Defensin α1对大鼠系膜细胞增殖的影响  

Recombinant human defensin α1 induces proliferation of rat glomerular mesangial cells in vitro

作　　者：张益民[1] 叶任高[1] 李幼姬[1] 李晓艳[1] 王昌云[1] 余学清[1] 董秀清[1] 

机构地区：[1]中山大学附属第一医院肾内科,教育部肾脏病重点实验室

出　　处：《中国病理生理杂志》2007年第9期1796-1800,共5页Chinese Journal of Pathophysiology

基　　金：教育部博士学科点专项科研基金资助项目(No.2000056916)

摘　　要：目的:探讨defensin α1对体外培养的大鼠肾小球系膜细胞株1097增殖的影响以及可能的机制。方法:利用MTT掺入观察不同浓度的基因重组人类defensin α1在不同作用时间对系膜细胞增殖的影响,选择能够刺激系膜细胞生长的浓度及作用时间给予U0126预孵育,观察能否阻断defensin α1促系膜细胞增殖作用,使用Western blotting了解defensin α1对系膜细胞ERK1/2磷酸化及Ⅳ型胶原合成的影响。结果:Defensin α1在3-20mg/L范围内对系膜细胞具有促生长作用,12h达到峰值(P<0.01)。当浓度大于20mg/L时可抑制系膜细胞的生长,浓度在15-25μmol/L U0126可以阻断defensin α1对系膜细胞的促生长作用。浓度为3mg/L defensin α1刺激5min后可使ERK1/2磷酸化明显增加,作用12h后系膜细胞的IV胶原蛋白表达高于对照组并持续到48h(P<0.01)。结论:一定浓度的defensin α1可以促进系膜细胞的增殖和细胞外基质的合成,这种促增殖作用可能是通过MAPK途径实现的。AIM： To study the effects and mechanism of recombinant human defensin α1 on cell proliferation in cultured rat glomerular mesangial cells. METHODS： The influences of defensin α1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay. The different concentrations of U0126, signal - regulated protein kinase （MEK） inhibitor, were added into the culture mediums of mesangial cells to do blocking test. Incubated with a final concentration of 3 mg/L defensin α1, the phosphorylation of extracellular signal regulated kinase （ERK） 1/2 and type Ⅳ collagen of mesangial cells in different times were evaluated by Western blotting. RESULTS： Defensin a 1 at 3 - 20 mg/L enhanced proliferation of rat glomerular mesangial cells. The incubation times for the maximum effect on proliferation was 12 h （ P 〈 0. 01 ）, whereas defensin α1 concentration 〉 20 mg/L decreased cell proliferation. The cell proliferation induced by defensin α1 was inhibited by U0126. Stimulation of the cells with defensin α1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK. After 12 h incubation with defensin α1, an increase in type Ⅳ collagen was observed by Western blotting and continued to increase at 24 h and 48 h （P 〈 0.01 ）. CONCLUSION： Defensin α1 enhances rat glomeular mesangial cell proliferation and induces type Ⅳ collagen production by MAPK signaling pathway.

关 键 词：防御素类 肾小球系膜细胞 细胞增殖 

分 类 号：R363[医药卫生—病理学]