白介素17对脂多糖诱导人支气管上皮细胞炎症损伤的影响  

Effect of IL-17A on the Inflammatory Injury Induced by LPS in Human Bronchial Epithelial Cell

作　　者：王梅芬 邹亚 郑培永[2] 景晓平[1] 何丽 郭盛[3] WANG Mei-fen;ZOU Ya;ZHENG Pei-yong;JING Xiao-ping;HE Li;GUO Sheng(Department of Traditional Chinese Medicine,Shanghai Children's Hospital,Shanghai Jiao Tong University,Shanghai,200040,China;Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai,200030,China;Department of Endocrinology,Genetics and Metabolism,Shanghai Children's Hospital,Shanghai Jiao Tong University,Shanghai,200040,China)

机构地区：[1]上海交通大学附属儿童医院中医科,上海200040 [2]上海中医药大学附属龙华医院,上海200030 [3]上海交通大学附属儿童医院内分泌遗传代谢科,上海200040

出　　处：《现代生物医学进展》2020年第10期1801-1805,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81774365);上海市科学技术委员会医学引导类科技支撑项目(16411964500);国家十三五传染病重大专项课题(2017ZX10305501-002)。

摘　　要：目的:探讨白介素17A(interleukin-17A,IL-17A)对脂多糖(lipopolysaccharide,LPS)诱导的人支气管上皮细胞(16-HBE)炎症损伤的影响及其可能机制。方法:体外培养16-HBE细胞系,予LPS、IL-17A进行干预,分为空白对照组、LPS组、IL-17A组、IL-17A+LPS组。采用酶联免疫吸附法(Enzyme linked immunosorbent assay,Elisa)测定细胞培养液上清中IL-4、IFN-γ、IL-6、IL-8等炎症因子的水平,蛋白印迹法(Western Blot,WB)检测细胞中丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPKs)信号通路相关蛋白:细胞外调节蛋白激酶(ERK)、P38蛋白激酶(P38)、c-Jun氨基末端激酶(JNK)的表达及其相应磷酸化蛋白(P-ERK、P-P38、P-JNK)的表达。体外培养16-HBE细胞系,予LPS、IL-17A以及ERK1/2抑制剂(U0126)、p38抑制剂(SB203580)和JNK抑制剂(SP600125),分为空白对照组、LPS+IL-17A组、IL-17A+LPS+UO126组、IL-17A+LPS+SB203580组、IL-17A+LPS+SP600125组。采用酶联免疫吸附法测定细胞培养液上清中IFN-γ、IL-4、IL-6、IL-8等炎症因子水平。结果:与空白对照组比较,LPS、IL-17A组,细胞上清中IL-6、IL-8的表达明显升高(P<0.01),IL-4的表达降低(空白组vs LPS组P<0.01,空白组vs IL-17A组P<0.05),细胞内磷酸化ERK、P38、JNK蛋白的表达明显增加(空白组vs LPS组P<0.05,空白组vs IL-17A组P<0.01)。IL-17A+LPS组细胞上清中IL-6、IL-8、IL-4水平及细胞内P-ERK、P-P38、P-JNK的表达较LPS、IL-17A组更高(P<0.05)。添加ERK、P38和JNK抑制剂后,与LPS+IL-17A组对比,IL-17A+LPS+U0126组、IL-17A+LPS+SB203580组和IL-17A+LPS+SP600125组细胞上清中IL-6、IL-8、IL-4水平下降(P<0.05)。结论:IL-17A可能通过上调IL-6、IL-8的表达加重LPS诱导的16-HBE细胞炎症损伤,MAPKs可能是这一过程中的重要信号转导通路。Objective: To investigate the effects of interleukin-17A(IL-17A) on lipopolysaccharide(LPS)-induced human bronchial epithelial cells(16-HBE) and their possible mechanisms. Methods: The 16-HBE cell line was cultured in vitro and intervened with LPS and IL-17A, and divided into blank control group, LPS group, IL-17A group and IL-17A+LPS group. The concentrations level of IL-4, IFN-γ, IL-6, IL-8 and other inflammatory factors in the supernatant of cell culture medium were determined by Enzyme linked immunosorbent assay(Elisa). The expression and activation of Mitogen-activated protein kinase(MAPKs) pathway related proteins: Phosphorylated extracellular regulated protein kinase(P-ERK), Extracellular regulated protein kinase(ERK), Phosphorylated P38 protein kinase(P-P38), P38 protein kinase(P38), Phosphorylated c-Jun N-terminal kinase(P-JNK), c-Jun N-terminal kinase(JNK) were detected by Western Blotting(WB). The 16-HBE cell line was cultured in vitro and intervened with LPS, IL-17A, and ERK inhibitor(U0126), p38 inhibitor(SB203580), and JNK inhibitor(SP600125), and divided into blank control group, LPS + IL-17A group, IL-17A + LPS + UO126 group, IL-17A + LPS + SB203580 group, IL-17A + LPS + SP600125 group. Enzyme-linked immunosorbent assay was used to determine the levels of inflammatory factors such as IFN-γ, IL-4, IL-6, and IL-8 in the cell culture supernatant. Results: Compared with the blank control group, the concentration level of IL-6 and IL-8 were significantly increased in the cellular supernatant(P<0.01), while the concentration level of IL-4 was decreased(con vs LPS P<0.01, con vs IL-17A P<0.05) in the supernatant with either IL-17A or LPS stimulation.The expression of P-ERK, P-P38 and P-JNK protein was significantly increased(con vs LPS P<0.05, con vs IL-17A P<0.01). the concentration level of IL-6 and IL-8 were significantly increased(P<0.01). The levels of IL-6, IL-8, IL-4 and the expression of P-ERK, P-P38,and P-JNK in the cell supernatant of IL-17A + LPS group were higher than those in LPS and IL-17

关 键 词：白介素17A 脂多糖 支气管上皮细胞 炎症因子 丝裂原活化蛋白激酶 

分 类 号：R-33[医药卫生] R322.34