological researchhas shown that Nacre exerts an anti-inflammatory effectin the treatment of vascular headache Cassia occidentalisdepresses blood pressure reduces the extracellular matrixinhibits mesangial cell proliferation reduces proteinuria andimproves renal function Safflower and Salvia miltiorrhizahave antioxidant and anti-inflammatory effects clinical val-idation confirms their curative effect in terms of promotingblood circulation to remove blood stasisThey are also knownto play a major role in renal interstitial fibrosis vascularendothelial injury and oxidative stress [29ndash33] Chrysanthe-mum can cause significant dilation of the coronary arteryincrease in coronary bloodflow and improvement ofmyocar-dial cells for hypoxia tolerance It also plays an importantrole in decompression and often synergizes with other drugsto reduce hypertension [34] In this study we used JYTL totreat hypertensive nephrosclerosis in a rat model and explorethemechanisms underlying its decompression and protectiveeffects on kidney function

The results demonstrated that after 4 weeks of treatmentwith JYTL the BP of rats decreased though valsartan groupshowed better results than the JYTL group at this timeHowever with prolonged JYTL treatment at 6 and 8 weeksJYTL group showed similar antihypertensive effect as val-sartan Since there are no obvious medical contraindicationsto TCM JYTL also can be used when renal functionalimpairment crosses a specific threshold (serum creatininegt3mgdL) On the other hand valsartan causes reducedglomerular filtration rate (GFR) aggravates renal injuryand is not recommended to use under such conditions


Our results also showed that JYTL reduced albuminuriaimproved renal function and alleviated renal pathologicaldamage in SHR To further explore the mechanism of actionof JYTL we estimated the content of Ang II and ADM inthe kidneys and plasma We observed that the plasma ADMcontent decreased rapidly in the 24-week-old SHR which isdifferent from previous research [35 36] This may be dueto progressive endothelial cell injury and the self-regulationfunction in a state of decompensation However valsartanand JYTL showed no effect on the level of ADM and Ang IIin plasma which is generally comparable to previous studies[37 38] Some authors have described high levels of Ang IIin kidney tissue [39] and locally generated Ang II has beenimplicated in paracrine regulatory mechanisms leading toaltered proliferative and synthetic responses of cells [40]Therefore we speculated that the renal protective effect ofADM is not associated with circulating ADM and perhapsrenal local RAS plays a more important role in the processof hypertensive nephrosclerosis We explored the correlationbetween ADM and Ang II in the kidneys The data showedthat ADM content in SHR kidneys decreased significantlyas compared to that in control rats while Ang II increasedADM protein content showed a statistically significant neg-ative correlation between BP and Ang II levels JYTL andvalsartan upregulated the level of ADM and downregulatedAng II level in the kidneyThus it can be concluded thatADMmay act as a paracrine factor affecting renal function duringhypertensive nephrosclerosis and the renal protective effectof JYTL may occur via upregulation of endogenous ADMlevels in the kidney and antagonism of vascular injury by AngII Further research is required to address the effect of JYTLon receptor systems and the signal transduction pathway ofADM In addition further investigations on the molecularmechanism of hypertensive nephrosclerosis may providealternative therapy and new drug screening approaches

5 Conclusion

JYTL has renal protective effect in SHRs that may be relatedto the upregulation of ADM levels in the kidney and theinhibition of vascular injury

Conflict of Interests

The authors declare that they have no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Lin Han and Yan Ma equally contributed to this paper Jian-guo Qin obtained funding and supervised this study LinHan conducted the experiments conceived and designedthe experiments and wrote the draft of the paper Yan Madesigned this study and participated in writing of the paperLi-na Li and Yu-shan Gao provided technical support Xiao-yu Zhang Yi Guo Lin-mei Song Yan-ni Luo and Xiao-yiChi helped perform the animal experiments and LinHan andJian-guo Qin analyzed the data

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (Grant no 81173407) and the Admin-istration of Traditional Chinese Medicine of Beijing for theShikui Guo Famous Doctor Research Lab Project to JianguoQin (no 2011-SZ-A-27)

