| Objective:Through the primary culture of rat bone marrow endogenous progenitor cells(Endothelial Progenitor Cells,EPCs),Angiotensin Ⅱ(AngⅡ)induced EPCs senescence,study the effects of Xuefuzhuyu decoction on the senescence,migration and the expression of miR-34 a,p53 and SIRT1 of bone marrow-derived EPCs,and to explore the effect and mechanism of Xuefuzhuyu Decoction on aging of bone marrow-derived EPCs in rats,and to provide scientific basis for the clinical application of Xuefuzhuyu Decoction.Methods:Primary cultured rat bone marrow-derived EPCs and AngⅡ induced EPCs senescence,randomly divided into normal group,model group,miR-34 a inhibitor group,5% Xuefuzhuyu decoction drug-containing serum group(hereinafter referred to as 5% drug-containing serum group),10% Xuefuzhuyu Decoction drug-containing serum group(hereinafter referred to as 10% serum containing serum group),15%Xuefu Zhuyu Decoction drug-containing serum group(hereinafter referred to as 15%drug serum group),cell aging kit detects EPCs aging,Transwell chamber detects EPCs migration function,and the expression of miR-34 a,p53 mRNA and SIRT1 mRNA in EPCs were detected by qRT-PCR.The expression of p53 protein and SIRT1 protein in EPCs were detected by western blot.Results:1.Effects of on EPCs Aging: Compared with the normal group,the number of aging EPCs in model group increased significantly(p<0.01).Compared with the model group,the number of aging EPCs in the miR-34 a inhibitor group and 5%,10%and 15% drug-containing serum group was significantly decreased(p<0.01).Compared with the miR-34 a inhibitor group,the number of aging EPCs in 5%,10%and 15% serum containing sera was significantly increased(p<0.01).Compared with the 5% serum group,the number of aging EPCs in 10% and 15% serum containing serum was significantly decreased(p<0.01).Compared with the 10% serum group,the number of aging EPCs in 15% serum containing serum was significantlydecreased(p<0.01).The concentration of EPCs was the best in 15% of the three serum concentrations,and there was no significant difference in the time of intervention(p>0.05).2.Effect of EPCs migration: Compared with the normal group,the number of EPCs migrated in the model group was significantly decreased(p<0.01).Compared with the model group,the number of EPCs migrated in the miR-34 a inhibitor group and 5%,10% and 15% drug-containing serum group was significantly increased(p<0.01).Compared with the miR-34 a inhibitor group,the number of migrated EPCs in5%,10% and 15% serum containing serum was significantly decreased(p<0.01).Compared with the 5% serum group,the number of EPCs migrated in 10% and 15%serum containing serum was significantly increased(p<0.01).Compared with the10% serum group,the number of EPCs in the 15% drug-containing serum group was significantly increased(p<0.01).Among the three concentrations,15% of the serum containing the group was the best.3.Effects on miR-34a: Compared with the normal group,the expression of miR-34 a in the model group was significantly increased(p<0.01).Compared with the model group,the expression of miR-34 a in the miR-34 a inhibitor group and 10% and15% serum containing serum was significantly decreased(p<0.01),there was no significant difference in the expression of miR-34 a between the 5% serum group and the model group(p>0.05).Compared with miR-34 a inhibitor group,the expression of miR-34 a was significantly increased in 5%,10% and 15% serum(p<0.01).Compared with the 5% serum group,the expression of miR-34 a in 10% and 15% serum containing serum was significantly decreased(p<0.01);Compared with the 10%serum group,the expression of miR-34 a in the serum containing 15% was not statistically significant(p>0.05).The concentration of EPCs was the best in 15% of the three serum concentrations,and there was no significant difference in the time of intervention(p>0.05).4.Effects on p53: Compared with the normal group,the expression of p53 mRNA and p53 protein in the model group was significantly higher than that in the normal group(p<0.01).Compared with the model group,the expression of p53 mRNA and p53 protein in miR-34 a inhibitor group and 10% and 15% containing serum was significantly decreased(p<0.01),the expression of p53 mRNA and p53 protein in 5% serum group was not significantly different from that in model group(p>0.05).Compared with miR-34 a inhibitor group,the expression of p53 mRNA and p53 protein in 5%,10% and 15% serum samples were significantly increased(p<0.01).Compared with the 5% serum group,the expression of p53 mRNA and p53 protein in 10% and 15% serum samples were significantly decreased(p<0.01);Compared with the 10% serum group,the expression of p53 mRNA and p53 protein in the serum containing 15% was not significantly different(p>0.05).The expression of p53 mRNA and p53 protein in 15% serum samples were the best,and there was no significant difference at different time points(p>0.05).5.Effects on SIRT1:Compared with the normal group,the expression of SIRT1 mRNA and SIRT1 protein in the model group was significantly decreased(p<0.01).Compared with the model group,the expression of SIRT1 mRNA and SIRT1 protein in the miR-34 a inhibitor group and 10% and 15% drug-containing serum group were up-regulated(p<0.01),there was no significant difference in the expression of SIRT1 mRNA and SIRT1 protein in 5% serum(p>0.05).Compared with the miR-34 a inhibitor group,the expression of SIRT1 mRNA and SIRT1 protein in 5%,10% and 15% serum samples were significantly decreased(p<0.01);Compared with the 5% serum group,the expression of SIRT1 mRNA and SIRT1 protein in 10% and15% serum samples were significantly different(p<0.01).Compared with the 10%serum group,the expression of SIRT1 mRNA and SIRT1 protein in 15% serum containing sera were significantly increased(p<0.01).Three concentrations of serum containing 15% of the best serum,there was no significant difference in the expression of SIRT1 mRNA at different time points(p>0.05).The effect of SIRT1 protein on 48 h was the best.Conclusions:1.Xuefu Zhuyu Decoction can delay the aging of endothelial progenitor cells and enhance the migration ability of endothelial progenitor cells.2.Xuefuzhuyu Decoction can inhibit the expression of miR-34 a in senescentEPCs.Among them,15% serum containing serum was the best.3.Xuefuzhuyu decoction can down-regulate the expression of p53 in senescent EPCs,among which 15% drug-containing serum group had the best effect.4.Xuefuzhuyu Decoction can up-regulate the expression of SIRT1 in senescent EPCs,among which 15% drug-containing serum group had the best effect... |
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