摘要
目的探讨雷帕霉素对高糖诱导肾小球系膜细胞(GMC)增殖的影响及其可能涉及的途径。方法高糖培养GMC,以不同剂量雷帕霉素干预,MTT及流式细胞仪等方法观察雷帕霉素对细胞增殖和周期的影响,RT-PCR、Western印迹观察细胞Cyclin D1、CyclinE和p27^(KIP1)的变化。结果高糖刺激下,GMC增殖明显;雷帕霉素能抑制这一作用,且呈剂量依赖性,并下调Cyclin D1、Cyclin E的基因及蛋白水平,上调p27^(KIP1)的蛋白表达;与对照组相比,高糖组G_0/G_1期细胞减少,S期细胞比例增加(P均<0.05);雷帕霉素干预后,G_0/G_1期细胞升高,S期细胞比例减少(P均<0.05)。结论雷帕霉素可抑制高糖状态下GMC的增殖,且呈剂量依赖性;其可能机制是通过下调Cyclin D1、Cyclin E及上调p27^(KIP1),参与了G_1/S期阻滞。
Objective To study the effect of rapamycin on cell proliferation of glomerular mesangial cells （GMC） induced by high glucose. Methods Glomerular mesangial ceils were cultured with high glucose and different concentration of rapamycin. Cell proliferation rate was measured by MTT. The mRNA and protein levels of cyclin D1, cyclin E and p27^KIP1 were examined by reverse transcription PCR （RT-PCR） and Western blot analysis respectively. The changes in cell cycles were detected by flow cytometery. Results High glucose could induce GMC prolification, and rapamycin could significantly and dose-dependently inhibit the effect （P〈 0.05）. The mRNA and protein expressions of cyclin D1 and cyclin E were more down-regulated in rapamycin groups than in high glucose group. And the p27^KIP1 protein expression was more up-regulated in rapamycin groups than in high glucose group. In high glucose versus control groups, cell cycle distribution of G0/G1 stage were much lower （61.8 % ± 0. 002 % versus 83. 8 % ± 0.017 % ） and S stage much higher （ 26. 2 % ± 0. 022 % versus 0. 2% ± 0. 004%） （P〈 0. 05）. Under cultured cells with 1,2,5ng/ml of rapamycin, the cell cycle distribution of G0/G1 stage became much higher （61.8% ± 0. 017% versus 63. 3% ±0. 034%, 69. 5% ±0.013%, 73.3%± 0.001% respectively, and S stage much lower （26.2%± 0.022% versus 19.0~ 4- 0. 027%, 12. 0%-±0. 016%, 9： 6%±0. 009%） （P〈0. 05）. Conclusions Rapamycin could dose-dependently inhibit prolification of glomerular mesangial cells induced by high glucose. The possible mechanism is that rapamycin participates the G1/S arrest by down-regulating the mRNA and protein expressions of cyclin D1 and cyclin E, and upregulating p27^KIP1 protein level.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2009年第2期139-142,共4页
Chinese Journal of Diabetes
