脂肪细胞因子resistin诱导胰腺腺泡细胞促炎因子表达的研究

Induction of proinflammatory cytokines by adipocytokine resistin in pancreatic acinus cell line

目的:探讨脂肪细胞因子resistin诱导胰腺腺泡细胞促炎因子表达的作用。方法:大鼠胰腺腺泡AR42J细胞中分别加入不同终浓度的resistin蛋白(1、10、100μg/L)作用24 h。碘比色法测定培养液上清淀粉酶含量,酶联免疫吸附分析法检测培养液上清TNF-α与IL-6含量,实时定量PCR方法检测细胞TNF-α与IL-6 mRNA表达,Western印迹法检测细胞核NF-κB P65蛋白。结果:细胞培养液上清淀粉酶含量随resistin浓度升高而逐步上升,与对照组比较有统计学意义(P<0.01)。resistin浓度100μg/L时,细胞培养液上清TNF-α水平显著高于对照组[(155.1±26.2)ng/L比(122.8±10.9)ng/L,P<0.05]。resistin浓度10μg/L与100μg/L时,细胞培养液上清IL-6水平与对照组比较差异均有统计学意义[10μg/L:(28.1±2.0)ng/L比(23.0±2.0)ng/L,P<0.05;100μg/L:(29.8±2.4)ng/L比(23.0±2.0)ng/L,P<0.01]。resistin浓度100μg/L时,TNF-α与IL-6 mRNA表达均较对照组显著升高[TNF-α:(9.4±1.4)比(1.0±0.2),P<0.01;IL-6:(8.4±1.0)比(1.1±0.4),P<0.01],且细胞核NF-κB P65蛋白水平显著高于对照组(P<0.01)。结论:resistin可通过NF-κB途径诱导胰腺腺泡细胞促炎因子TNF-α与IL-6表达增加。

Objective To explore the effect of adipocytokine resistin on the induction of proinflammatory cytokines in pancreatic acinus cell line.Methods Rat pancreatic acinus AR42J cell line was stimulated by resistin at different dosages（1,10,100 μg/L） for 24 hours.The changes of amylase in culture medium were measured with chromatometry.The concentration of TNF-α and IL-6 in culture medium were detected with enzyme-linked immunosorbent assays.The mRNA expression of TNF-α and IL-6 were measured with real-time polymerase chain reaction.The expression of NF-κB P65 protein in the nucleus of AR42J cell lines were determined by Western blot.Results The concentration of amylase,TNF-α and IL-6 in culture medium increased significantly under the stimulation of resistin.The effect was maximum at 100 μg/L after 24-hour treatment（amylase and IL-6,P0.01;TNF-α,P0.05）.The mRNA expression of TNF-α and IL-6 were up-regulated significantly by 100 μg/L resistin treatment for 24 hours（P0.01）.Moreover,the nucleus NF-κB P65 protein also increased significantly（P0.01） under the same condition.Conclusions Our results suggest that resistin can induce the expression of proinflammatory cytokines,TNF-α and IL-6,through NF-κB pathway in pancreatic acinus cell line.