扶正化瘀方对血管紧张素Ⅱ诱导的心肌成纤维细胞miR-29b-5p表达的影响

Effect of Fuzheng Huayu Recipe on Expression of miR-29b-5p in Cardiac Fibroblasts Induced by Angiotensin Ⅱ

目的观察扶正化瘀方(FZHY)对血管紧张素Ⅱ(AngⅡ)诱导的心肌成纤维细胞(CFs)增殖、凋亡和miR-29b-5p表达的影响,探讨其抗心肌纤维化的可能机制。方法 差速贴壁法提取乳鼠CFs,以细胞计数试剂盒8法检测细胞的增殖情况;以流式细胞术检测细胞的凋亡情况;以实时荧光定量聚合酶链反应法检测miR-29b-5p的表达情况。结果 与空白对照组相比,AngⅡ组CFs的增殖明显增多(0.519±0.058比0.510±0.020)(P<0.01);与AngⅡ组相比,25、50、100 μg/mL FZHY组CFs增殖明显减少(0.541±0.032,0.529±0.016,0.498 ±0.026比 0.575 ±0.024)(P<0.05或P<0.01),且呈剂量依赖性。与空白对照组相比,AngⅡ组的早期凋亡率和总凋亡率明显减少[(13.017±2.277)%比(19.783±3.082)%、(17.500±2.673)%比(26.067±2.947)%](P<0.01);与AngⅡ组相比,FZHY组早期凋亡率和总凋亡率增加[(19.175±3.384)%比(13.017±2.277)%、(24.875±2.649 )%比(17.500±2.673 )%](P<0.01)。与空白对照组相比,AngⅡ组miR-29b-5p的表达降低(0.729±0.119比1.017±0.218)(P<0.05);与AngⅡ组相比,FZHY组miR-29b-5p升高(1.033±0.235比0.729±0.119)(P< 0.05)。结论 扶正化瘀方抗心肌纤维化的作用机制可能与其抑制AngⅡ诱导的CFs增殖,并促进CFs的凋亡及miR-29b-5p表达有关。

Objective To observe the effect of Fuzheng Huayu recipe(FZHY) on proliferation,apoptosis and expression of miR-29b-5p in cardiac fibroblasts(CFs) induced by angiotensinⅡ(AngⅡ) and to explore the possible mechanism of FZHY in treating myocardial fibrosis.Methods Neonatal rat cardiac fibroblasts were isolated from rats by differential adhesion method.The proliferation of cells was detected by cell counting kit-8.The apoptosis of cells was detected by flow cytometry.The expression of miR-29b-5p was detected by real-time fluorescence quantitative polymerase chain reaction.Results Compared with blank control group,the proliferation of CFs in AngⅡ group was increased significantly(0.519±0.058 vs 0.510±0.020)(P<0.01).Compared with AngⅡ group,the proliferation of CFs in 25,50 and 100 μg/mL FZHY group was decreased significantly(0.541±0.032,0.529±0.016,0.498±0.026 vs 0.575±0.024)(P<0.05 orP<0.01),in a dose dependent manner.Compared with blank control group,the early apoptotic rate and total apoptotic rate in AngⅡ group were decreased significantly[(13.017±2.277)% vs (19.783±3.082)%,(17.500 ±2.673)% vs (26.067 ±2.947)%](P<0.01).Compared with AngⅡ group,the early apoptotic rate and total apoptotic rate of FZHY group were increased[(19.175±3.384)% vs (13.017±2.277)%,(24.875±2.649)% vs (17.500±2.673)%](P<0.01).Compared with blank control group,the expression of miR-29b-5p in AngⅡ group was decreased(0.729±0.119 vs 1.017±0.218)(P<0.05).Compared with AngⅡ group,the expression of miR-29b-5p in FZHY group was increased (1.033±0.235 vs 0.729±0.119)(P<0.05).Conclusion The mechanism of FZHY on myocardial fibrosis may be related to its inhibition of CFs proliferation induced by Ang Ⅱ and promoting the apoptosis of CFs and the expression of miR-29b-5p.