脂多糖对AR42J细胞Toll样受体7、9表达的影响  

Effect of lipopolysaccharide on toll like receptor 7 and toll like receptor 9 in AR42J cell lines

作　　者：张露艺[1] 唐国都[1] 唐曦平[1] 梁志海[1] 詹媛[1] 

机构地区：[1]广西医科大学第一附属医院,广西南宁530021

出　　处：《中华胰腺病杂志》2014年第3期163-166,共4页Chinese Journal of Pancreatology

基　　金：国家自然科学基金资助项目（81060043）;广西研究生教育创新计划资助项目（YCSZ2013034）

摘　　要：目的探讨Toll样受体（TLR）7、9在急性胰腺炎（AP）发病机制中的作用。方法用含1、10、100mg／L脂多糖的培养基处理正常大鼠胰腺腺泡细胞株AR42J，构建AP体外细胞模型，以不加脂多糖处理的细胞作为对照组。培养24h后收集细胞及培养上清液。采用RT-PCR法和蛋白质印迹法分别检测细胞TLR7、TLR9mRNA及蛋白的表达，ELISA法检测培养上清液中TNF-α、IL-10含量。结果对照组及1、10、100mg／L脂多糖处理组细胞TLR7mRNA表达量分别为0．12±0．09、0．28±0．06、0．49±0．04、0．78±0．04；TLR9mRNA表达量为0．06±0．02、0．32±0．03、0．56±0．14、0．84±0．12；TLR7蛋白表达量为0．04±0．01、0．26±0．05、0．49±0．04、0．77±0．16；TLR9蛋白表达量为0．10±0．14、0．62±0．23、1．21±0．26、1．75±0．13。培养上清液中TNF-α含量分别为（8．01±5．32）、（25．64±8．71）、（49．06±10．23）、（75．83±6．65）ng／L；IL-10含量分别为（155．54±25．47）、（105．16±10．49）、（69．36±8．19）、（14．07±9．06）ng／L。细胞TLR7和TLR9的mRNA及蛋白表达量、培养上清液中TNF-α含量均随脂多糖浓度升高而逐渐升高，而培养上清液中IL-10含量随脂多糖浓度增加逐渐下降，各组间的差异均具有统计学意义（P值均〈0．01）。结论脂多糖处理AR42J细胞后细胞的TLR7、TLR9基因表达明显上调，提示两者在AP疾病发展中可能发挥重要作用。Objective To investigate the roles of toll like receptor7 （TLR7） and toll like receptor 9 （TLR9） in the pathogenesis of acute panereatitis. Methods AR42J cells were treated by lipopolysaceharide at different dosages （0, 1, 10, 100 mg/L） , and cell model of acute panereatitis in vitro was established. AR42J cells without lipopolysaccharide treatment were as control. Cells and culture supematant were collected after 24 hours cultivation. TLR7, TLR9 mRNA and protein expressions were detected by RT-PCR and Western Blot, and levels of TNF-α, IL-10 in culture supernatant were measured by ELISA. Results The TLR 7 mRNA expression levels in control group, 1, 10, 100 mg/L lipopolysaeeharide group were 0.12 ± 0.09, 0.28 ± 0. 06, 0. 49 ± 0.04, 0.78 ± 0.04, and the TLR9 mRNA expression levels were 0. 06 ± 0. 02, 0.32 ± 0.03,0. 56 ± 0.14, 0.84 ± 0. 12 ; the TLR7 protein expression levels were 0. 04 ± 0.01, 0. 26 ± 0.05, 0.49 ± 0.04, 0.77 ± 0.16, and the TLR9 protein expression levels were 0.10 ± 0.14, 0.62 ± 0.23, 1.21 ± 0.26, 1.75 ± 0.13 ; the TNF-α levels in culture supematant were （ 8.01 ± 5.32 ）, （ 25.64 ± 8.71 ）, （49.06 ± 10.23 ）, （75.83 ± 6.65 ） ng/L, and the IL-10 levels were （ 155.54 ± 25.47 ） , （ 105.16 ± 10.49）, （69.36 ± 8.19）, （14.07 ± 9.06 ）ng/L. The expression levels of TLR7 and TLR9＇s mRNA, protein in cell, as well as the levels of TNF-α in cuhure supematant increased with the lipopolysaceharide concentration, whilethe levels of IL-10 in culture supernatant decreased with the lipopolysaccharide concentration, and the difference among these groups was statistically significant （ P 〈 0.01 ）. Conclusions The expressions of TLR7 and TLR9 in AR42J cells treated by using lipolysaccharide are obviously up-regulated, and it suggests that TLR7 and TLR9 may be vital in the pathogenesis of acute pancreatitis.

关 键 词：胰腺炎 TOLL样受体 脂多糖 损伤相关性分子模式 AR42J细胞 

分 类 号：R576[医药卫生—消化系统]