TLR9在脂多糖诱导的急性胰腺炎体外细胞模型中的作用

Role of TLR9 in pathogenesis of lipopolysaccharide-induced acute pancreatitis in an in vitro model

目的:探讨Toll样受体9(Toll-like receptor 9,TLR9)在急性胰腺炎发病机制中的作用.方法:不同浓度脂多糖(lipopolysaccharide,L P S)(0、1、10、100 m g/L)刺激A R42J细胞后,RT-PCR法与Western blot法分别检测TLR9、核因子-κB(nuclear factor-κB,NF-κB)P65 mRNA和蛋白质表达的变化,并分析TLR9、P65的相关性;ELISA法检测培养上清液白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)含量.结果:与空白对照组相比,LPS刺激后TLR9、P65 mR N A与蛋白质表达升高,呈浓度依赖性,各组间有显著性差异(mRNA:F=21.594,F=24.449;蛋白质:F=23.193,F=24.891,均P<0.01),相关性分析:r=0.942,r=0.900,均P=0.000,TLR9、P65呈正相关.上清液IL-1β、IL-6含量随LPS浓度增加而上升,组间差异有统计学意义(F=45.459,F=62.493,均P<0.01).结论:LPS刺激AR42J细胞诱导的炎症效应中,TLR9表达上调,可能通过激活NF-κB,从而促进炎症因子合成与分泌,参与急性胰腺炎发病机制.

AIM： To explore the role of Toll-like receptor 9 （TLR9） in the pathogenesis of acute pancreatitis. METHODS： After AR42J cells were stimulated with lipopolysaccharide at different concentrations （0, 1, 10, or 100 mg/L）, the mRNA and protein expression of TLR9 and nuclear factor κB （NF-κB） p65 was determined by RT-PCR and Western blot, respectively. The contents of interleukin-1β （IL-1β） and IL-6 proteins in culture supernatants were determined by ELISA. RESULTS： Lipopolysaccharide up-regulated TLR9 and P65 mRNA and protein expression in a dose-dependent manner （mRNA： F = 21.594, 24.449; protein： F = 23.193, 24.891, all P 〈 0.01）. There was a positive correlation between the mRNA and protein expression of TLR9 and that of P65 （mRNA： r = 0.942, protein： r = 0.900, both P = 0.000）. Lipopolysaccharide also increased the contents of IL-1β and IL-6 protein in a dose-dependent manner （F = 45.459, 62.493, both P 〈 0.01）. CONCLUSION： TLR9 may play a role in the pathogenesis of acute pancreatitis by up-regulating the expression of NF-κB.