References

[1] A J Collins R N Foley B Chavers et al ldquoUnited StatesRenal Data System 2011 Annual Data Report atlas of chronickidney disease amp end-stage renal disease in the United StatesrdquoAmerican Journal of Kidney Diseases vol 59 no 1 supplement1 p A7 2012

[2] B Tomas ldquoAmerican Society of Nephrology renal researchreportrdquo Journal of the American Society of Nephrology vol 16no 7 pp 1886ndash1903 2005

[3] A Cases and JMora-Macıa ldquoAdrenomedullin a new vasoactivepeptiderdquo Nefrologia vol 21 no 1 pp 16ndash25 2001

[4] J Kato T Tsuruda T Kita K Kitamura and T Eto ldquoAdren-omedullin a protective factor for blood vesselsrdquoArteriosclerosisThrombosis and Vascular Biology vol 25 no 12 pp 2480ndash24872005

[5] J Bełtowski and A Jamroz ldquoAdrenomedullinmdashwhat do weknow 10 years since its discoveryrdquo Pharmacology vol 56 no1 pp 5ndash27 2004

[6] C-Y Ma T-G Cui and C-S Tang ldquoProtective effect of adren-omedullin on human renal tubulointerstitial lesionrdquo BasicMedical Sciences and Clinics vol 21 no 2 pp 151ndash153 2001

[7] J-G Qin L Han Y-H Wang et al ldquoAction mechanismof Tongmai Yishen Fang in antagonism of renal interstitialfibrosisrdquo Journal of Beijing University of Traditional ChineseMedicine vol 35 no 11 pp 739ndash742 2012

[8] S-Q Li X-M Liu and W-Z Wang ldquoThe research studyof Salvia miltiorrhiza effects on hypertension nephrosclerosisratsrdquo Chinese Archives of Traditional Chinese Medicine vol 24no 2 pp 224ndash225 2006

[9] W-Z Zou ldquoInstruction on pathologic diagnostic criteria ofrenal biopsyrdquo Chinese Journal of Nephrology vol 17 no 4 pp270ndash275 2001

[10] I Mimura andM Nangaku ldquoThe suffocating kidney tubuloin-terstitial hypoxia in end-stage renal diseaserdquo Nature ReviewsNephrology vol 6 no 11 pp 667ndash678 2010

[11] ldquoEffect of enalapril on mortality and the development of heartfailure in asymptomatic patients with reduced left ventricularejection fractionsrdquo The New England Journal of Medicine vol327 no 10 pp 685ndash691 1992

[12] B M Brenner M E Cooper D de Zeeuw et al ldquoEffects oflosartan on renal and cardiovascular outcomes in patients withtype 2 diabetes and nephropathyrdquo The New England Journal ofMedicine vol 345 no 12 pp 861ndash869 2001

[13] D Fliser ldquoPerspectives in renal disease progression the endo-thelium as a treatment target in chronic kidney diseaserdquo Journalof Nephrology vol 23 no 4 pp 369ndash376 2010

[14] H-T Kuo S-J Shin M-C Kuo and H-C Chen ldquoEffects ofspecific endothelin-1 receptor antagonists on proliferation andfibronectin production of glomerularmesangial cells stimulatedwith angiotensin IIrdquo Kaohsiung Journal of Medical Sciences vol22 no 8 pp 371ndash376 2006


[15] H T Tang D S Cheng Y T Jia et al ldquoAngiotensin II inducestype I collagen gene expression in human dermal fibrovlaststhrough anAP-1 TCF-1205731-dependent pathwayrdquoBiochemical andBiophysical Research Communications vol 385 no 3 pp 418ndash423 2009

[16] P Minuz P Patrignani S Gaino et al ldquoIncreased oxidativestress and platelet activation in patients with hypertension andrenovascular diseaserdquo Circulation vol 106 no 22 pp 2800ndash2805 2002

