丹酚酸A对心肌成纤维细胞在高浓度葡萄糖刺激下Wnt信号通路的影响

韩路拓; 关瑜; 任钧国; 刘建勋; 郭宏伟; 杨佳妹; 王攀

doi:10.13898/j.cnki.issn.1000-2200.2022.01.005

丹酚酸A对心肌成纤维细胞在高浓度葡萄糖刺激下Wnt信号通路的影响

Effects of salvianolic acid A on Wnt signaling pathway in cardiac fibroblasts stimulated by high concentration glucose

摘要

摘要:
目的探讨丹酚酸A对高浓度葡萄糖诱导的心肌成纤维细胞（CFs）Wnt信号通路的影响及作用机制。
方法采用高浓度葡萄糖刺激CFs构建细胞增殖模型，以22 mg/L卡托普利为西药对照，不同浓度丹酚酸A处理CFs。MTT比色法检测细胞的增殖能力，流式细胞术分析细胞增殖周期，ELISA法测定细胞中Ⅰ型、Ⅲ型胶原的生成和转化生长因子-β1（TGF-β1）蛋白的分泌，蛋白质印迹法检测β-catenin、GSK-3β、p-GSK-3β蛋白的表达水平。
结果干预24、48 h，与模型对照组相比，25、50、100 mg/L丹酚酸A均可明显抑制细胞增殖（P < 0.01）。与模型对照组相比，25、50、100 mg/L丹酚酸A均能抑制CFs Ⅰ型胶原和Ⅲ型胶原的分泌（P < 0.05~P < 0.01）；50、100 mg/L丹酚酸A均能明显抑制TGF-β1的合成及β-catenin的表达（P < 0.01）；100 mg/L丹酚酸A可以上调p-GSK-3β的表达（P < 0.05）。
结论丹酚酸A可抑制高浓度葡萄糖诱导的CFs增殖、胶原蛋白分泌和TGF-β1的合成，其机制可能与抑制Wnt信号通路有关。

Abstract:
ObjectiveTo investigate the effect and mechanism of salvianolic acid A on Wnt signaling pathway in cardiac fibroblasts(CFs) induced by high concentration glucose.
MethodsCell proliferation model was established by stimulating CFs with high concentration glucose, 22 mg/L captopril was used as western medicine control, and CFs were treated with different concentrations of salvianolic acid A.MTT assay was used to detect the proliferation ability of cells, flow cytometry was applied to analyze the cell proliferation cycle, and ELISA was employed to determine the production of type Ⅰ collagen, type Ⅲ collagen and protein secretion of transforming growth factor-β1(TGF-β1), and Western blotting was performed to detect the protein expression level of β-catenin, GSK-3β and p-GSK-3β.
ResultsAfter treatment for 24 and 48 hours, compared with the model control group, 25, 50 and 100 mg/L salvianolic acid A could significantly inhibit cell proliferation(P < 0.01).Compared with the model control group, 25, 50 and 100 mg/L salvianolic acid A inhibited the secretion of type Ⅰ collagen and type Ⅲ collagen in CFs(P < 0.05 to P < 0.01);50 and 100 mg/L salvianolic acid A significantly inhibited the TGF-β1 production and β-catenin expression(P < 0.01);100 mg/L salvianolic acid A upregulated p-GSK-3β expression(P < 0.05).
ConclusionsSalvianolic acid A can inhibit the proliferation, collagen secretion and TGF-β1 production of CFs induced by high concentration glucose, the mechanism of which may be related to the inhibition of Wnt signaling pathway.

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