CRF通过CRF2受体介导肠上皮细胞中TLR4的表达  

Activation of TLR4 by CRF in human intestinal epithelial cells is mediated by the CRF2 receptor

作　　者：杨丽[1] 郑鹏远[1] 刘志强[1] 

机构地区：[1]郑州大学第二附属医院消化内科郑州大学医学微生态学研究所,河南省郑州市450014

出　　处：《世界华人消化杂志》2012年第19期1763-1767,共5页World Chinese Journal of Digestology

基　　金：973计划前期研究专项课题基金资助项目;No.2011CB512006~~

摘　　要：目的:研究促肾上腺皮质释放因子(CRF)介导肠上皮细胞中Toll样受体4(toll-like receptor4,TLR4)的表达,并探讨其可能通过的受体途径.方法:常规培养人结肠上皮细胞株HT-29细胞,将HT-29细胞分为正常对照组(不加刺激剂),脂多糖(lipopolysaccharide,LPS)刺激组(LPS20g/L刺激24h),促肾上腺皮质释放因子(corticotrophin-releasing factor,CRF)刺激组(CRF20g/L刺激24h),CRF+LPS刺激组(预先CRF20g/L刺激12h,更换细胞液后再与LPS20g/L刺激12h),CRF+Antalarmin 组(CRF与Antalarmin20g/L共刺激24h),CRF+LPS+Antalarmin组(CRF与Antalarmin20g/L共刺激12h后再以LPS刺激12h),CRF+Astressin2B组(CRF与Astressin2B20g/L共刺激24h),CRF+LPS+Astressin2B组(CRF与Astressin2B20g/L共刺激12h后再以LPS刺激12h).刺激结束后,收取各组HT-29细胞,RT-PCR法和免疫印迹法检测各组上皮细胞中TLR4mRNA和蛋白的表达.ELISA法检测各组细胞上清液中IL-8的表达.结果:CRF可以诱导人结肠上皮细胞株HT-29细胞中TLR4表达导致IL-8分泌增多(P<0.05),C R F1受体拮抗剂不能有效地阻滞C R F对TLR4的诱导(P>0.05,CRF+LPS组vs CRF组),CRF2受体拮抗剂可阻滞CRF对TLR4的诱导(P<0.05,CRF+LPS组vs CRF组).结论:CRF通过CRF2受体通路介导肠上皮细胞中TLR4的表达.AIM： To investigate the effect of corticotrophin- releasing factor （CRF） on the expression of toll- like receptor 4 （TLR4） in human intestinal epi- thelial cell line HT-29. METHODS： HT-29 cells were divided into eight groups： non-treated group, LPS group （treated with 20 μg/L LPS for 24 h）, CRF group （treated with 20 μg/L CRF for 24 h）, LPS plus CRF group （pretreated with 20 μg/L CRF for 12 h and then treated with 20 μg/L LPS for 12 h）, Astressin 2B plus CRF group （pretreated with 20 μg/L As- tressin 2B for 12 h and then treated with 20 μg/L CRF）, Antalarmin plus CRF group （pretreated with 20 μg/L Antalarmin for 12 h and then treated with 20 μg/L CRF）, Astressin 2B plus LPS group （pretreated with 20 μg/L Astressin 2B for 12 h and then treated with 20 μg/L LPS）, and An- talarmin plus LPS group （pretreated with 20 μg/ L Antalarmin for 12 h and then treated with 20 μg/L LPS）. The expression of TLR4 mRNA and protein was detected by reverse transcription- polymerase chain reaction （RT-PCR） and Western blotting, respectively. The secretion of interleu- kin-8 in the culture supernatants was determined by enzyme-linked immunosorbent assay （ELISA）. RESULTS： CRF could induce the expression of TLR4 in HT-29 cells and result in increased interleukin-8 secretion （P 〈 0.05）. CRFR2 antago- nist Astressin 2B inhibited the expression of LR4 （P 〈 0.05, CRF＋LPS group vs CRF group）, while CRF1 antagonist Antalarmin had no signifi- cant effect on the expression of TLR4 （P 〉 0.05, CRF＋LPS group vs CRF group）. CONCLUSION： The induction of TLR4 expres- sion by CRF in human intestinal epithelial cells is mediated by the CRF2 receptor.

关 键 词：促肾上腺皮质释放因子 TOLL样受体 肠上皮细胞 

分 类 号：R363[医药卫生—病理学]