山茱萸环烯醚萜总苷对AGEs培养肾小球系膜细胞的影响

目的观察山茱萸环烯醚萜总苷(ICO)对糖化终产物(AGEs)培养肾小球系膜细胞(GMC)形态学、细胞周期及氧化应激的影响。方法用含AGEs的培养液配制ICO、氨基胍,同时设对照。GMC培养48h后,加混合荧光染液进行染色,并立即进行荧光显微观察。另用流式细胞仪分析GMC的周期。试剂盒测定细胞上清液MDA含量与SOD和GSH-Px活性,流式细胞仪检测细胞内ROS含量。结果形态学显示,正常GMC形态结构正常。而GMC在AGEs的刺激下,细胞数量明显增多,细胞结构不甚清晰,细胞S期延长,G0/G1缩短,细胞内SOD、GSH-Px下调,ROS、MDA上调。在系统中加入ICO后细胞异常增殖明显减少,细胞形态趋于正常,使S期细胞减少,SOD、GSH-Px活力提高,ROS、MDA含量降低。结论ICO能对抗AGEs对GMC的损伤作用。其可能是通过抑制氧化应激来保护GMC免受AGEs的损害,从而达到防治糖尿病肾病的目的。

Influence of iridosides of cornus officinalis on glomerular mesangial cells cultured with AGEs

Aim To investigate the influence of iridosides of cornus officinalis(ICO) on morphology, cell cycle and oxidative stress of glomerular mesangial cells(GMC) cultured with advanced glycation end products(AGEs). Methods GMC were incubated in culture medium containing AGEs in the presence of ICO and aminoguanidin for 48 hours. At the same time, the control and model groups were added. Then the cultured GMC were stained by mixed fluorescence liquid and observed under fluorescence microscope. Cell cycle of GMC were analyzed using flowcytometry. The content of MDA, activity of SOD and GSH-Px in GMC supernatant were measured. The level of ROS was detected using flowcytometry. Result Morphology analysis showed that the morphology and structure of normal GMC were normal. The structure of most cells in AGEs were unclear, cell counts increased largely and they grow intensively. Cell cycle analysis showed that cell percentage of S phase increased and G_0/G_1 reduced. The level of ROS, MDA remarkably increased, and SOD, GSH-Px activity reduced. When the ICO were added, cell morphology tended to be basically normal and cell counts decreased, the percentale of S phase also decreased. The level of ROS, MDA and the SOD, GSH-Px activity restored in comparison with the model groups. Conclusion ICO can prevent the cultured GMC from lesion caused by AGEs. ICO may protect GMC from AGES to retard the progression of diabetic nepropathy by partially inhibiting the occurrence of oxidative stress.