蟾蜍灵调控系膜细胞增殖与细胞周期激酶抑制剂p21的关系
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江苏省“六大人才高峰”项目资助(06-B-020)


Role of cyclin kinase inhibitor p21 in the regulation of bufalin on mesangial cell proliferation
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    摘要:

    目的: 观察蟾蜍灵对大鼠系膜细胞(mesangial cell,MC) p21表达的影响,探讨其调控系膜细胞增殖的机制-方法:MTT法观察不同浓度蟾蜍灵(0.01-0.04-0.08-0.16 μmol/L)对PDGF-BB(20 ng/ml)诱导的MC 增殖的影响;流式细胞术测定MC细胞周期的变化;RT-PCR法测定p21基因表达的改变;Western blot法测定p21蛋白水平变化-结果:PDGF-BB刺激MC12 h后,加入不同浓度蟾蜍灵干预24 h,蟾蜍灵从0.04 μmol/L至0.16 μmol/L的呈浓度依赖抑制PDGF-BB诱导的MC增殖(P < 0.05)-0.08 μmol/L蟾蜍灵干预24 h后,可使PDGF-BB诱导的MC G0/G1期细胞比例升高,使S期细胞比例下降,且上调P21mRNA和蛋白水平的表达(P < 0.05)-0.08 μmol/L蟾蜍灵对正常系膜细胞无明显影响(P > 0.0.5)-结论:蟾蜍灵可能通过上调细胞周期激酶抑制剂p21的表达来抑制PDGF-BB诱导的MC的增殖-

    Abstract:

    Objective: To investigate the role of p21 in the regulation of bufalin on the mesangial cell (MC) proliferation. Methods: The effects of different concentrations of bufalin (0.01,0.04,0.08,0.16 μmol/L) on MC proliferation induced by PDGF-BB(20 ng/ml) was evaluated by MTT test; Flow cytometry and RT-PCR were utilized for cell cycle analysis and p21 DNA expression,,respectively. Western blot was used to analyze protein expression of p21. Results: After twelve hours of PDGF-BB stimulation, MC was cultured with different concentrations of bufalin from 0.04 μmol/L to 0.16 μmol/L for 24 hours, respectively. Proliferation of GMC induced by 20 ng/ml PDGF-BB was inhibited significantly by bufalin in a dose-dependent manner(P < 0.05). Similar results were observed in 0.08 μmol/L bufalin, which blocked the PDGF-BB-induced progression from G0/G1 to S phase of the cell cycle in synchronized cells(P < 0.05). In addition, we found that bufalin reversed PDGF-BB-induced down-regulation of cyclin-dependent kinase inhibitor p21 on both gene and protein level(P < 0.05). However, 0.08μmol/L bufalin had no effects on normal mesangial cell proliferation (P > 0.05). Conclusion: Bufalin may inhibit the PDGF-BB-induced MC proliferation by increase of p21 expression.

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王 彬,张爱青,龚 晶,郑 君,曾智凤,甘卫华.蟾蜍灵调控系膜细胞增殖与细胞周期激酶抑制剂p21的关系[J].南京医科大学学报(自然科学版),2010,(12):1696-1699

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  • 收稿日期:2010-05-28
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