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目的观察1,25(OH)2D3能否抑制高糖导致的大鼠肾小球系膜细胞(RMC)肥大,并探讨其可能的作用机制。方法将体外培养的RMC分为正常对照组、等渗对照组、高糖组、单纯1,25(OH)2D3组、等渗+1,25(OH)2D3组及高糖+1,25(OH)2D3组。1,25(OH)2D3浓度为10-7mol/L,培养48 h,检测各组细胞总蛋白/细胞数比值,流式细胞仪检测各组前向角光强度(FSC),并采用Western blot检测各组维生素D受体(VDR)、哺乳动物雷帕霉素靶蛋白(mTOR)、核蛋白S6激酶(p70S6K)、延长因子4E结合蛋白(4EBP1)的表达情况。结果高糖+1,25(OH)2D3组与高糖组比较,FSC降低(530.7±15.6 vs.509.0±35.0,P<0.05)、总蛋白/细胞数比值降低(41.5±1.7 vs.49.9±1.1,P<0.05);Western blot半定量灰度分析显示,高糖组较正常对照组VDR下调,mTOR及p70S6K的蛋白水平上调(P<0.05);高糖+1,25(OH)2D3组较高糖组mTOR及p70S6K的蛋白水平均下调(P<0.05)。结论 1,25(OH)2D3可逆转高糖导致的RMC肥大,其机制可能是通过抑制mTOR及其下游信号通路,减少了蛋白质的合成。
Objective To investigate whether 1,25 (OH) 2D3 can inhibit the hypertrophy of rat mesangial cells (RMC) induced by high glucose and to explore its possible mechanism. Methods RMCs cultured in vitro were divided into normal control group, isotonic control group, high glucose group, simple 1,25 (OH) 2D3 group, isotonic +1,25 (OH) 2D3 group and high glucose +1,25 OH) 2D3 group. The concentration of 1,25 (OH) 2D3 was 10-7mol / L and cultured for 48 hours. The total protein / cell ratio of each group was measured. The anterior horn light intensity (FSC) of each group was detected by flow cytometry. Western blot The expression of vitamin D receptor (VDR), mammalian target of rapamycin (mTOR), nuclear protein S6 kinase (p70S6K) and elongation factor 4E binding protein (4EBP1) were detected. Results Compared with high glucose group, FSC decreased (530.7 ± 15.6 vs. 509.0 ± 35.0, P <0.05) and total protein / cell ratio decreased (41.5 ± 1.7 vs.49.9 ± 1.1, P <0.05). Western blot semi-quantitative gray-scale analysis showed that VDR was down-regulated and mTOR and p70S6K were upregulated in high glucose group compared with normal control group (P <0.05) The protein levels of mTOR and p70S6K in higher glucose group were all decreased (P <0.05). Conclusions 1,25 (OH) 2D3 can reverse the hypertrophy of RMC induced by high glucose, which may be due to the inhibition of mTOR and its downstream signaling pathways, reducing the protein synthesis.