Effect Of YQWY On Cardiac Function In Rats With Pressure Overload Heart Failure Based On The IL-10/STAT3 Pathway

Objective:Heart failure(HF)is a global health problem,cardiac remodel is an important pathophysiological alteration in heart failure.This study aimed to investigate the effects of YQWY on cardiac function,inflammation,fibrosis and apoptosis in rats with pressure load induced heart failure,and to investigate the possible underlying mechanisms.Methods:We selected 30 male wistar rats,which were adaptively raised for 1 week,and 5 male Wistar rats were randomly selected as the sham group(n=5),only isolated arteries did not cause arterial stenosis during surgery,and were fed a normal diet postoperatively.The remaining rats were prepared pressure load induced heart failure witar rat model by minimally invasive aortic arch constriction(MTAC)to confirm the success of the modeling after 4 weeks.The rats were randomly divided into MTAC group(n=5),low-dose group of YQWY(n=5),and high-dose group of YQWY(n=5).YQWY is prepared by soaking,boiling,decoction of the drugs,and finally concentrating using a Chinese medicine rotary steamer.The low-dose group of YQWY was administered 3.6g/kg/d by gavage,and the high-dose group of YQWY was administered 18 g/kg/d by gavage for 16 weeks.After intervention by YQWY,left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS)were measured by echocardiography to evaluate the effects of YQWY on cardiac function.The rats were sacrificed by taking blood from the abdominal aorta of wistar rats at week 16,and cardiac tissue was left.Collagen I,TGF-?,CTGF protein expression levels were determined by Masson staining,Sirius staining,immunohistochemistry of myocardial tissue,and Western blot was used to evaluate the effects of YQWY on myocardial fibrosis.The protein expression levels of Bax,Bcl-2,Caspase-3 and PARP were detected by TUNEL staining and Western blot to evaluate the effect of YQWY on cardiomyocyte apoptosis.The expression levels of IL-10 and TNF-? in serum and myocardial tissues were measured by ELISA to evaluate the effects of YQWY on inflammatory factors.CD68 expression in myocardial tissue was detected by immunohistochemistry,and IL-10,TNF-?,STAT3(p-STAT3),p65(p-p65)expression levels were detected by Western blot to evaluate the effects of YQWY on inflammatory pathways.Results:(1)After preparation of a rat model of pressure load induced heart failure by MTAC,echocardiography revealed the systolic function of Wistar rats was significantly attenuated after modeling,and the left ventricular ejection fraction was decreased,with statistically significant differences(P<0.05),which suggested that the modeling was successful.(2)LVFS and LVEF decreased in the MTAC group compared with the sham group(P<0.01).After intervention with YQWY for 16 weeks,compared with the MTAC group,LVEF increased in the low-dose and high-dose groups,and the differences were statistically significant(P<0.01).LVFS increased in the low-dose group,and the differences were statistically significant(P<0.05).LVFS increased in the high-dose group,and the differences were statistically significant(P<0.01).(3)After Masson and Sirius staining,the percentage of myocardial fibrosis was significantly higher in the MTAC group compared with the sham group,and the difference was statistically significant(P<0.01).After intervention by YQWY,the percentage of myocardial fibrosis area in the low and high dose groups of YQWY was significantly reduced,and the differences were statistically significant(P<0.05).Immunohistochemical detection of myocardial tissue showed that collagen I,TGF-?,and CTGF expression increased in the MTAC group compared with the sham group,whereas after intervention by YQWY,collagen I,TGF-?,and CTGF expression decreased significantly in the low and high dose groups.Myocardial tissue was detected by Western blot,and the protein expression of collagen I,TGF-?,and CTGF in the MTAC group was higher than that in the sham group,whereas after intervention by YQWY,collagen I,TGF-?,and CTGF were significantly downregulated in the low and high dose groups.(4)By TUNEL staining,cardiomyocyte apoptosis was obvious in the MTAC group,whereas it was inhibited in both the low and high dose groups by YQWY.After Western blot assay,YQWY decreased the expression of apoptotic signaling proteins Bax,Caspase-3 and PARP,and increased the expression of Bcl-2 in the myocardium of MTAC rats.(5)After the serum IL-10 and TNF-? expression levels were measured by ELISA,compared with the sham group,the levels of IL-10 in the MTAC group were significantly decreased,TNF-? was significantly increased,and the differences were statistically significant(P<0.01).Compared with the MTAC group,TNF-? was significantly decreased in the low-dose group of YQWY,and the differences were statistically significant(P<0.05).Compared with the MTAC group,the level of IL-10 incresed and TNF-? decreased in the high-dose group of YQWY,the differences were statistically significant(P<0.01).After the myocardial tissue of IL-10 and TNF-? expression levels were measured by ELISA,compared with the sham group,the level of IL-10 in the MTAC group were significantly decreased,the level of TNF-? was significantly increased,the differences were statistically significant(P<0.01),Compared with the MTAC group,the level of IL-10 was significantly increased in the high-dose group of YQWY,the differences were statistically significant(P<0.01).The level of TNF-? was significantly decreased in the high-dose group of YQWY,the differences were statistically significant(P<0.05).(6)CD68 expression was detected in myocardial tissue,which was significantly increased in the MTAC group compared with the sham group,and significantly decreased in the low and high dose groups of YQWY compared with the MTAC group.The inflammatory pathways related to myocardial tissue were detected by Western blot,and the protein expression of IL-10 and STAT3 in myocardial tissue was significantly decreased,and the protein expression of p65 and TNF-?was significantly increased in the MTAC group compared with the sham group,while the protein expression of IL-10 and STAT3 was significantly increased,and the protein expression of p65 and TNF-? was significantly decreased in the low-dose group and high-dose group of YQWY.Conclusions:YQWY reduces myocardial inflammation,fibrosis,and apoptosis,inhibits ventricular remodeling,and protects cardiac function in rats with pressure load induced heart failure.The mechanism may be related to the activation of the IL-10/STAT3 signaling pathway,which improves myocardial remodeling.