高糖对人腹膜间皮细胞的增殖和损伤及分泌细胞因子的影响  

Effects of high glucose on the cell proliferation,damage and cytokine in human peritoneal mesothelial cells

作　　者：刘映红[1] 刘伏友[1] 张浩[2] 彭佑铭[1] 袁芳[1] 刘虹[1] 成梅初[1] 卓莉[1] 

机构地区：[1]中南大学湘雅二医院肾内科,长沙410011 [2]湘雅三医院肾内科,长沙410013

出　　处：《中南大学学报（医学版）》2006年第4期575-579,共5页Journal of Central South University ：Medical Science

摘　　要：目的:探讨高糖介导腹膜纤维化的可能机制。方法:原代培养的第3代人腹膜间皮细胞(HPMCs)分为不同时间(24,48h)的对照组(F12)和高糖组(F12+4%葡萄糖)。用MTT法测定细胞增殖,乳酸脱氢酶(LDH)法观察细胞损伤程度,采用酶联免疫法(ELISA)测定纤维连接蛋白(FN)、转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)蛋白水平以及采用RT-PCR测定FN,TGF-β1和纤溶酶原激活物抑制剂-1(PAI-1)mRNA表达。结果:①高糖明显抑制细胞增殖,24h和48h高糖组与同一时间点对照组比较,MTT吸光度值均明显下降(P<0.001或P<0.01);②高糖明显导致细胞损伤,24h和48h高糖组与同一时间点对照组比较,培养液中LDH含量均明显增加(均P<0.001);③高糖培养24,48h均使FN,CTGF和TGF-β1蛋白表达增加(P<0.05或P<0.001);④高糖使FN,TGF-β1和PAI-1mRNA表达均上调。结论:高糖能够抑制HPMCs增殖,损伤HPMCs,并刺激HPMCs分泌更多的TGF-β,CTGF,FN和PAI-1,从而使HPMCs细胞外基质生成增多、降解减少,最终导致腹膜纤维化的形成。Objective To determine the mechanism of peritoneal fibrosis of peritoneal mesothelial cells by high glucose. Methods The third passage human peritoneal mestothelial cells （ HPMCs ） protein expression of fibronectin （ FN ）, transforming growth factor-β1（ TGF-β1 ） and connective tissue growth factor （CTGF） were detected by ELISA. The mRNA expression of FN, TGF-β1 and PAl- 1 were detected by RT-PCR. Results High glucose suppressed the cell proliferation. The result of MTY showed that compared with the control group, the value of OD of high glucose groups at 24 or 48 h decreased significantly （ P 〈 0. 001 or 0.01 ） ; The cell damage was enhanced in high glucose groups, at 24 or 48 h compared with the control group at the same time （all P 〈0. 001 ）. The protein expressions of TGF-β1, CTGF and FN in supernate fluid of cell culture were significantly enhanced when high glucose stimulated the HPMCs in the high glucose groups at 24 or 48 h cornpared with the control group at same time （ P 〈 0.05 or 0.001 ）. The expressions of FN, TGF-β1 and PAl-1 mRNA were upregulated in 24 h high glucose group compared with that of 24 h control group. Conclusion High glucose crease of TGF-β1 , CTGF, FN and can suppresse the HPMC proliferation and damage HPMCs. InPAI-1 of HPMCs stimulated by high glucose can promote the svn-thesis and decreased degradation of extracellular matrix, which might be related with the mechanism of peritoneal fibrosis of peritoneal mesothelial cells by high glucose.

关 键 词：高糖 腹膜间皮细胞 增殖 损伤 细胞因子 

分 类 号：R692.5[医药卫生—泌尿科学]