| Objective:Observation of the effect of Zishen Huoluo prescription to intervene aldosterone escape on the effect of perindopril on the reversal of hypertensive left ventricular hypertrophy,and discuss its possible molecular mechanism.Methods:16 weeks old spontaneously hypertensive rats(SHR)24,SHR were randomly divided into WKY control group,saline group,perindopril group and Zishen Huoluo prescription group.Perindopril group rats were lavaged with perindopril mixed suspension(1mg/kg)in the morning,given equal volume of saline in the afternoon;Zishen Huoluo prescription group rats were lavaged with perindopril mixed suspension(1mg/kg)in the morning,given Zishen Huoluo prescription(12.7g/kg)in the afternoon;saline group and WKY control group rats were lavaged with equal volume physiological saline twice a day,continuously for 12 weeks.The heart rate(HR),systolic blood pressure(SBP),diastolic blood pressure(DBP)and mean pressure(MBP)were measured with caudal artery manometry method at 0th,4th,8th,12 th weekend after administration respectively;The level of serum aldosterone was detected with ELISA method;Hemodynamics was detected with RM6240 biologic signal acquisition system at 12 th weekend;The concentration of type I procollagen carboxyl end propeptide(PICP)and type I collagen carboxyl terminal cross-linked peptide(ICTP)was detected with ELISA method after blood extraction from the abdominal aorta,calculating the PICP/ICTP value;Left ventricular was isolated and measured the left ventricular mass index(LVMI);The content of Hydroxyproline(Hyp)in myocardial tissue was detected with alkali hydrolysis method;Morphology change of myocardial tissue was explored by HE staining;Interaction of mineralcorticoid receptor(MR)and striatin,MR and Caveolin-1(Cav-1)were detected with the Co-IP;p EGFR and p ERK1/2 m RNA protein in myocardium expression were detected with the Western-blot method.Result:(1)Each time point before and after administration,the SBP?DBP and MBP level of saline group rats were significantly higher than WKY control group(P<0.05);After 4weeks of administration,compared with saline group,the SBP level of perindopril group rats decreased significantly(P<0.05);The SBP?DBP and MBP level of Zishen Huoluo prescription group rats decreased significantly(P<0.05);Compared with perindopril group,the DBP and MBP level of Zishen Huoluo prescription group rats decreased significantly(P<0.05).After 8 weeks of administration,compared with saline group,the SBP,DBP and MBP level of perindopril group and Zishen Huoluo prescription group rats decreased significantly(P<0.05);Compared with perindopril group,the SBP,DBP and MBP level of Zishen Huoluo prescription group rats decreased significantly(P<0.05).After 12 weeks of administration,compared with saline group,the SBP,DBP and MBP level of perindopril group and Zishen Huoluo prescription group rats decreased significantly(P<0.05);Compared with perindopril group,the DBP and MBP level of Zishen Huoluo prescription group rats decreased significantly(P<0.05).(2)Each time point before and after administration,the serum aldosterone level of saline group was higher than WKY control group(P<0.05).Compared with saline group,the serum aldosterone level of perindopril group and Zishen Huoluo prescription group rats decreased significantly at 4th,8th weeks(P<0.05).After 12 weeks of administration,compared with saline group and perindopril group,the serum aldosterone level of Zishen Huoluo prescription group rats decreased significantly(P<0.05).(3)Hemodynamic test shows that the +dp/dtmax of saline group was lower than WKY control group,the LVDP?-dp/dtmax of saline group were higher than WKY control group(P<0.05);Compared with saline group and perindopril group,the +dp/dtmax of Zishen Huoluo prescription group rats increased significantly,the LVDP?-dp/dtmax of Zishen Huoluo prescription group rats decreased significantly(P<0.05).(4)The BW of saline group was lower than WKY control group,the LVMI of saline group were higher than WKY control group(P<0.05);Compared with saline group,the LVM?LVMI of perindopril group and Zishen Huoluo prescription group rats decreased significantly(P<0.05);Compared with perindopril group,the LVMI of Zishen Huoluo prescription group rats decreased significantly(P<0.05).(5)The Hyp?PICP?PICP/ICTP of saline group rats were higher than WKY control group,the ICTP of saline group rats was lower than WKY control group(P<0.05);Compared with saline group,the Hyp ? PICP ? PICP/ICTP of Zishen Huoluo prescription group rats decreased significantly(P<0.05).(6)HE staining of the left ventricular papillary muscle section showed that compared with the WKY control group,the ventricular wall of the normal saline group was thicker,the myocardial cells were loosely arranged and the space was enlarged.The left ventricular myocardial longitudinal HE staining showed that myocardial cells in WKY control group rats arranged in neat,compact,clear structure,uniform size nuclear,cytoplasmic staining uniform;The myocardial cell hypertrophy in the saline group rats twisted disorder,irregular nuclear size,can see a lot of myocardial fiber fracture,myocardial cell gap is obvious.Expand.The above pathological changes were alleviated in each group,and the improvement was most obvious in the group of Zishen Huoluo prescription.(7)Co-IP results shows the co-immunoprecipitation of MR and Cav-1 in the group of Zishen Huoluo prescription was higher than saline group,the co-immunoprecipitation of MR and striatin in the group of Zishen Huoluo prescription was lower than saline group(P<0.05);Compared with perindopril group,the co-immunoprecipitation of MR and Cav-1 in the group of Zishen Huoluo prescription was higher than perindopril group,the co-immunoprecipitation of MR and striatin in the group of Zishen Huoluo prescription was lower than perindopril group(P<0.05).(8)Compared with saline group and perindopril group,the expression of p ERK1/2 and p EGFR protein in the heart tissue group of Zishen Huoluo prescription group decreased significantly(P<0.05).Conclusions:(1)The combination of Zishen Huoluo prescription with perindopril has a better antihypertensive effect,and the effect of hypotension is better than perindopril alone.(2)The combination of Zishen Huoluo prescription with perindopril in reducing LVMI,improving cardiac function and alleviating the myocardium pathological injury of SHR is better than perindopril alone.(3)The combination of Zishen Huoluo prescription with perindopril can reduce the content of Hyp in SHR myocardium,inhibit the deposition of extracellular collagen and improve myocardial fibrosis.(4)Aldosterone escape can occured in the long-term perindopril treatment,Zishen Huoluo prescription can overcomed the effect of aldosterone escape,this may be the main reason for its reversal of left ventricular hypertrophy than the efficacy of perindopril.(5)The intervention of Zishen Huoluo prescription on aldosterone escape may be related to membrane translocation and localization of MR,and regulate its downstream EGFR/ERK signaling pathway. |
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