[17] A J Polichnowski C Jin C Yang and A W Cowley ldquoRoleof renal perfusion pressure versus angiotensin II on renaloxidative stress in angiotensin II-induced hypertensive ratsrdquoHypertension vol 55 no 6 pp 1425ndash1430 2010

[18] N Bouvier J P Flinois J Gilleron et al ldquoCyclosporine triggersendoplasmic reticulum stress in endothelial cells a role forendothelial phenotypic changes and deathrdquo The AmericanJournal of PhysiologymdashRenal Physiology vol 296 no 1 ppF160ndashF169 2009

[19] P-X Xu H Li and Z-Y Sun ldquoExpression of vascular endothe-lial growth factor in myocardium of spontaneously hyperten-sive ratsrdquo Medical Journal of the Chinese Peoplersquos vol 16 no 7pp 491ndash493 2005

[20] S-L Wang and Z-B Zhang ldquoApoptosis of vascular endothelialcellsrdquo Journal of Liaoning University of TCM vol 13 no 3 pp50ndash53 2011

[21] F Yoshihara S-I Suga N Yasui et al ldquoChronic administrationof adrenomedullin attenuates the hypertension and increasesrenal nitric oxide synthase in Dahl salt-sensitive ratsrdquo Regula-tory Peptides vol 128 no 1 pp 7ndash13 2005

[22] Y Shimekake K Nagata S Ohta et al ldquoAdrenomedullin stim-ulates two signal transduction pathways cAMP accumulationand Ca2+ mobilization in bovine aortic endothelial cellsrdquo TheJournal of Biological Chemistry vol 270 no 9 pp 4412ndash44171995

[23] T Horio M Kohno H Kano et al ldquoAdrenomedullin as anovel antimigration factor of vascular smooth muscle cellsrdquoCirculation Research vol 77 no 4 pp 660ndash664 1995

[24] M Kohno K Yokokawa H Kano et al ldquoAdrenomedullin is apotent inhibitor of angiotensin II-induced migration of humancoronary artery smooth muscle cellsrdquoHypertension vol 29 no6 pp 1309ndash1313 1997

[25] H K Wong T T Cheung and B M Y Cheung ldquoAdrenom-edullin and cardiovascular diseasesrdquo JRSM Cardiovascular Dis-ease vol 1 no 5 article 14 7 pages 2012

[26] K Miyashita H Itoh N Sawada et al ldquoAdrenomedullinprovokes endothelial Akt activation and promotes vascularregeneration both in vitro and in vivordquo FEBS Letters vol 544no 1ndash3 pp 86ndash92 2003

[27] S EHermansen T Lund TKalstad K Ytrehus andTMyrmelldquoAdrenomedullin augments the angiogenic potential of lateoutgrowth endothelial progenitor cellsrdquo American Journal ofCell Physiology vol 300 no 4 pp C783ndashC791 2011

[28] T Tsuruda J Kato Y-N Cao et al ldquoAdrenomedullin inducesmatrix metalloproteinase-2 activity in rat aortic adventitialfibroblastsrdquo Biochemical and Biophysical Research Communica-tions vol 325 no 1 pp 80ndash84 2004

[29] X-X Zhu D Yan S-H Yan et al ldquoEmpirical study ofJiang ya Yi shen particles on ET-1 and TGF-120573

1

expression inspontaneously hypertensive rats (SHR)rdquo Chinese Archives ofTraditional ChineseMedicine vol 28 no 9 pp 1805ndash1806 2010

[30] I J Cho C Lee and T Y Ha ldquoHypolipidemic effect of solublefiber isolated from seeds of cassia tora Linn In rats fed a high-cholesterol dietrdquo Journal of Agricultural and Food Chemistryvol 55 no 4 pp 1592ndash1596 2007

[31] J Ma Z Chen and Y Wang ldquoThe clinical observation oftreatment of hypertensive renal injury edema by combinationof fangjihuangqi decoction and western medicinerdquo ChineseJournal of Basic Medicine in Traditional Chinese Medicine vol18 no 8 pp 879ndash880 2012

