HINT2抑制脂多糖诱导的HaCat细胞炎症反应  

HINT2 Inhibits LPS-induced Inflammatory Response in HaCat Cells

作　　者：李翠[1] 秦锦博 孔舒欣 于洋[1] 梁栋 张斌[1] 尤伟 吕峰 Li Cui;Qing Jingbo;Kong Shuxin(Department of Breast Surgery,Henan University People's Hospital,Henan Provincial People's Hospital,Zhengzhou 450003,China)

机构地区：[1]河南大学人民医院,河南省人民医院乳腺外科,郑州450003

出　　处：《华中科技大学学报（医学版）》2020年第3期286-289,295,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：河南省卫生健康委员会科研资助项目(No.2018093)。

摘　　要：目的研究上调人三联组氨酸核苷酸结合蛋白2(histidine nucleotide binding 2,HINT2)表达对人上皮细胞HaCat细胞中脂多糖(lipopolysaccharides,LPS)诱导的炎症反应的影响。方法用慢病毒转染的方法构建HINT2过表达的HaCat细胞株HaCat^(HINT2);Western blot检测细胞HINT2的蛋白表达;LPS作用细胞6 h后,以Real-time PCR和ELISA分别检测培养细胞TNF-α、IL-6、IL-10的mRNA表达水平及其在细胞培养上清中的分泌量,Western blot检测PI3K/AKT通路及NF-κB通路的蛋白变化。结果Western blot证明高表达HINT2的HaCat细胞(HaCat^(HINT2))构建成功;LPS诱导后,HINT2高表达组的TNF-α、IL-6 mRNA表达低于对照组(HaCat^(control)),IL-10的mRNA表达高于对照组,差异均有统计学意义(均P<0.05);HINT2高表达组培养上清液中的TNF-α、IL-6分泌量低于对照组,IL-10分泌量高于对照组,差异均有统计学意义(均P<0.05);高表达HINT2可降低由LPS刺激引起的NF-κB信号通路p65蛋白磷酸化。结论高表达HINT2可以显著抑制HaCat细胞中LPS诱导的炎症反应以及NF-κB信号通路的激活。Objective To investigate the effect of up-regulation of histidine nucleotide binding 2(HINT2)expression on inflammatory response induced by lipopolysaccharides(LPS)in human epithelial HaCat cells.Methods Lenti-viruses were used to construct HaCat cell lines with over-expression of HINT2.Western blotting was used to detect the expression level of HINT2.RNA was extracted from HaCat cells after treatment with LPS for 6 h.Real-time PCR was used to detect the expression of TNF-α,IL-6 and IL-10 in cells.Meanwhile cell supernatant was collected.The expression of TNF-α,IL-6 and IL-10 in cell supernatant was detected by ELISA.The differences of the above indexes between the control and HINT2 group were compared.The activation of PI3 K/AKT pathway and NF-κB signaling pathway was detected by Western blotting.Results Western blotting confirmed that HINT2 highly expressed HaCat cells(HaCatHINT2)were successfully constructed.After LPS induction,TNF-αand IL-6 mRNA expression was lower in HaCatHINT2 than in control group(HaCatcontrol),and IL-10 expression in HaCatHINT2 was higher than that in control group,with significant difference(P<0.05).Meanwhile,the expression levels of TNF-αand IL-6 in the supernatant of HaCatHINT2 cells were lower than that of control group,and the expression of IL-10 was higher than that of control group,with significant difference(P<0.05).High-expression of HINT2 could inhibit the activation of NF-κB signaling pathway induced by LPS stimulation.Conclusion High-expression of Hint2 can significantly inhibit LPS-induced inflammation and activation of NF-κB signaling pathway in HaCat cells.

关 键 词：HINT2 炎症 HACAT 脂多糖 NF-ΚB信号通路 

分 类 号：R364.5[医药卫生—病理学]