The Effect And Possible Mechanisms Of HY Against Myocardial Fibrosis In Spontaneously Hypertensive Rats

Background&ObjectiveHypertension is the primary cause for of myocardial fibrosis and causes a sustained increase in cardiac pressure overload,triggering excessive proliferation and the accumulation of interstitial and perivascular collagen fibers that leads to myocardial fibrosis.Traditional Chinese medicine is the main method to cure diseses for thousands of years.Through its multi-component,multi-path and multi-target synergy,it exerts the therapeutic effect that can not be obtained by western medicine and single Chinese medicine.Our previous experimental studies show that the effect of XFZY on hypertensive myocardial fibrosis is clear,and it can inhibit hypertension myocardial fibrosis through different ways.However,Xue-Fu-Zhu-Yu Decoction(XFZY)consists of 11 Chinese herbal medicines with complex ingredients,which is not conducive to further pharmacological research and mechanism research.We use the theory of traditional Chinese medicine and network pharmacology to reduce and improve the curative effect of XFZY and look for the efficient drugs combination,then using the modern biotechnology means to explain the mechanism.Content&MethodsAccording to traditional Chinese medicine theory,XFZY can be divided into four groups:Huayu(HY)group,Xingqi(XQ)group,Yangxue(YX)group and Tiaohe(TH)group.BATMAN-TCM was used to construct the four groups network of "active ingredients-targets-diseases".The pharmacological analysis was conducted to predict that HY might be an effective drug combination of XFZY.The high-performance liquid chromatographie fingerprint was developed for ideneification of the components of HY and XFZY.12-week-old male SHR were randomized into six groups with five rats in each group:model group(SHR),low-dose HY group(HY-L),middle-dose HY group(HY-M),high-dose HY group(HY-H),XFZY group(XFZY)and captopril group(captopril),WKY as a normal control group(WKY).Rats were administered for 12 weeks.The diastolic and systolic cardiac function was observed by Echocardiography and Non-invasive tail pressure gauge.Cardiomyocyte morphology and cardiac fibrosis were assessed by HE staining,Masson staining and transmission electron microscopy.Biochemical kit was used to to evaluate heart,liver,and renal function relating indicators.The serum drug chemical analysis method of traditional Chinese medicine was used to identify the blood components of HY.The heart tissue in SHR and HY-H group were sequencing differential genes and conduct bioinformatics analysis.The expression of p-PI3k,PI3k,p-AKT,AKT,p-Erk1/2,Erk1/2,p-p38 and p-p38 protein expression were observed by Western bot.ResultsCombination the network pharmacology and traditional Chinese medicine theory,suggesting that Hua Yu group of drugs is the efficient drugs combination and medicinal ingredients in XFZY.HY treatment significantly increased EF and FS(P<0.001),and decreased SBP,DBP and MBP(P<0.05),These results indicated that HY attenuated this cardiac dysfunction.HY decreased CK,CK-MB,ALT,AST,Cr and the urine of UCr(P<0.05).Also,HY decreased heart weight/body weight and left ventricular weight/body weight(P<0.05).HE staining,Masson staining and transmission electron microscopy result that HY treatment reduced inflammatory infiltration and protected the myocardium from fibrotic deposition.HY reduced the protein and mRNA expression of collagen ?,collagen ?,?-SMA,CAT and GPx.HY increased the activities of CAT,GPx,T-NOS,cNOS,and NO,but decreased inducible iNOS and MDA values(P<0.05).Furthermore,HY treatment increased p-PI3k and p-AKT proteins and decreased p-p38 and p-Erkl/2 proteins.ConclusionHY was an effective drug combination of XFZY that exerted anti-myocardial fibrosis effect,and there was not statistically different between HY and XFZY.HY can significantly reduce blood pressure in SHR rats,also HY can improve cardiac function and protect on myocardial injury,and it has no toxic reactions of liver an kidney.HY could activate PI3k/AKT and inhibite MAPK signaling pathways.Moreover,HY can could increase the antioxidant enzymes and reduce oxidative stress.