环氧化酶-2抑制剂对人腹膜间皮细胞转化生长因子-β_1表达的影响  

Effects of cyclooxygenase-2 inhibitor on TGF-β_1 expression in human peritoneal mesothelial cells

作　　者：刘虹[1] 彭佑铭[1] 刘伏友[1] 

机构地区：[1]中南大学湘雅二医院肾内科,长沙410011

出　　处：《肾脏病与透析肾移植杂志》2003年第6期512-515,共4页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的:研究环氧化酶-2(COX-2)抑制剂对人腹膜间皮细胞(HPMC)转化生长因子-1(TGF-1)表达的影响。 方法:采用胰蛋白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。用COX-2抑制剂干预高糖(4.25%D-葡萄糖)和细菌脂多糖(LPS)刺激下的HPMC。采用逆转录多聚酶链式反应(RT-PCR)半定量分析HPMC中TGF-1 mRNA的表达。采用双抗夹心法酶联免疫吸附实验检测HPMC培养液中TGF-1蛋白质水平。 结果:HPMC在高糖和LPS的刺激下可明显上调TGF-1的表达(P<0.01),COX-2抑制剂干预组(20nmol/L,40nmol/L, 60nmol/L)明显下调TGF-1的表达(P<0.05)。 结论:COX-2抑制剂能够明显抑制在高糖和LPS的刺激下的HPMC TGF-1的基因表达,为临床用COX-2抑制剂防治腹膜透析患者的腹膜纤维化提供实验和理论依据。Objective: To investigate the effects of cyclooxygenase-2 (COX-2) inhibitor on TGF-b1 expression in human peritoneal mesothelial cells (HPMC). Methodology: HPMC were cultured from human omentum by an enzyme digestion method. The third passage HPMC stimulated by 4.25% D-glucose and lipopolysaccharide (LPS, 10mg/ml) were treated with 5nmol/L, 20nmol/L, 40nmol/L and 60nmol/L COX-2 inhibitor. The mRNA expression of TGF-b1 was determined by semi-quantification reverse transcriptive PCR (RT-PCR). The protein level of TGF-b1 in cultural supernatant was detected by a sandwich enzyme-linked immunosorbent assay. Results: The low level of TGF-b1 mRNA and protein were detected in confluent HPMC. With the treatment of 4.25% D-glucose and LPS, the level of TGF-b1 mRNA and protein were significantly upregulated (P<0.01). In treating cells, the introduction of COX-2 inhibitor (20nmol/L, 40nmol/L and 60 nmol/L) resulted in significant reduction of TGF-b1 mRNA and protein (P<0.05). Conclusion: COX-2 inhibitor inhibit the expression of TGF-b1 in HPMC stimulated by 4.25% D-glucose and LPS. These results may afford the experimental and academic evidence for the possible efficacy of COX-2 inhibitor for preventing peritoneal fibrosis in patients with peritoneal dialysis.

关 键 词：环氧化酶—2抑制剂 人腹膜间皮细胞 转化生长因子-β1 腹膜纤维化 胰蛋白酶消化法 RT-PCR 

分 类 号：R965[医药卫生—药理学]