[1]张姹,张佳幸,熊一凡,等.基于内皮细胞的内皮-间质转化筛选肾康丸活性成分[J].南方医科大学学报,2018,(03):312.
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基于内皮细胞的内皮-间质转化筛选肾康丸活性成分()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2018年03期
页码:
312
栏目:
出版日期:
2018-04-03

文章信息/Info

Title:
Screening active ingredients of Shenkangwan that regulate endothelial-mesenchymal transition of endothelial cells in vitro
作者:
张姹; 张佳幸; 熊一凡; 鲁路; 王明清; 刘思明; 肖炜
关键词:
内皮细胞; 内皮-间质转化; 肾康丸; 毛蕊异黄酮
Keywords:
endothelial cells;  endothelial-mesenchymal transition;  Shenkangwan;  calycosin
摘要:
目的基于ALK5靶点，通过分子对接技术对肾康丸的有效成分进行筛选，并验证各分子在高糖模型中的作用，从而筛选 出肾康丸影响内皮细胞内皮-间质转化活性的有效成分，优化组方配伍。方法对初步筛选的9个分子进行活力测定后选定毛 蕊异黄酮、刺芒柄花苷、豆甾醇3个分子；通过设置正常细胞组、高糖损伤组、高糖损伤组+毛蕊异黄酮、高糖损伤组+刺芒柄花 苷、高糖损伤组+豆甾醇组，并用实时荧光定量PCR检测3个分子的α-SMA，vimentin的mRNA表达水平；用Western blot法检测 SMAD2/3磷酸化情况。结果与正常细胞组活力比较，毛蕊异黄酮、刺芒柄花苷、豆甾醇3个分子细胞毒性弱，与正常细胞组活 力比较差异无统计学意义（P>0.05）。其中，高糖加毛蕊异黄酮组的α-SMA、vimentin的mRNA水平显著下降（P<0.05），SMAD2/ 3 磷酸化受到抑制。其他两组差异无统计学意义。结论高糖对内皮细胞会产生损伤，可通过黄芪单体毛蕊异黄酮来抑制α- SMA、vimentin上调，抑制SMAD2/3参与调节内皮细胞内皮-间质转化从而改善糖尿病肾病。
Abstract:
Objective To screen the effective components of Shenkangwan that regulate endothelial-mesenchymal transition in endothelial cells for optimizing prescription of Shenkangwan. Methods ALK5 was identified as one of the target receptors that regulate endothelial-mesenchymal transition of endothelial cells using molecular docking technique. Nine molecules were screened as the candidate effective components in Shenkangwan, among which calycosin, ononin and stigmasterol were selected for testing. Glomerular epithelial cells were exposed to high glucose and treated with calycosin, ononin, or stigmasterol, and the cellular expressions of α-smooth muscle actin (α-SMA) and vimentin mRNA were detected with realtime fluorescence quantitative PCR. The phosphorylation of SMAD2/3 in the cells was detected using Western blotting. Results Calycosin, ononin and stigmasterol did not produce significant cytotoxicity in glomerular epithelial cells (P>0.05). The cells exposed to high glucose and calycosin treatment showed significantly decreased mRNA levels of α-SMA and vimentin (P< 0.05) and inhibited phosphorylation of SMAD2/3. Ononin and stigmasterol did not produce such effects in the cells. Conclusion In endothelial cells with high glucose-induced injury, calycosin can inhibit the up-regulation of α-SMA and vimentin and inhibit phosphorylation of SMAD2/3 to regulate endothelial-mesenchymal transition and improve diabetic nephropathy.

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更新日期/Last Update: 1900-01-01