不同免疫抑制剂对肾小球系膜细胞增殖的影响

Effects of different immunosuppressive agents on mesangial cell proliferation

目的 探讨不同免疫抑制剂对受损的肾小球系膜细胞增殖的影响.方法 在体外用细胞松弛素B可逆性地损害处于增殖期的大鼠肾小球系膜细胞株（HBZY-1细胞）,然后将细胞分成5组：对照组,仅加入橄榄油10 μl;环孢素A（CsA）组,加入CsA 3μg/ml（2.5 mmol/L）;他克莫司（Tac）组,加入Tac 1μg/ml;吗替麦考酚酯（MMF）组,加入MMF 0.3μg/ml;西罗莫司（SRL）组,加入SRL10 ng/ml.在药物处理后6、12和24 h时,采用专业图像分析软件对各组不同时间点的系膜细胞进行形态观察、计数和增殖情况分析.结果 药物处理后,各组HBZY-1细胞的形态均有所恢复.处理后6 h时,除Tac组HBZY-1细胞数明显高于对照组外,其余各组与对照组之间的差异均无统计学意义（P＞0.05）.12 h时,各组HBZY-1细胞数之间的差异均无统计学意义（P＞0.05）.24 h时,对照组和MMF组之间细胞数的差异无统计学意义（P＞0.05）;而其余各组的细胞数均显著增高,与对照组、MMF组以及与组内其他时点相比较,差异均有统计学意义（P＜0.05）,其中尤以CsA组和Tac组的细胞数量增加最为显著,SRL组增加的幅度低于CsA组和Tac组（P＜0.05）.结论 CsA、Tac、SRL和MMF均可不同程度地减轻受损大鼠肾小球系膜细胞的损伤和促进其恢复,但是其中CsA和Tac明显地促进了细胞的增殖,SRL促细胞增殖作用不明显,而MMF则几乎不引起系膜细胞增殖.

Objective To investigate the effects of different immunosuppressive agents on mesangial cell proliferation through a mesangial cell injury model in vitro. Methods Mesangial cell line （HBZY-1） in period of proliferation was cultured in vitro with cytochalasin B for 2 h, then HBZY-1 cells were divided into 5 groups： blank （control） group, cyclosporine A （CsA） group, Tacrolimus （Tac） group, mycophelonate mofetil （MMF） group and rapamycin （RAPA） group. Subsequently,the number of HBZY-1 cells at different time points was measured by using the professional image analysis software after treatment for 6, 12 and 24 h, respectively. Results Damaged HBZY-1 cells recovered in all groups. At 6 h, the number of HBZY-1 cells in Tac group was significantly more than that in control group （P＜0.05）, but the difference had no significance between the other treatment groups and control group （P＞0. 05）. At 12 h, there was no significant difference in of the number of HBZY-1 cells among the all groups （P＞0. 05）. At 24 h, there was no significant difference in the cell number between MMF and control groups （P＞0. 05）. CsA, Tac and RAPA resulted in HBZY-1 cell proliferation, and the cell number in CsA and Tac groups was significantly more than that in the other groups （P＜0. 05）. As compared with the control group, the cell number in RAPA group was significantly increased （P＜0. 05）. Conclusion CsA, Tac, MMF and RAPA contribute to recovery of damaged HBZY-1 cells, but CsA and Tac result in over-proliferation of HBZY-1 cells. RAPA and MMF can prevent HBZY-1 cells against over-proliferation, and MMF scarcely results in HBZY-1 cell proliferation.