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  

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

作　　者：王彬[1] 张爱青[1] 龚晶[1] 郑君[1] 曾智凤[1] 甘卫华[1] 

机构地区：[1]南京医科大学第二附属医院儿肾科,江苏南京210003

出　　处：《南京医科大学学报（自然科学版）》2010年第12期1696-1699,1744,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省"六大人才高峰"项目资助(06-B-020)

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、0.16μmol/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0.04μmol/L至0.16μmol/L的呈浓度依赖抑制PDGF-BB诱导的MC增殖(P<0.05)。0.08μmol/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)。0.08μmol/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。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.

关 键 词：蟾蜍灵 系膜细胞 P21 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]