乳酸盐腹膜透析液对人腹膜间皮细胞凋亡及bcl-2、bax表达和caspase-3活性的影响  

Effects of lactate peritoneal dialysis solution on apoptosis of human peritoneal mesothelial cells,expressions of bcl-2,bax and activity of caspase-3

作　　者：崔明姬[1] 王芳[2] 顾春梅[1] 黄金杰[1] 南光贤[3] 

机构地区：[1]吉林大学第一医院肾病科,吉林长春130021 [2]吉林大学药学院生物工程教研室,吉林长春130021 [3]吉林大学中日联谊医院脑血管病介入科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2009年第3期470-473,共4页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(200505197)

摘　　要：目的:探讨不同浓度乳酸盐腹膜透析液(L-PDS)对腹膜间皮细胞(HPMC)凋亡和bcl-2、bax表达及caspase-3活性的影响。方法:采用酶消化法从人腹膜组织中分离间皮细胞,建立稳定的体外培养模型,分别用浓度为1.50%、2.50%和4.25%的L-PDS与HPMC共同培养,同时设对照组。采用流式细胞术检测HPMC凋亡,采用RT-PCR法测定bcl-2及bax mRNA的表达,采用免疫荧光法检测caspase-3的活性。结果:与对照组比较,L-PDS各组HPMC凋亡率增加,bcl-2 mRNA的表达降低,bax mRNA表达升高,caspase-3活性增加,上述指标变化中2.50%及4.25%L-PDS组与对照组比较差异有显著性(P<0.05),4.25%L-PDS组与2.50%L-PDS组比较差异也有显著性(P<0.05),而1.50%L-PDS组与对照组比较差异无显著性(P>0.05)。结论:L-PDS可诱导HPMC凋亡,其机制可能是通过对bcl-2及bax mRNA表达的改变以及激活caspase-3活性而实现。Objective To study the effects of lactate peritoneal dialysis solution （L-PDS） with different concentrations on apoptosis of human peritoneal mesothelial cells （HPMC）, the expressions of bcl-2, bax and activity of caspase-3. Methods HPMC were separated using enzyme digestion and cultivated stably in vitro. After HPMC were co-cultivated with different concentrations （1.50%, 2.50%, 4.25%） L-PDS, flow cytometry was used to test the apoptosis of HPMC, RT-PCR was used to observe the expressions of bcl-2 and bax, fluorometric method was used to detect the activity of caspase-3. Results Compared with control group, L-PDS could induce the apoptosls of HPMC, especially in high concentration. With the increasing of L-PDS concentration, the expression of bcl-2 mRNA decreased, the expression of bax mRNA increased, the activity of caspase-3 raised. There were significant differences of the indexes mentioned above between 4.25%, 2.50% L-PDS groups and control group （P〈0.05）, there also was significant difference between 4.25% L-PDS group and 2.50% L-PDS group （P〈0.05）, but there was no significant difference between 1.50.% L-PDS group and control group （P〉0.05） . Conclusion L-PDS could induce HPMC apoptosis, which may be executed by alternating of the expressions of bcl-2, bax and activating of caspase-3.

关 键 词：乳酸盐腹膜透析液 腹膜间皮细胞 细胞凋亡 基因 BCL-2 bax caspase-3 

分 类 号：R459.5[医药卫生—治疗学]