[32] H-Q Tao J Li J Zhang et al ldquoThe curative effect observationof treatment of early stage in hypertensive renal injury bycombination of Liuwei Dihuang Wan and western medicinerdquoShaanxi Journal of Traditional Chinese Medicine vol 33 no 4pp 411ndash412 2012

[33] W Zheng and S-Q Li ldquoThe effect of tongxinluo on blood vesselendothelium in spontaneously hypertension ratrdquo Chinese Jour-nal of Integrative Medicine on Cardio-Cerebrovascular Diseasevol 7 no 2 pp 179ndash180 2009

[34] Z Jan Y-B Li D-W Qian et al ldquoChrysanthemum chemicalcomponents and pharmacological activity research progressrdquoLishizhen Medicine and Materia Medica Research vol 17 no 10pp 1941ndash1942 2006

[35] R Tang Q-L Zhou P Veeraragoo Z Xiao and Z Liu ldquoEffectsof cordyceps sinensis on renal Klotho expression and oxidativestress in spontaneously hypertensive ratsrdquo Chinese Journal ofNephrology Dialysis amp Transplantation vol 19 no 4 pp 338ndash343 2010

[36] J-G Qin Y-HWang J-P Liang et al ldquoWeiqinGuorsquos treatmentexperience about hypertensionrdquo Journal of Traditional ChineseMedicine vol 48 no 7 pp 586ndash589 2007

[37] T Ishimitsu T Nishikimi Y Saito et al ldquoPlasma levels ofadrenomedullin a newly identified hypotensive peptide inpatients with hypertension and renal failurerdquo Journal of ClinicalInvestigation vol 94 no 5 pp 2158ndash2161 1994

[38] M Kohno T Hanehira H Kano et al ldquoPlasma adrenomedullinconcentrations in essential hypertensionrdquoHypertension vol 27no 1 pp 102ndash107 1996

[39] B Braam K D Mitchell J Fox and L G Navar ldquoProximaltubular secretion of angiotensin II in ratsrdquoTheAmerican Journalof PhysiologymdashRenal Fluid and Electrolyte Physiology vol 264no 5 part 2 pp F891ndashF898 1993

[40] M Bader J Peters O Baltatu D N Muller F C Luft and DGanten ldquoTissue renin-angiotensin systems new insights fromexperimental animal models in hypertension researchrdquo Journalof Molecular Medicine vol 79 no 2 pp 76ndash102 2001

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



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OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


enzyme inhibitor (ACEI) drugs is limited when renal func-tion is impaired beyond a specific threshold (serum creati-nine gt3mgdL) Therefore further research on new drugsand treatment approaches to delay the progression of hyper-tensive nephrosclerosis towards end-stage renal failure isrequired

Adrenomedullin (ADM) is a vasodilator peptide that wasoriginally isolated from the extract of human pheochromocy-toma in 1993 It has numerous biological effects and is presentin a variety of tissues and organs particularly in the heartkidney and lungs [3] ADM is mainly produced by vascularendothelial and smoothmuscle cells whilemesangial and epi-thelial cells also secrete ADM [4] When combined with cal-citonin receptor-like receptors (CRLR) and receptor activity-modifying proteins 2 and 3 (RAMP2 and RAMP3) ADMhasa strong vasodilator effect [5] it increases renal blood flowand glomerular filtration via the expansion of efferent andafferent arterioles [4] ADM exerts important vascular andrenal protective effects through its interaction with Ang IIand other vasoconstrictive substances [6]Therefore ADM isconsidered important in preventing high blood pressure andhypertension-induced organ damage Consequently inves-tigation of the potential renal protective effects of ADMhas become a research focus in the field of hypertensionnephrosclerosis

We routinely use Jiangya Tongluo (JYTL) at our dailyclinic to treat hypertensive nephrosclerosis and we haveobserved good clinical curative effect To gain insight into themechanisms of regulation of JYTL in hypertensive nephroscle-rosis we investigated the effects of JYTL on blood pressurerenal function and the expression of ADM and Ang II inkidney and plasma using a spontaneous hypertensive rat(SHR) experimental model to explore its decompression andprotective effects on kidney

