| 王梅芬,邹 亚,郑培永,景晓平,何 丽,郭 盛.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[J].现代生物医学进展英文版,2020,(10):1801-1805. |
| 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 |
| Received:November 23, 2019 Revised:December 18, 2019 |
| DOI：10.13241/j.cnki.pmb.2020.10.001 |
| 中文关键词: 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 |
| 英文关键词: IL-17A LPS 16-HBE Inflammatory factors MAPKs |
| 基金项目: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） |
| Author Name | Affiliation | E-mail | | WANG Mei-fen | Department of Traditional Chinese Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, 200040, China | wangmf94@163.com | | ZOU Ya | 1 Department of Traditional Chinese Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, 200040, China
2 Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200030, China | | | ZHENG Pei-yong | Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200030, China | | | JING Xiao-ping | Department of Traditional Chinese Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, 200040, China | | | HE Li | Department of Traditional Chinese Medicine, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, 200040, China | | | GUO Sheng | Department of Endocrinology, Genetics and Metabolism, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, 200040, China | |
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| 中文摘要: |
| 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-?酌、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-?酌、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。 |
| 英文摘要: |
| ABSTRACT 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-17A group (P<0.05). After adding ERK, P38, and JNK inhibitors, compared with the LPS + IL-17A group, IL-17A + LPS + U0126 group, IL-17A + LPS + SB203580 group, and IL-17A + LPS + SP600125 group, the expression of IL-6, IL-8, IL-4 in cell supernatants has decreased (P<0.05). Conclusion: IL-17A might aggravate LPS-induced 16-HBE inflammatory injury by up-regulating IL-6 and IL-8 concentration level, and MAPKs may be an important signal transduction pathway in the process. |
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