| Objective : Based on PI3K/Akt signal pathway,the protective effect of Tiaopihuxin Recipe on myocardial ischemia-reperfusion injury in rats and its possible mechanism were discussed.Methods:(1)Fifty rats were randomly divided into CG group(normal group),S group(sham operation group),I/R group(model group),T+I/R group(Tiaopinhuxinfang group)and Q+I/R group(trimetazidine group),with 10 rats in each group;(2)After 1 week of adaptive feeding,rats in each group were given intragastric administration according to T+I/R group 23.4g crude drug /kg/d;Q+I/R group was given trimetazidine 10mg/kg/d;CG,S and I/R groups were all given the same amount of normal saline.(3)After 3 weeks of intragastric gavage,the MIRI model was made,and the left atrial appendage of the rats was ligated for 30 minutes and then reperfused for 120 minutes.Group S was treated with thread only.After successful modeling,the serum and myocardial tissue of the abdominal aorta of the rats were collected.(4)The pathological tissue of myocardium was observed by HE staining under electron microscope.(5)The contents of CK-MB,AST,cTnI and LDH in serum of rats were detected by ELISA.(6)The contents of PI3 K and Akt protein in myocardial tissue of rats in each group were detected by Western blot.(7)PCR was used to detect the mRNA expression of apoptotic proteins Bax and Bcl-2in the myocardial tissue of experimental rats.(8)SPSS24.0 software was used for statistical analysis of the test data.Results:(1)Pathological manifestations of rat heart:Compared with the normal group,there was no significant change in myocardial tissue in group S,only slight blood stasis and vasodilation.In I/R group,myocardial fibers were broken,twisted and disorderly arranged,intercellular edema,hemorrhage,necrosis,infiltration of inflammatory cells,disordered nuclei,and uneven staining.In the T+I/R group and the Q+I/R group,most of the myocardial cell structures were normal,with few myocardial fiber breakage,a small amount of inflammatory cell infiltration,some nuclei were disordered,and the color was different.(2)Serum contents of CK-MB,AST,cTnI,LDH: CK-MB,cTnI,AST,LDH contents in I/R group were higher than those in S group,and there were differences(P < 0.05).Compared with I/R group,the contents of CK-MB,cTnI,AST and LDH in T+I/R group and Q+I/R group were lower,and there were differences(p < 0.05).The levels of cTnI,CK-MB and LDH in T+I/R group were lower than those in Q+I/R group,and there were differences(P<0.05).(3)Expression of PI3 K and Akt protein: the expression of PI3 K and Akt protein in I/R group was lower than that in S group,and there was a difference(P<0.05);Compared with I/R group,the protein expressions of PI3 K and Akt in T+I/R group and Q+I/R group were significantly increased,and the differences were significant(P<0.05).The protein contents of PI3 K and AKT in Q+I/R group were lower than those in T+I/R group,and there was a difference(P<0.05).(4)mRNA expression of Bcl-2 and Bax: Compared with the control group,Bax gene expression was up-regulated and Bcl-2 gene content and Bcl-2/Bax ratio decreased in ischemia-reperfusion group,with significant difference(P<0.01).Compared with I/R group,Bcl-2 gene expression increased,Bax gene expression decreased and Bcl-2/Bax ratio increased in T+I/R group and Q+I/R group,and the difference was statistically significant(P<0.05).Compared with Q+I/R group,Bcl-2 and Bax gene expression in T+I/R group did not increase significantly(P>0.05),but Bcl-2/Bax ratio increased(P<0.05).Conclusion:(1)Tiaopi Huxin Decoction can obviously reduce the contents of CK-MB,AST,cTnI and LDH in serum of rats,and delay myocardial ischemia-reperfusion injury.(2)Tiaospleen and Huxin formula can improve myocardial pathological changes and protect myocardial cells.(3)Tiaopi Huxin recipe can up-regulate the expression of PI3 K and Akt protein and inhibit apoptosis.(4)Tiaopi Huxin Decoction can reduce the content of Bax mRNA,increase the content of Bcl-2 mRNA and inhibit the apoptosis of myocardial cells.(5)Tiaopinhuxinfang has a protective effect on mi RI,the mechanism of which may be related to the activation of PI3K/ Akt signaling pathway by Tiaopinhuxinfang to regulate apoptosis genes and inhibit apoptosis.(6)The prescription for regulating spleen and protecting heart is safe and effective,and no obvious adverse reactions were observed in animal experiments. |
