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张鑫,王孟清,罗银河,胡燕,丁伊,江智豪.RSV诱导哮喘小鼠树突细胞自噬机制及五虎汤的干预作用[J].湖南中医药大学学报,2021,41(1):39-45[点击复制] |
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| RSV诱导哮喘小鼠树突细胞自噬机制及五虎汤的干预作用 |
| 张鑫,王孟清,罗银河,胡燕,丁伊,江智豪 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学中医学国内一流建设学科, 湖南 长沙 410208) |
| 摘要: |
| 目的 研究PI3K/Akt/mTOR信号通路对呼吸道合胞病毒（respiratory syncytial virus，RSV）诱导哮喘小鼠树突细胞（dendritic cells，DC）自噬的调控机制及五虎汤的干预作用。方法 用RSV滴鼻联合雾化鸡卵清蛋白（ovalbumin，OVA）激发的方法制备哮喘小鼠模型，造模后的小鼠分为模型组、五虎汤低剂量组、五虎汤中剂量组、五虎汤高剂量组、地塞米松组、雷帕霉素组、氯喹组，每组10只，并设空白组（10只）对照。实验结束后PAS、Masson染色观察评价肺组织病变情况，并计算气道黏液储备指数与胶原沉积指数；电镜观察自噬小体；Western blot检测DC自噬相关蛋白及对PI3K/Akt/mTOR信号通路中关键蛋白PI3K、p-Akt、p-mTOR表达的影响。结果 气道反应性结果显示造模成功。病理染色显示模型组小鼠气道黏液储备指数与胶原沉积指数较空白组显著增高，差异有统计学意义（P<0.01）；与模型组相比，五虎汤各剂量组及其他药物组哮喘小鼠气道黏液储备指数与胶原沉积指数降低，差异有统计学意义（P<0.01）。电镜下观察小鼠肺组织自噬小体显示模型组相较于空白组中自噬小体数量明显增加，各治疗组的自噬小体较模型组增加。Western blot显示，自噬蛋白LC3Ⅱ/LC3Ⅰ在五虎汤各剂量组以及雷帕霉素组中的表达显著增高，与模型组相比差异有统计学意义（P<0.01）；氯喹组LC3Ⅱ/LC3Ⅰ水平较模型组降低（P<0.01）；地塞米松组与模型组比较，LC3Ⅱ/LC3Ⅰ水平相对表达无统计学差异（P>0.05）；通路关键蛋白表达水平显示，五虎汤各剂量组及雷帕霉素组中PI3K、p-Akt和p-mTOR的蛋白表达水平与模型组相比明显降低，差异有统计学意义（P<0.01，P<0.05）。结论 RSV诱导哮喘小鼠树突细胞自噬水平上升，PI3K/Akt/mTOR信号通路被活化，五虎汤通过抑制PI3K/Akt/mTOR信号通路，进一步上调树突细胞的自噬，减轻气道重塑。 |
| 关键词: 儿童哮喘 五虎汤 呼吸道合胞病毒 树突细胞 细胞自噬 PI3K/Akt/mTOR |
| DOI：10.3969/j.issn.1674-070X.2021.01.008 |
| 投稿时间：2020-07-03 |
| 基金项目:国家自然科学基金面上项目（81774368）；湖南省自然科学基金科卫基金（2018JJ6114）；湖南省研究生科研创新资助项目（CX20190580）；湖南中医药大学中医学国内一流学科建设学科项目（4901-020002002）。 |
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| The Mechanism of RSV-induced Autophagy of Dendritic Cells in Asthmatic Mice and the Interventional Effect of Wu-hu Decoction |
| ZHANG Xin,WANG Mengqing,LUO Yinhe,HU Yan,DING Yi,JIANG Zhihao |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Domestic First-class Discipline Construction Project of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To study the regulation mechanism of PI3K/Akt/mTOR signaling pathway on RSV-induced autophagy of dendritic cells in asthmatic mice and the intervention effect of Wuhu Decoction. Methods The mouse model of asthma was established by RSV nasal drop combined with ovalbumin (OVA) atomization. After modeling, mice were divided into the model group, the Wu-hu Decoction low-dose group, the Wu-hu Decoction medium-dose group, the Wu-hu Decoction high-dose group, the dexamethasone group, the rapamycin group and the chloroquine group, with 10 mice in each group, and a blank group (10 mice). After the end of the experiment, PAS staining and Masson staining were used to observe and evaluate the pathological changes of lung tissue, and the airway mucus reserve index and collagen deposition index were calculated; autophagy was observed by electron microscopy; Western blot was used to detected the autophagy-related proteins of DC and their effects on the expression of PI3K, p-Akt, and p-mTOR in the PI3K/Akt/mTOR signaling pathway. Results The airway reactivity results showed that the model was successful; pathological staining showed that the airway mucus reserve index and collagen deposition index of mice in the model group were significantly higher than the blank group, and the difference was statistically significant (P<0.01); compared with the model group, the airway mucus reserve index and collagen deposition index of asthma mice in Wu-hu Decoction each dose groups and other drug groups decreased, with statistical significance (P<0.01). Observation of autophagosomes in mouse lung tissue under electron microscope showed that the number of autophagosomes in the model group was significantly increased compared with the blank group, and the autophagosomes in each treatment group increased compared with the model group. Western blot showed that the autophagy protein LC3Ⅱ/LC3Ⅰin Wu-hu Decoction each dose groups and the rapamycin group were significantly higher than the model group, with statistical significance (P<0.01); the LC3Ⅱ/LC3Ⅰlevel of the chloroquine group was lower than the model group (P<0.01); there was no significant difference in the relative expression of LC3Ⅱ/LC3Ⅰbetween the dexamethasone group and the model group (P>0.05); the key proteins expression levels of pathway showed that the protein expressions of PI3K, p-Akt and p-mTOR in Wu-hu Decoction dose groups and rapamycin group were significantly lower than the model group, and the difference was statistically significant (P<0.01, P<0.05). Conclusion The autophagy level of DC cells in RSV-induced asthma mice increased, and the PI3K/Akt/mTOR signaling pathway was activated. Wuhu Decoction inhibited the PI3K/Akt/mTOR signaling pathway to further up-regulate DC cell autophagy and reduce airway remodeling. |
| Key words: childhood asthma Wu-hu Decoction respiratory syncytial virus dendritic cells cell autophagy PI3K/Akt/mTOR |
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