TGFβ_1反义RNA对腹膜间皮细胞分泌细胞外基质的影响  

Effect of TGFβ_1 antisense RNA expression vector on synthesis and secretion of extracellular matrix in cultured human peritoneal mesothelial cells

作　　者：袁芳[1] 刘伏友[1] 刘映红[1] 段绍斌[1] 彭佑铭[1] 黄亮群[2] 

机构地区：[1]中南大学湘雅二医院肾内科,长沙410011 [2]中南大学医学遗传学国家重点实验室,长沙410078

出　　处：《中南大学学报（医学版）》2004年第5期548-551,共4页Journal of Central South University ：Medical Science

基　　金：教育部高等学校重点实验室访问学者基金 (教技司 2 0 0 0 ;1 70 );湖南省自然基金项目 (0 1JJY2 0 71 );湖南省卫生厅科学基金 (2 0 0 1 Y 39)

摘　　要：目的 :研究转化生长因子 β1(TGFβ1)反义RNA对体外培养的人腹膜间皮细胞 (HPMC)细胞外基质合成与分泌的影响。方法 :将反义TGFβ1基因真核表达载体转染HPMC ,用ELISA法检测转染细胞上清液中纤维连接蛋白 (FN)和Ⅰ型纤溶酶原激活物抑制剂 (PAI 1)的蛋白质水平 ;用半定量逆转录多聚酶链反应 (RT PCR)法检测转染细胞内FN ,PAI 1和胶原Ⅰ (ColⅠ )的mRNA表达 ,并与未转染HPMC对照组和转染空载体组比较其变化。结果 :TGFβ1反义RNA转染HPMC后 ,与对照组相比 ,转染 2 4h后 ,FN ,ColⅠ ,PAI 1mRNA分别下调 17% ,2 6 % ,9.6 % ;转染 4 8h后FN ,PAI 1蛋白含量亦明显下降 (P <0 .0 5 )。结论 :人反义TGFβ1基因真核表达载体可抑制HPMC合成细胞外基质 。Objective To determine the effect of transforming growth factor beta1(TGFβ 1) antisense RNA on the synthesis and secretion of extracellular matrix in human peritoneal mesothelial cells(HPMC). Methods The recombinant human TGFβ 1 antisense eukaryotic expression vector was transfected into HPMC.Fibronectin(FN) and plasminogen activator inhibitor-1(PAI-1) were detected by ELISA method. The expression of FN,collagenⅠ(colⅠ)and PAI-1 mRNA were detected by RT-PCR method.Results The mRNA of FN,ColⅠ,and PAI-1 in HPMC were repressed by TGFβ 1 antisense RNA 24 hours after the transfection in comparison with the control group,which decreased 17%,26%,and 9.6%,respectively. The protein of FN and PAI-1 were inhibited 48 hours after the transfection (P<0.05). Conclusion Recombinant human TGFβ 1 antisense eukaryotic expression vector can inhibit the synthesis and secretion of extracellular matrix in HPMC which may be effective in gene therapy for peritoneal fibrosis.

关 键 词：TGFΒ1 HPMC PAI-1 转染 FN 反义RNA 腹膜间皮细胞 分泌细胞 基因 真核表达载体 

分 类 号：R363[医药卫生—病理学]