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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 effect of cyclooxygenase-2 (COX-2) inhibitor on the expression of transforming growth factor-1 (TGF-1) in human peritoneal mesothelial cells (HPMC). Methods: Mesangial cells were isolated from human peritoneum by trypsin digestion, and a stable in vitro culture model was established. COX-2 inhibitors were used to intervene HPMC stimulated with high glucose (4.25% D-glucose) and bacterial lipopolysaccharide (LPS). Reverse transcription polymerase chain reaction (RT-PCR) semi-quantitative analysis of TGF-1 mRNA expression in HPMC. Double-antibody sandwich enzyme-linked immunosorbent assay TGF-1 protein levels in HPMC medium. Results: HPMC significantly upregulated the expression of TGF-1 (P <0.01) under the stimulation of high glucose and LPS, while down-regulated the expression of TGF-1 by 20 nmol / L, 40nmol / L and 60nmol / L of COX-2 inhibitor (P <0.05). CONCLUSION: The COX-2 inhibitor significantly inhibits the gene expression of HPMC TGF-1 stimulated by high glucose and LPS, providing experimental and theoretical evidence for the clinical use of COX-2 inhibitors in prevention and treatment of peritoneal fibrosis in patients undergoing peritoneal dialysis.