2 Materials and Methods

21 Animal Models and Drugs Sixteen-week-old male SHRs(mean weight = 200 plusmn 10 g) were purchased from VitalRiver Laboratories (Beijing China number SCXK 2002-2003) The animals were housed in the Central Laboratory atBeijingUniversity of ChineseMedicine with a 12-h lightdarkcycle and free access to food and water The SHRs werefed a normal diet for a week and were then randomized toone of 3 groups model (5mLkgd distilled water by gav-age) valsartan (10mgkgd valsartan by gavage) and JYTL(142 gkgd JYTL by gavage) The control group comprisedWistar Kyoto rats (5mLkgd distilledwater by gavage)Thesedoses were determined from a previous pharmacodynamicexperimental study [7] Each group was then sacrificed afteran 8-week treatment period All experimental procedureswere conducted in accordance with the guidelines for theuse of experimental animals and approval was granted bythe Institutional Review Committee on Animal Care and Useat the Experimental Animal Centre at Beijing University ofChineseMedicine (Certificate of Conformity SCXK Beijing2012-0001)

211 Preparation of JYTL Decoction JYTL is frequently usedin clinical practice for treatment of hypertension and iscomposed of 30 g Nacre (Zhen Zhu Mu ZZM) 15 g Cassiaoccidentalis (Cao Jue Ming CJM) 12 g Safflower (HongHua HH) 20 g Salvia miltiorrhiza (danshen DS) and 15 gChrysanthemum (juhua JH) The medicinal herbs wereprovided by Pharmacy Department at Dongfang HospitalBeijing China The herbs were first cut and boiled togetherin 6x volume of water for 05 h (first extraction) Residuefrom the first extraction was boiled in 8x volume of waterfor 25min Finally the filtered solutions were combined andconcentrated into an aqueous extract containing 12 gmL rawherbs Valsartan was purchased from Novartis Pharma Ltd(Beijing China)

22 Blood Pressure Detection Systolic blood pressure (BP)was monitored prior to treatment (0 weeks) and after 2 4 6and 8 weeks of treatment using a noninvasive computerizedtail-cuff system (BESN-II Desai Production BiotechnologyCo Nanjing China) and performed by specifically assignedinvestigators at regular intervals in order to minimize errorThe mean value from 3 measurements of BP was taken foreach rate representing the sample systolic pressure

23 Pathological Examination of Renal Tissue After 8 weeksof treatment rats were sacrificed under chloral hydrate anes-thesia (35 gkg administered via intraperitoneal injection)Blood was rapidly sampled by abdominal aorta puncture andthe serum was stored at minus80∘C prior to analysis Renal tissuewas excised washed in physiological saline snap-frozen inliquid nitrogen and then stored atminus80∘CKidney tissueswerefixed with 4 paraformaldehyde embedded in paraffin andcut into 3-120583m thick sections Hematoxylin-eosin (HE) stain-ing was performed in order to assess glomerular and vascularinjuries and perivascular lesions such as fibrosis under anelectronic scanner (SIPNOMIC 01579 Zeiss Co Germany)

24 Determination of Urinary Protein over 24 Hours Priorto and after 8 weeks of treatment the rats were placed inmetabolic cages where food was not provided (but water wasavailable) Urine samples were collected over 24 h to calculatethe total urinary output and 5mL was collected for centrifu-gation the supernatant was stored at minus20∘C A commercialradioimmunoassay (RIA) kit (PLA Institute of RIA BeijingChina)was used to estimate urine protein over 24 h in accord-ance with the manufacturerrsquos protocol

25 Determination of Serum Creatinine and Blood UreaNitrogen Levels After 8 weeks of treatment blood wascollected from the abdominal aorta and serum was separatedby centrifugation (1500timesg) at 4∘C for 10min and stored inEppendorf tubes Blood urea nitrogen (BUN) and creatininelevels were determined by the Jaffe and diacetyl-oximemethods in accordance with the manufacturer protocol

