色素上皮衍生因子对高糖诱导人肾小球系膜细胞转化生长因子-β1／Smad信号转导途径的影响  

Influence of pigment epithelium-derived factor on transforming growth factor-β1/Smad pathway in human glomerular mesanglal cells cultured in high glucose

作　　者：毛拓华[1] 黄汉奇[1] 洪练[1] 李竞[1] Mao Tuohua;Huang Hanqi;Hong Lian(Department of Endocrinology,Renmin Hospital of Wuhan University,Wuhan 430060,Chin)

机构地区：[1]武汉大学人民医院内分泌科,430060

出　　处：《临床内科杂志》2018年第7期488-492,共5页Journal of Clinical Internal Medicine

摘　　要：目的观察色素上皮衍生因子（PEDF）对高糖诱导人肾小球系膜细胞（HMCs）转化生长因子-β1（TGF-β1）／Smad信号转导途径的影响。方法体外培养HMCs并将其分为正常对照组（5．6mmol／L葡萄糖）、渗透压对照组（5．6mmol／L葡萄糖＋24．4mmol／L甘露醇）、高糖组（30mmol／L葡萄糖）、干预1组（30mmol／L葡萄糖＋10nmol／LPEDF）、干预2组（30mmol／L葡萄糖＋40nmol／LPEDF）和干预3组（30mmol／L葡萄糖＋100nmol／LPEDF），分别作用24h后收集样本，采用半定量逆转录聚合酶链反应（RT—PCR）检测TGF-β1、纤维连接蛋白（FN）及Smad7 mRNA表达，Western blot检测Smad7蛋白的表达，酶联免疫吸附试验（ELISA）检测TGF-β1、FN蛋白水平，免疫细胞化学检测p-Smad2／3蛋白的表达。结果与正常对照组比较，高糖组TGF-β1、FN表达明显增加，Smad7表达明显减少（P〈0．05或P〈0．01），p-Smad2／3蛋白表达增加，而渗透压对照组与正常对照组比较无明显改变。与高糖组比较，PEDF干预组TGF-β1、FN表达减少，Smad7表达增加（P〈0．05或P〈0．01），p-Smad2／3蛋白表达呈下降趋势。结论PEDF能抑制高糖诱导的TGF-β1、FN和p-Smad2／3蛋白过度表达及上调Smad7表达。PEDF可能通过影响TGF-β1／Smad信号转导途径达到抗纤维化的作用，延缓糖尿病肾病的进程。Objective To investigate the influence of pigment epithelium-derived factor（PEDF） on transforming growth factor -β1 （TGF-β1）/Smad pathway of human glomerular mesangial cells（HMCs） cultured in high glucose. Methods Cultured HMCs were divided into the following groups：normal control group（5.6 mmol/L glucose）, osmotic pressure control group（5.6 mmol/L glucose ＋ 24.4 mmol/L mannitol ）, high glucose group（ 30 mmol/L glucose） ,intervention group 1 （30 mmoL/L glucose ＋ 10 nmol/L PEDF） , intervention group 2 （ 30 mmol/L glucose ＋ 40 nmol/LPEDF） and intervention group3 （ 30 mmol/L glucose ＋ 100 nmol/L PEDF）. After 24 h,the expression of TGF-β1 ,ibronectin（FN） and Smad7 mRNA of HMCs were examined by semi-quantitative RT-PCR. The protein expression of Smad7 was examined by Western blotting while TGF-β1 and FN were examined by ELISA. The p-Smad2/3 protein was detected by immunocytochemistry. Results Compared to normal control group, the mRNA and protein expressions of TGF-β1 and FN were remarkably increased in the cells exposed to high glucose, while the Smad7 expression was down-regulated （P 〈 0.05 or P 〈0. 01 ）. A significant increase of p-Smad2/3 protein was also observed in the cells exposed to high glucose compared to normal control. There was no significant change in the osmotic pressure control group compared with the normal control group. The expressions of TGF-β1 and FN mRNA and protein were decreased in the cells exposed to high glucose in the presence of PEDF, while the Smad7 mRNA and protein expressions were up-regulated（ P 〈0. 05 or P 〈 0.01 ）. In parallel, a significant decrease of p-Smad2/3 protein was observed in the cells exposed to high glucose in the presence of different concentrations of PEDF. Conclusion PEDF inhibited high glucose induced TGF-β1, FN and p-Smad2/3 overexpression in HMCs, while upregulated Smad7. Therefore, PEDF may play a salutary role in diabetic kidney disease by its anti-fibrogenic activities.

关 键 词：色素上皮衍生因子 转化生长因子-Β1 纤维连接蛋白 P—Smad2/3 SMAD7 

分 类 号：R656.4[医药卫生—外科学]