白藜三醇对TGF-β诱导的心肌成纤维细胞增殖及相关微小RNA的影响-东南大学学报医学版

白藜三醇对TGF-β诱导的心肌成纤维细胞增殖及相关微小RNA的影响

单位：徐州医科大学附属医院心内科, 江苏徐州 221002

分类号：R-33

出版年·卷·期（页码）：2019·38·第二期（337-342）

摘要：

目的：研究白藜三醇（resveratrol，RSV）对转化生长因子β（TGF-β）诱导的SD新生大鼠心肌成纤维细胞增殖及相关microRNA表达的影响，探讨RSV在microRNA水平上抑制TGF-β诱导的成纤维细胞增殖的可能机制。方法：采用胰酶和胶原酶双酶法及差速贴壁法分离培养新生大鼠心肌成纤维细胞，传代培养，实验中采用2~3代细胞。实验分为对照组、TGF-β（5 ng·ml^-1）组及TGF-β（5 ng·ml^-1）+RSV （50 μmol·L^-1）组3组。CCK-8法检测RSV对心肌成纤维细胞的最佳药物作用浓度、作用时间；CCK-8法检测对照组、TGF-β组及TGF-β+RSV组各组心肌成纤维细胞增殖情况；实时荧光定量PCR （qPCR）TaqMan探针法检测各组细胞miR-34a、miR-181a、miR-17的差异性表达。结果：与对照组相比，TGF-β组心肌成纤维细胞增殖明显增加（P<0.001），与TGF-β组相比，TGF-β+RSV组心肌成纤维细胞增殖明显减少（P<0.001）；与对照组相比，TGF-β组miR-34a、miR-181a及miR-17显著上调，分别为P<0.01、P<0.001、P<0.001；与TGF-β组相比，TGF-β+RSV组miR-34a、miR-181a及miR-17显著下调，分别为P<0.01、P<0.01、P<0.001。结论：RSV可抑制TGF-β诱导的心肌成纤维细胞的增殖，这一过程可能是通过下调miR-34a、miR-181a、miR-17实现的。

Objective:To study the effects of resveratrol (RSV) on the proliferation of cardiac fibroblasts of neonatal SD rats induced by transforming growth factor β (TGF-β) and related microRNA expression, to explore the possible mechanism of inhibiting the proliferation of cardiac fibroblasts at the level of miRNA. Methods:Isolated culture of newborn rats CFs by means of the double-enzymatic method of pancreatic enzymes and collagenase was conducted, and subculture by using 2-3 generations of cell was done. The experiment rats CFs were divided into three groups:control group, TGF-β (5 ng·ml^-1) group and TGF-β (5 ng·ml^-1)+ RSV (50 μ mol·ml^-1) group. The optimal drug effect concentration and time as well as cell proliferation of each group were detected by CCK-8 method. The differential expressions of miR-34a, miR-181a and miR-17 of each group were detected by real-time fluorescence quantitative PCR(qPCR) TaqMan probe method. Results:Compared with the control group, the proliferation of cardiac fibroblasts in the TGF-β group was significantly increased (P<0.001); compared with the TGF-β group, the proliferation of cardiac fibroblasts in the TGF-β+ RSV group was reduced obviously (P<0.001); compared with the control group, miR-34a, miR-181a and miR-17 of the TGF-β group were remarkably up-regulated(P<0.01, P<0.001, P<0.001, respectively); compared with the TGF-β group, miR-34a,miR-181a and miR-17 of the TGF-β+RSV group were obviously down-regulated(P<0.01, P<0.01, P<0.001, respectively). Conclusion:RSV can inhibit the proliferation of cardiac fibroblasts induced by TGF-β,and it may be achieved by down-regulating miR-34a, miR-181a and miR-17.

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