天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (10): 1089-1092.doi: 10.11958/j.issn.0253-9896.2015.10.001

• 细胞与分子生物学 •    下一篇

1,25(OH)2D3对人肾小球系膜细胞增殖及 mTOR/p70s6K表达的影响

  

  1. 1新疆石河子大学医学院 （邮编 832000）； 2新疆石河子大学第一附属医院肾病科
  • 收稿日期:2015-04-13 修回日期:2015-05-12 出版日期:2015-10-15 发布日期:2015-10-22

Effects of 1,25(OH)2D3 on proliferation and mTOR/p70s6K expressions of human glomerular mesangial cells

  1. 1 Medical College of Shihezi University, Xinjiang 832000, China; 2 Department of Nephrology, the First Affiliated
    Hospital of Medical College of Shihezi University
  • Received:2015-04-13 Revised:2015-05-12 Published:2015-10-15 Online:2015-10-22

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摘要：  摘要：目的 探讨 1,25-二羟基维生素 D3[1,25(OH)2D3]对人肾小球系膜细胞增殖的影响及其在肾小球系膜细胞调控中对 mTOR/p70s6K 信号通路的作用。方法 体外培养人肾小球系膜细胞， 取传代培养至 3~7 代细胞分为 4组： 正常对照组（加含 5%胎牛血清 DMEM 培养基）， VD 组[加 1,25(OH)2D3 10-8 mol/L]， R 组（加雷帕霉素 5 mg/L）， R+VD 组[加雷帕霉素 5 mg/L 及 1,25(OH)2D3 10-8 mol/L]， 干预 48 h。采用细胞增殖与活性检测试剂盒 CCK-8 检测细胞增殖情况， 流式细胞术检测细胞周期时相分布， 免疫荧光法检测细胞中 mTOR、 p70s6K 的表达情况。结果 正常对照组、 VD 组、 R 组、 R+VD 组：（1） 吸光度值 （A450） 依次降低， 组间多重比较差异均有统计学意义 （均 P < 0.05）； 抑制率（IR） 依次升高。（2） G1期细胞比例依次增加， S 期、 G2/M 期依次减少， 增殖指数 （PI） 依次降低， 除 R 组与 VD 组比较差异无统计学意义外， 其余组间多重比较差异均有统计学意义。（3） 系膜细胞中 mTOR、p70s6K 蛋白表达强度均依次降低， 除 R 组与 VD 组比较差异无统计学意义外， 其余组间多重比较差异均有统计学意义。结论 1,25(OH)2D3可显著抑制人系膜细胞增殖， 且其可能通过抑制 mTOR/p70s6K 信号通路调控肾小球系膜细胞增殖。

关键词: 关键词：肾小球系膜细胞, 细胞增殖, 细胞周期, 核糖体蛋白质 S6 激酶类, 70-kDa, 1,25-二羟基维生素 D3, mTOR/p70s6K 信号通路

Abstract:

Abstract： Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu⁃
man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth⁃
ods The cultured human mesangial cells at passage 3-7 were divided into four groups: control group， VD group (addition of
10-8 mol/L of 1,25-dihydroxyvitamin D3 )， R group (addition of 5 mg/L of rapamycin) and R+VD group（addition of 5 mg/L ra⁃
pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3） . Drug incubation last 48 h. The effect of mesangial cell pro⁃
liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of
mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group
than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group
than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. (2) Cells
in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than
those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in⁃
dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group
than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex⁃
cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera⁃
tion significantly through mTOR/p70s6K signaling pathways.

Key words: Key words：mesangial cells, cell proliferation, cell cycle, ribosomal protein S6 kinases, 70-kDa, 1,25 (OH)2D3, mTOR/p70s6K signaling pathway