热休克转录因子2通过HMGB1-TLR4-NF-κB信号促进克罗恩病炎症反应的作用机制研究

doi:10.12092/j.issn.1009-2501.2019.04.002

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (4): 369-375.doi: 10.12092/j.issn.1009-2501.2019.04.002

热休克转录因子2通过HMGB1-TLR4-NF-κB信号促进克罗恩病炎症反应的作用机制研究

阮水良，官俏兵

嘉兴市第二医院，嘉兴 314000，浙江

收稿日期:2018-09-21 修回日期:2018-10-10 出版日期:2019-04-26 发布日期:2019-05-01
通讯作者: 官俏兵，通信作者，女，学士，主任医师，研究方向：炎症性肠病。 E-mail：qbguan@126.com
作者简介:阮水良，男，硕士，副教授、副主任医师，研究方向：胃肠病学。 E-mail：ruanguan@hotmail.com
基金资助:
浙江省科技厅实验动物项目（2017C37174）；浙江省医药卫生一般项目（2017KY653）

Mechanism of heat shock transcription factor 2 promotes inflammatory response in Crohn's disease through HMGB1-TLR4-NF-κB signaling

RUAN Shuiliang，GUAN Qiaobing

The Second Hospital of Jiaxing, Jiaxing 314000, Zhejiang, China

Received:2018-09-21 Revised:2018-10-10 Online:2019-04-26 Published:2019-05-01

摘要/Abstract

摘要：

目的:研究热休克转录因子2（HSF2）通过HMGB1-TLR4-NF-κB信号促进小鼠克罗恩病炎症反应的作用机制。方法:2,4,6-三硝基苯磺酸（TNBS）构建小鼠克罗恩病模型，将小鼠分为正常组，模型组，对照组，实验组。对照组采用高迁移率族蛋白1（HMGB1）抗体处理阻断HMGB1信号，对照组和实验组均采用重组HSF2干预。观察小鼠一般生活状况（包括：生存状态、体质量状态、大便性状、进食饮水状态、毛发光泽度、精神状态和活动状态），小鼠疾病活动指数（DAI）评估小鼠疾病情况，苏木精-伊红（HE）染色检测小鼠肠组织病理改变，免疫组织化学染色（IHC）检测小鼠肠组织中p-65的表达，酶联免疫吸附法（ELISA）检测小鼠结肠组织中炎症因子白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）的水平，蛋白免疫印迹法（Western blot）检测结肠组织中HMGB1、Toll样受体4（TLR4）、NF-κB（p-P65)、髓样分化因子（MyD88）的表达水平。结果:TNBS成功诱导小鼠克罗恩病，实验组小鼠一般生活状态较差，且DAI评分显著高于对照组、模型组，具有统计学意义（P<0.05）。HE染色显示实验组小鼠结肠黏膜糜烂、水肿、炎症反应程度高于模型组和对照组。实验组小鼠结肠组织中炎症因子IL-1β、TNF-α、IL-6的水平显著高于模型组和对照组，差异具有统计学意义（P<0.05）。实验组组织中HMGB1、TLR4、NF-κB（p-P65)、MyD88的表达水平显著高于模型组和对照组，差异具有统计学意义（P<0.05）。结论:HSF2可以通过激活HMGB1-TLR4-NF-κB信号促进小鼠克罗恩病炎症反应。

关键词: 热休克转录因子2, 克罗恩病, 炎症反应, 高迁移率族蛋白1, Toll样受体4, NF-κB

Abstract:

AIM: To study the mechanism of heat shock transcription factor 2 (HSF2) promoting the inflammatory response of Crohn's disease in mice through HMGB1-TLR4-NF-κB signaling. METHODS: The model of Crohn's disease mice was established by 2,4,6-three nitrobenzene sulfonic acid (TNBS). Mice were divided into normal group, model group, control group and experimental group. The control group used high mobility group protein 1 (HMGB1) antibody to block the HMGB1 signal. The control group and the experimental group were treated with recombinant HSF2. To observe the general living conditions of mice (including survival, body weight, stool traits, eating and drinking state, hair glossiness, mental state and activity state), the disease activity index (DAI) of mice was used to evaluate the condition of the mice. Hematoxylin eosin (HE) staining was used to detect the pathological changes of intestinal tissue in mice. Immunohistochemical staining (IHC) was used to detect the expression of P-65 in intestinal tissue of mice. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the colon tissue of mice. Western-Blot was used to detect the expression level of HMGB1, Toll like receptor 4 (TLR4), NF-κB (p-P65) and myeloid differentiation factor (MyD88) in colonic tissue. RESULTS:TNBS induced mouse Crohn's disease successfully. The general living condition of the experimental group was poor, and the DAI score was significantly higher than that of the control group and the model group, with statistical significance (P<0.05). HE staining showed that the degree of erosion, edema and inflammatory reaction of the colon mucosa in the experimental group was higher than that in the model group and the control group. The levels of inflammatory factors IL-1β, TNF-α, IL-6 in the colon tissue of the experimental group were significantly higher than those of the model group and the control group (P<0.05). The expression level of HMGB1, TLR4, NF-κB (p-P65) and MyD88 in the tissues was significantly higher than that in the model group and the control group (P<0.05). CONCLUSION:HSF2 can promote the inflammatory response of Crohn's disease in mice by activating HMGB1-TLR4- NF-κB signaling.

Key words: heat shock transcription factor 2, Crohn's disease, inflammatory response, high mobility group box protein 1, Toll like receptor 4, NF-κB

中图分类号:

R574

阮水良，官俏兵. 热休克转录因子2通过HMGB1-TLR4-NF-κB信号促进克罗恩病炎症反应的作用机制研究[J]. 中国临床药理学与治疗学, 2019, 24(4): 369-375.

RUAN Shuiliang，GUAN Qiaobing. Mechanism of heat shock transcription factor 2 promotes inflammatory response in Crohn's disease through HMGB1-TLR4-NF-κB signaling[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(4): 369-375.

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热休克转录因子2通过HMGB1-TLR4-NF-κB信号促进克罗恩病炎症反应的作用机制研究

Mechanism of heat shock transcription factor 2 promotes inflammatory response in Crohn's disease through HMGB1-TLR4-NF-κB signaling

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