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吴笛,蔺晓源,雷诗卉,姜帆,谭骏岚.基于TLR4/MyD88/NF-κB信号通路探讨紫菀散对慢性支气管炎气道炎症的干预作用[J].湖南中医药大学学报英文版,2023,43(7):1180-1187.[Click to copy
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| 基于TLR4/MyD88/NF-κB信号通路探讨紫菀散对慢性支气管炎气道炎症的干预作用 |
| 吴笛,蔺晓源,雷诗卉,姜帆,谭骏岚 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;中医内科重大疾病防治研究及转化教育部重点实验室, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察紫菀散对慢性支气管炎气道炎症的干预作用，并基于Toll样受体4（Toll-like receptors 4，TLR4）/髓分化因子88（myeloid differentiation factor 88，MyD88）/核转录因子-κB（nuclear factor kappa-B，NF-κB）信号通路探讨其作用机制。方法 60只健康成年雄性SD大鼠随机分为6组，每组10只，除正常组外均采用气管内滴注脂多糖（lipopolysaccharides，LPS）的方法建立慢性支气管炎模型。醋酸地塞米松组给予0.075 mg·kg-1醋酸地塞米松灌胃，每天1次；紫菀散低、中、高剂量组分别给予1.5、3.0、6.0 g·kg-1紫菀散灌胃，每天1次；模型组、正常组给予等量生理盐水灌胃。采用ELISA法检测支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中转化生长因子-β1（transforming growth factor-β1，TGF-β1）、白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-6（interleukin-6，IL-6）水平，采用细胞计数板法计数BALF中白细胞数量，采用HE染色、AB-PAS染色观察肺组织病理改变，采用免疫组化法检测肺组织TLR4、MyD88、NF-κB蛋白表达。结果 与正常组比较，模型组TGF-β1、IL-1β、IL-6水平均升高（P<0.01），白细胞总数和各种白细胞数量均增加（P<0.01），肺组织呈病理改变且气道杯状细胞增生评分升高（P<0.01），TLR4、MyD88、NF-κB蛋白表达均升高（P<0.01）；与模型组比较，醋酸地塞米松组及紫菀散各剂量组TGF-β1、IL-1β、IL-6水平均降低（P<0.05或P<0.01），白细胞总数和各种白细胞数量均降低（P<0.01），肺组织病理改变均减轻，气道杯状细胞增生评分均降低（P<0.05或P<0.01），TLR4、MyD88、NF-κB蛋白表达均减少（P<0.05或P<0.01）；与醋酸地塞米松组比较，紫菀散中、高剂量组TGF-β1水平均降低（P<0.05或P<0.01），紫菀散低剂量组白细胞总数和各种白细胞数量均增加（P<0.01），紫菀散中剂量组中性粒细胞数量、嗜酸性粒细胞数量均增加（P<0.01），紫菀散高剂量组淋巴细胞数量减少（P<0.01），紫菀散低、中剂量组肺组织病理改变均加重，紫菀散低剂量组气道杯状细胞增生评分升高（P<0.05），紫菀散低剂量组TLR4、MyD88、NF-κB蛋白表达均升高（P<0.05）。结论 紫菀散可能通过抑制TLR4/MyD88/NF-κB信号通路发挥抗慢性支气管炎气道炎症的作用。 |
| 关键词: 紫菀散 慢性支气管炎 气道炎症 TLR4/MyD88/NF-κB信号通路 转化生长因子-β1 白细胞介素-1β 白细胞介素-6 |
| DOI：10.3969/j.issn.1674-070X.2023.07.005 |
| Received:March 23, 2023 |
| 基金项目:湖南省自然科学基金科药联合基金项目（2021JJ80067）；湖南省“国内一流培育学科”药学开放基金项目（2021YX03）。 |
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| Intervention effects of Ziwan Powder on airway inflammation in chronic bronchitis based on TLR4/MyD88/NF-κB signaling pathway |
| WU Di,LIN Xiaoyuan,LEI Shihui,JIANG Fan,TAN Junlan |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Key Laboratory of Research on Prevention and Treatment of Major Diseases in Chinese Internal Medicine and Transformation of Achievements of Ministry of Education, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the intervention effects of Ziwan Powder on airway inflammation in chronic bronchitis, and to explore its mechanism based on toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. Methods Sixty healthy adult male SD rats were randomly divided into 6 groups, with 10 in each group. They were induced into chronic bronchitis models by instilling lipopolysaccharides (LPS) into the trachea, except for those of normal group. Dexamethasone Acetate group was given once daily gavage of 0.075 mg·kg-1 Dexamethasone Acetate. Once daily gavage of 1.5, 3.0 and 6.0 g·kg-1 extracts of Ziwan Powder were given to low-, medium-, and high-dose Ziwan Powder groups, respectively. The equal gavage of normal saline was administered to model group and normal group. The levels of transforming growth factor-β1 (TGF-β1), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. White blood cell count in BALF was checked by hemacytometer. Lung tissue pathological changes were observed by HE and AB-PAS staining. The protein expressions of TLR4, MyD88, and NF-κB in lung tissues were observed by immunohistochemistry staining. Results Compared with normal group, model group showed higher levels of TGF-β1, IL-1β, and IL-6 (P<0.01), elevated count of total white blood cells, macrophages, lymphocytes, neutrophil granulocytes, and eosinophil granulocytes (P<0.01), increased protein expression levels of TLR4, MyD88, and NF-κB (P<0.01), a higher pathological score of airway goblet cell hyperplasia (P<0.01), and obvious pathological changes in lung tissues. Compared with model group, Dexamethasone Acetate group and all Ziwan Powder groups shared lower levels of TGF-β1, IL-1β, and IL-6 (P<0.05 or P<0.01), reduced count of total white blood cells, macrophages, lymphocytes, neutrophil granulocytes and eosinophil granulocytes (P<0.01), decreased protein expression levels of TLR4, MyD88, and NF-κB (P<0.05 or P<0.01), a lower pathological score of airway goblet cell hyperplasia (P<0.05 or P<0.01), and improved pathological changes in lung tissues. Compared with Dexamethasone Acetate group, medium- and high-dose Ziwan Powder groups shared a lower level of TGF-β1 (P<0.05 or P<0.01); low-dose Ziwan Powder group showed elevated count of total white blood cells, macrophages, lymphocytes, neutrophil granulocytes, and eosinophil granulocytes (P<0.01), increased protein expression levels of TLR4, MyD88, and NF-κB (P<0.05), and an increased number of airway goblet cells (P<0.05); medium-dose Ziwan Powder group showed elevated count of neutrophil granulocytes and eosinophil granulocytes (P<0.01); high-dose Ziwan Powder group showed reduced count of lymphocytes (P<0.01); low-dose and medium-dose Ziwan Powder groups shared aggravated pathological changes in lung tissues. Conclusion Ziwan Powder may alleviate airway inflammation in chronic bronchitis by inhibiting the TLR4/MyD88/NF-κB signaling pathway. |
| Key words: Ziwan Powder chronic bronchitis airway inflammation TLR4/MyD88/NF-κB signaling pathway transforming growth factor-β1 interleukin-1β interleukin-6 |
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