甘草酸对AGEs培养肾小球系膜细胞及ECM表达的影响

Effects of glycyrrhizic acid on glomerular mesangial cells and ECM expression induced by AGEs

目的观察甘草酸(glycyrrhizic acid,GA)对糖化终产物(advanced glycation endproducts,AGEs)诱导的大鼠肾小球系膜细胞(HBZY-1)细胞增殖、细胞周期及细胞外基质成分中纤粘连蛋白(FN)和Ⅳ型胶原(C-Ⅳ)的影响。方法用含AGEs的培养液配制GA,同时设牛血清白蛋白(BSA)对照组。HBZY-1各组细胞培养48 h后,MTT检测细胞增殖,流式细胞仪分析细胞周期,ELISA方法测定细胞上清液中FN和C-Ⅳ。结果在AGEs的刺激下,细胞增殖明显,细胞S期延长,G1期缩短,PI从35.01%±4.21%增长到44.93%±0.25%(P<0.05),细胞分泌FN和C-Ⅳ明显增加(P<0.05)。加入GA后,细胞异常增殖明显减少,S期细胞减少,G1期增加,PI从44.93%±0.25%减少到42.16%±1.04%(P<0.05),细胞分泌FN和C-Ⅳ有所减少(P<0.05)。结论 GA能对抗AGEs对HBZY-1细胞的损伤作用,其可能是通过抑制AGEs诱导细胞的异常增殖和改变细胞周期来保护细胞免受AGEs的损害,另外,甘草酸能降低ECM中FN和C-Ⅳ的合成。

Aim To investigate the influence of glycyrrhizic acid（GA） on cell proliferation and cell cycle and secretion of extracellular matrix （ ECM）, the level of fibronection （FN）, type IV collagen （C-IV） in the rats＇ glomerular mesangial cells （HBZY-1） cultured with advanced glycation endproducts （AGEs）. Methads HBZY-1 were incubated in culture medium containing AGEs in the presence of GA and treated for 48 h. At the same time, the normal and control groups were established. Methylthiazoletetrazolium （Mr/T） assay was used to measure cell proliferation; cell cycle analysis was performed using a flow cytometry; FN and C-IV were analyzed by enzyme-linked immunosorbent assay （ELISA）. Results Cell counts increased markedly in AGEs group. Cell eyele analysis showed that cell percentage of S phase increased, G1 reduced and PI increased from 35.01% ±4. 21% to 44. 93% ±0. 25% （P 〈 0. 05）. FN and C-IV secretion increased （P 〈 0. 05 ）. Whereas GA was added, cell counts decreased, and the percentage of S phase also decreased and G1 increased, PI decreased from 44.93% ±0.25% to42.16% ±1.04% （P〈0.05）. In addition, the secretion of FN and C-IV decreased （P 〈 0. 05 ）. Conclusion GA could prevent AGEs- induced HBZY-1. GA may protect HBZY-1 from inhibiting the abnormal cell proliferation, changing the cell cycle. GA can reduce the synthesis of FN and C- IV.