26 Determination of Ang II and ADM Content in the Kidneyand Plasma After 8 weeks of treatment rats were sacrificedand blood was sampled from the abdominal aorta Freshkidney tissues were collected weighed quickly ground with




3 Results








4 Discussion



0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast

lowast

(a)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowast lowastlowast

(b)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(c)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(d)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(e)






200120583m

(a)

200120583m

(b)

200120583m

(c)

200120583m

(d)


0

100

200

300

400

500

600

700

800


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowastlowast

lowast

(a)

0100200300400500600700800900

1000


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowast

lowast998779lowast

(b)





0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

Bloo

d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast


(b)


0123456789


Expr

essio

n of

Ang

II in

kid

ney

(pg

g)

lowast

lowast998779lowast

(a)

0

05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

0

50

100

150

200

250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540



Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)





r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















3 Results








4 Discussion



0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast

lowast

(a)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowast lowastlowast

(b)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(c)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(d)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(e)






200120583m

(a)

200120583m

(b)

200120583m

(c)

200120583m

(d)


0

100

200

300

400

500

600

700

800


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowastlowast

lowast

(a)

0100200300400500600700800900

1000


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowast

lowast998779lowast

(b)





0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

Bloo

d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast


(b)


0123456789


Expr

essio

n of

Ang

II in

kid

ney

(pg

g)

lowast

lowast998779lowast

(a)

0

05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

0

50

100

150

200

250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540



Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)





r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowastlowast

lowast

(a)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)

lowast lowastlowast

(b)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(c)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(d)

0

50

100

150

200

250


Bloo

d pr

essu

re o

f rat

s (m

mH

g)


(e)






200120583m

(a)

200120583m

(b)

200120583m

(c)

200120583m

(d)


0

100

200

300

400

500

600

700

800


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowastlowast

lowast

(a)

0100200300400500600700800900

1000


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowast

lowast998779lowast

(b)





0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

Bloo

d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast


(b)


0123456789


Expr

essio

n of

Ang

II in

kid

ney

(pg

g)

lowast

lowast998779lowast

(a)

0

05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

0

50

100

150

200

250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540



Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)





r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















200120583m

(a)

200120583m

(b)

200120583m

(c)

200120583m

(d)


0

100

200

300

400

500

600

700

800


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowastlowast

lowast

(a)

0100200300400500600700800900

1000


The q

uant

ity o

f urin

ary

prot

ein

over

24

h (m

gL)

lowast

lowast998779lowast

(b)





0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

Bloo

d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast


(b)


0123456789


Expr

essio

n of

Ang

II in

kid

ney

(pg

g)

lowast

lowast998779lowast

(a)

0

05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

0

50

100

150

200

250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540



Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)





r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















0102030405060708090


Seru

m cr

eatin

ine l

evel

(120583m

olL

)lowast

lowast998779lowast

(a)

0123456789

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d ur

ea n

itrog

en co

nten

t(m

mol

L)


lowast


(b)


0123456789


Expr

essio

n of

Ang

II in

kid

ney

(pg

g)

lowast

lowast998779lowast

(a)

0

05

1

15

2

25

3


Expr

essio

n of

AD

M in

kid

ney

(pg

g)lowast

lowast998779lowast

(b)

0

50

100

150

200

250


lowast

Con

tent

of A

ng II

in p

lasm

a(p

gm

L)

(c)

05

10152025303540



Con

tent

of A

DM

in b

lood

pla

sma

(pg

mL)

(d)





r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















r = minus0853

P lt 0001

n = 7

0123456789

10


Con

tent

of A

ng II

in k

idne

y (p

gg)

ControlModel

ValsartanJYTL

(a)

r = minus0740

P lt 0001

n = 7

ControlModel

ValsartanJYTL

0

50

100

150

200

250


Bloo

d pr

essu

re (m

mH

g)

(b)









5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















5 Conclusion






Acknowledgments


References































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
































1


















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Documents

Research Article The Renal Protective Effect of Jiangya ...downloads.hindawi.com/journals/ecam/2015/428106.pdfHypertensive nephrosclerosis is one of the most common and serious chronic