Effect and mechanism of all-trans retinoic acid on cyclosporin-induced proliferation and apoptosis of glomerular mesangial cells
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摘要:
目的 探讨全反式维甲酸(ATRA)对环孢素(CsA)诱导的大鼠肾小球系膜细胞增殖及凋亡的影响及机制。 方法 采用不同剂量ATRA作用于不同剂量CsA诱导下的肾小球系膜细胞。采用MTT比色法检测细胞增殖,Hoechst 33258荧光染色观察细胞凋亡形态,后用流式细胞仪检测细胞凋亡率,免疫荧光法检测线粒体促凋亡的Smac蛋白的表达。 结果 与对照组比较,CsA剂量为0.5 μg/mL及以上可抑制细胞増殖,CsA剂量为1.0 μg/mL及以上可诱导细胞凋亡,且Smac蛋白的表达随CsA剂量的增加而增加,具有剂量和时间依赖的特征(均为P<0.05)。而与CsA组比较,CsA+ATRA处理组对细胞增殖的抑制作用更为明显,而加入ATRA可呈剂量依赖性抑制CsA诱导的肾小球系膜细胞凋亡及Smac蛋白的表达(均为P<0.05)。 结论 CsA可抑制肾小球系膜细胞增殖,诱导其凋亡并刺激Smac蛋白的表达。ATRA可以抑制CsA诱导的肾小球系膜细胞凋亡,且可能是通过Smac信号通路介导的。 Abstract:Objective To investigate the effect and mechanism of all-trans retinoic acid (ATRA) on the cyclosporin (CsA)-induced proliferation and apoptosis of glomerular mesangial cells in rat models. Methods The glomerular mesangial cells induced by different doses of CsA were treated with different doses of ATRA. MTT assay was carried out to detect cell proliferation. Hoechst 33258 fluorescent staining was adopted to observe the morphology of the apoptotic cells. Flow cytometry was conducted to detect the cellular apoptosis rate. Immunofluorescent staining was employed to quantitatively measure the expression level of mitochondria-derived pro-apoptotic Smac protein. Results Compared with the control group, administration of CsA at a dose of 0.5 μg/mL and above could suppress cellular proliferation, and use of CsA at a dose of 1.0 μg/mL and above could induce cellular apoptosis. The expression level of Smac protein was significantly up-regulated by CsA administration with a dose and time dependence (all P < 0.05).Compared with the CsA group, combined administration of CsA and ATRA exerted a more significant inhibitory effect on cellular proliferation. Supplement of ATRA could significantly inhibit glomerular mesangial cellular apoptosis induced by CsA and down-regulate the expression of Smac protein with a dose dependence (both P < 0.05). Conclusions CsA can inhibit cellular proliferation, induce cellular apoptosis and up-regulate the expression of Smac protein of glomerular mesangial cells. ATRA is capable of suppressing glomerular mesangial cellular apoptosis induced by CsA, which is probably mediated by the Smac signaling pathway. -
表 1 ATRA和CsA对肾小球系膜细胞增殖的影响
Table 1. Effect of ATRA and CsA on proliferation of glomerular mesangial cell
(x±s) 组别 n A570值 24 h 48 h 72 h A组 3 0.672±0.028 0.708±0.016 0.723±0.035 B组 3 0.635±0.020a 0.592±0.029a,d 0.549±0.018a,d C组 3 0.587±0.031a 0.523±0.024a,d 0.459±0.033a,d D组 3 0.471±0.025a 0.416±0.019a,d 0.362±0.023a,d E组 3 0.418±0.026a 0.365±0.022a,d 0.314±0.030a,d F组 3 0.668±0.024 0.702±0.018 0.718±0.030 G组 3 0.384±0.019a,b,c 0.316±0.028a,b,c,d 0.253±0.025a,b,c,d H组 3 0.313±0.036a,b,c 0.247±0.016a,b,c,d 0.182±0.014a,b,c,d I组 3 0.249±0.027a,b,c 0.175±0.030a,b,c,d 0.116±0.023a,b,c,d 与A组比较,aP<0.01;与E组比较,bP<0.05;与F组比较,cP<0.05;与同组24 h比较,dP<0.05 -
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