摘要
目的探讨骨形态发生蛋白-7(BMP-7)对高糖诱导系膜细胞增生的作用及其可能的机制。方法体外培养大鼠肾小球系膜细胞,将细胞分为正常对照组(5.5 mmol.L-1葡萄糖),高糖组(25 mmol.L-1),100 ng.L-1 BMP-7组,高糖+100 ng.L-1BMP-7组。应用细胞计数、MTT增殖实验及流式细胞术测定系膜细胞增生和细胞周期变化;应用Real-Time RT-PCR、West-ern blot检测细胞周期蛋白Cyclin D1和Cyclin A2的表达;应用荧光探针2,7-二氯二氢荧光素乙酰乙酸(DCFDA)检测细胞内活性氧(ROS)的变化。结果 (1)BMP-7抑制高糖诱导的系膜细胞增生;(2)高糖诱导系膜细胞从G0/G1期进入S期,而BMP-7能够阻断该作用;(3)BMP-7抑制高糖诱导的周期蛋白Cyclin D1和Cyclin A2核酸及蛋白质表达;(4)BMP-7抑制高糖诱导的活性氧生成。结论 BMP-7抑制高糖诱导的活性氧生成,抑制周期蛋白的表达,进而抑制高糖诱导系膜细胞增生。
Aim To explore the effects of born morphogenetic protein-7（BMP-7）on high glucose-induced rat glomerular mesangial cells（MC）proliferation and its possible mechanisms in vitro.Methods Cultured rat mesangial cells were divided into four groups：normal group with 5.5 mmol·L-1 glucose（NG）,high glucose group with 25 mmol·L-1 D-glucose（HG）,100 ng·L-1BMP-7 and HG ＋100 ng·L-1BMP-7 groups.MC proliferation was measured by cell counting and MTT assay.MC cell-cycle was analyzed by flow cytometry.The levels of cyclinD1 and cyclin A2 were determined by using Real-Time PCR and Western blotting analysis.Reactive oxygen species was assessed by 2,7-dichlorofluorescein diacetate（DCFDA）fluorescence.Results BMP-7 inhibited the HG-induced MC proliferation.HG treatment decreased the G0/G1 phase cells,and increased the S phase cells.BMP-7 inhibited the effect of HG on MC cell cycle progression.BMP-7 inhibited HG-induced expressions of cyclin D1 and cyclin A2.BMP-7 inhibited HG-induced accumulation of ROS in MCs.Conclusion BMP-7 may inhibit HG-induced MC proliferation via blocking the generation of ROS,and it may be one of the potential therapeutic targets against diabetic nephropathy.
出处
《安徽医药》
CAS
2011年第6期685-687,共3页
Anhui Medical and Pharmaceutical Journal
关键词
高糖
BMP-7
肾小球系膜细胞
活性氧
high glucose
BMP-7
glomerular mesangial cells
reactive oxygen species
