| Objective: To observe the effect of the serum containing Modified Xuefu Zhuyu decoction on the SHH path-related proteins(SHH,Ptch 1,Smo,Gli1),angiogenic factors(VEGF,FGF,Ang1)and vascular endothelial markers(CD31,CD34,CD105)of rat cardiac microvascular endothelial cells after oxygen glucose deprivation/reoxygenation(OGD/R),and to explore the mechanism of Modified Xuefu Zhuyu decoction on the OGD/R of rat cardiac microvascular endothelial cells.Methods: Prepare the drug-containing serum of Modified Xuefu Zhuyu decoction and tanshinoneⅡA solution.After the rat heart microvascular endothelial cells were cultured and passaged,they were randomly divided into blank group,model group,traditional Chinese medicine group(Modified Xuefu Zhuyu decoction Formula),traditional Chinese medicine plus inhibitor group,and tanshinone(tanshinone ⅡA)group.Except for the blank group,oxygen glucose deprivation/reoxygenation model was made after pretreatment,and the expression of angiogenesis factors VEGF,FGF,Ang1 was detected by PCR;Elisa detected the expression of vascular endothelial markers CD31,CD34 and CD105;WB detection of SHH signal pathway related proteins SHH,Ptch 1,Smo,Gli1 expression.Rusults:1.The results of CCK-8 showed that :(1)compared with the 5% concentration group of traditional Chinese medicine,the cell viability of the 10% group of traditional Chinese medicine significantly increased at 2h,4h,and 8h(P<0.01),but there was no statistically significant difference at 12h(P>0.05);The cell viability of the 20% group of traditional Chinese medicine significantly decreased at 4 and 12 hours(P<0.01);The comparison at 2 and 8 hours was not statistically significant(P>0.05).Compared with the 10% group of traditional Chinese medicine,the cell viability of the 20% group of traditional Chinese medicine decreased significantly at 2h,4h,8h,and 12h;(2)0.1μmol/L group of tanshinone IIA Compared with the 1μmol/L group,The cell viability of the 1μmol/L group significantly increased at 2h,4h,and 8h(P<0.01),but the comparison at 12 h was not statistically significant(P>0.05);The cell viability of the 10μmol/L group significantly decreased at 2h,4h,and 8h(P<0.01),but the comparison at 12 h was not statistically significant(P>0.05).And compared with the 1μmol/L group,The cell viability of the 10μmol/L group significantly decreased at 2h,4h,8h,and 12h(P<0.01);(3)Through data analysis,it was found that at 4 hours,both intervention methods had higher cell promoting activity.Therefore,in this experiment,a pretreatment time of 4 hours was used,and the concentration of serum containing medicine used in the experiment was 10% Chinese herbal medicine serum and 1μmol/L tanshinone IIA for subsequent experiments.2.The expression of angiogenesis factors VEGF,FGF and Ang1 showed that compared with the blank group,the m RNA expression of VEGF,FGF and Ang1 in the model group was significantly increased(P<0.01,P<0.05);Compared with the model group,the m RNA expression of VEGF,FGF and Ang1 in the traditional Chinese medicine group and tanshinone group were significantly increased(P<0.01);The m RNA expression of VEGF was significantly decreased in the Chinese medicine plus inhibitor group(P<0.01),and the m RNA expression of FGF and Ang1 had a downward trend,but there was no statistical significance(P>0.05);Compared with the traditional Chinese medicine group,the m RNA expression of VEGF,FGF and Ang1 in the traditional Chinese medicine plus inhibitor group decreased significantly(P<0.01),but there was no significant difference compared with the tanshinone group(P>0.05);Compared with the traditional Chinese medicine plus inhibitor group,the m RNA expression of VEGF,FGF and Ang1 in the tanshinone group was significantly increased(P<0.01).3.The expression of vascular endothelial markers CD31,CD34 and CD105 showed that compared with the blank group,the expression of CD31,CD34 and CD105 in the model group increased significantly(P<0.05,P<0.01);Compared with the model group,the expression of CD31,CD34 and CD105 in the traditional Chinese medicine group and tanshinone group were significantly increased(P<0.01);The expression of CD105 was significantly decreased in the Chinese medicine plus inhibitor group(P<0.01),while the expression of CD31 and CD34 was not statistically significant(P>0.05);Compared with the traditional Chinese medicine group,the expression of CD31,CD34 and CD105 in the traditional Chinese medicine plus inhibitor group decreased significantly(P<0.01),and the expression of CD34 and CD105 in the tanshinone group also increased significantly(P<0.05),but the comparison of CD31 was not statistically significant(P>0.05);Compared with the traditional Chinese medicine plus inhibitor group,the expression of CD31,CD34 and CD105 in tanshinone group were significantly increased(P<0.01).4.The expression results of SHH pathway-related proteins SHH,Ptch 1,Smo and Gli1 showed that compared with the blank group,the expression of SHH,Ptch 1,Smo and Gli1 proteins in the model group increased significantly(P<0.01);Compared with the model group,the expression of SHH,Ptch 1,Smo and Gli1 protein in the traditional Chinese medicine group and tanshinone group were significantly increased(P<0.01);The expression of SHH,Ptch 1,Smo and Gli1 protein in the Chinese medicine plus inhibitor group decreased significantly(P<0.01);Compared with the traditional Chinese medicine group,the expression of SHH,Ptch 1,Smo and Gli1 protein in the traditional Chinese medicine plus inhibitor group decreased significantly(P<0.01);Compared with the traditional Chinese medicine plus inhibitor group,the expression of SHH,Ptch 1,Smo and Gli1 protein in the tanshinone group was significantly increased(P<0.01).Conclusion:(1)Modified Xuefu Zhuyu decoction can promote the expression of angiogenesis factors to promote angiogenesis.(2)Modified Xuefu Zhuyu decoction may promote angiogenesis and protect ischemic myocardium by increasing the expression of SHH signal pathway and regulating the expression of vascular endothelial growth factor and vascular endothelial marker. |
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