摘要
目的:基于沉默信息调节因子1(silencing information regulator 1,SIRT1)/腺苷酸活化蛋白激酶(adenosine activated monophosphate protein kinase,AMPK)信号通路探究壮宣饮对H1N1肺炎小鼠肺泡上皮细胞自噬和凋亡的影响。方法:随机选择6只小鼠作为对照组,其余小鼠通过鼻内滴入100μL甲型流感病毒(influenza A virus,IAV)[A/PuertoRico/8/34(PR8,H1N1)病毒]溶液建立H1N1肺炎小鼠模型。将造模成功小鼠随机平分为H1N1组、低-壮宣饮组(7.16 g·kg^(-1))、中-壮宣饮组(14.33 g·kg^(-1))、高-壮宣饮组(28.66 g·kg^(-1))、高-壮宣饮+Compound C组(28.66 g·kg^(-1)壮宣饮+0.25 mg·kg^(-1) SIRT1/AMPK信号通路抑制剂Compound C),每组均6只,每天上、下午各1次,连续5天,H1N1组和对照组给予等量生理盐水。ELISA法检测BALF中炎性因子水平;HE染色检测肺组织病理变化;分离和培养小鼠肺泡上皮细胞;流式细胞术检测肺泡上皮细胞凋亡;Western blot检测肺泡上皮细胞自噬蛋白、SIRT1/AMPK信号通路相关蛋白表达。结果:对照组肺组织结构正常,而H1N1组肺组织炎性细胞浸润到间质中、肺泡严重出血、塌陷、间质水肿、肺泡壁增厚,H1N1组较对照组γ-干扰素(interferon-γ,IFN-γ)、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平、肺泡上皮细胞凋亡率显著升高(P<0.05),AMPK、SIRT1蛋白水平、微管相关蛋白1轻链3(microtubule associated protein 1 light chain 3,LC3)-Ⅱ/Ⅰ、苄氯素1(benzyl chloride 1,Beclin 1)蛋白水平显著下降(P<0.05);与H1N1组相比,低-壮宣饮组、中-壮宣饮组、高-壮宣饮组IFN-γ、IL-6、TNF-α水平、肺泡上皮细胞凋亡率显著下降(P<0.05),AMPK、SIRT1蛋白水平、LC3-Ⅱ/Ⅰ、Beclin 1蛋白水平显著升高(P<0.05),且呈剂量相关性。Compound C逆转了高-壮宣饮的有利影响。结论:壮宣饮可能通过调控SIRT1/AMPK信号通路对H1N1肺炎小鼠肺泡上皮细胞自噬和凋亡起到改善作用。
Objective:To investigate the effect of Zhuangxuan drink on autophagy and apoptosis of alveolar epithelial cells in H1N1 pneumonia mice based on the silencing information regulator 1(SIRT1)/adenosine activated monophosphate protein kinase(AMPK) signaling pathway.Methods:Six mice were randomly selected as the control group,while the remaining mice were given 100 μL of influenza A virus(IAV) [A/Puerto Rico/8/34(PR8,H1N1)] virus solution through nasal drip to establish an H1N1 pneumonia mice model.Successfully modeled mice were randomly separated into H1N1 group,low-Zhuangxuan drink group(7.16 g·kg^(-1)),medium-Zhuangxuan drink group(14.33 g·kg^(-1)),high-Zhuangxuan drink group(28.66 g·kg^(-1)),and high-Zhuangxuan drink+Compound C group(28.66 g·kg^(-1) Zhuangxuan drink+0.25 mg·kg^(-1) SIRT1/AMPK signaling pathway inhibitor Compound C),with 6 mice in each group,once in the morning and once in the afternoon,for 5 consecutive days.The H1N1 group and control group were given equal amounts of physiological saline.ELISA method was applied to detect the levels of inflammatory factors in BALF.HE staining was applied to detect pathological changes in lung tissue.The mouse alveolar epithelial cells were isolated and cultured.Flow cytometry was applied to detect apoptosis of alveolar epithelial cells.Western blot was applied to detect the expression of autophagy proteins and SIRT1/AMPK signaling pathway related proteins in alveolar epithelial cells.Results:The lung tissue structure in the control group was normal,while in the H1N1 group,inflammatory cells infiltrated into the interstitium,and severe alveolar bleeding,collapse,interstitial edema,and thickening of alveolar walls were observed in the lung tissue.Compared with the control group,the levels of interferon-γ(IFN-γ),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and apoptosis rate of alveolar epithelial cells in the H1N1 group were obviously increased(P<0.05),and the protein levels of AMPK,SIRT1,microtubule associated protein 1 light chain 3(LC3)-Ⅱ/Ⅰ,a
作者
邹敏
韦杏
蓝毓营
张云
ZOU Min;WEI Xing;LAN Yuying;ZHANG Yun(Department of Pediatrics,Guangxi International Zhuang Medicine Hospital&Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine,Guangxi Nanning 530200,China;School of Zhuang Medicine,Guangxi University of Chinese Medicine,Guangxi Nanning 530200,China)
出处
《现代肿瘤医学》
CAS
2024年第17期3190-3196,共7页
Journal of Modern Oncology
基金
国家中医药管理局高水平中医药重点学科建设项目(少数民族医学-壮医学)(编号:zyyzdxk-2023164)
广西壮族自治区一流学科建设项目(编号:桂教科研[2022]1号)
广西壮族自治区中医药管理局自筹经费科研课题(编号:GZZC2020076,20210036)
广西壮族自治区中医药管理局广西中医药传承试点项目(编号:GXZYYXS-202304)
广西教育厅广西高校中青年教师基础能力提升项目(编号:2023KY0301)
广西国际壮医医院院级科研课题(编号:2023GZYJKT007)
广西中医药大学“高层次人才队伍建设三年行动计划”项目—国医大师黄瑾明学术思想与临床诊疗传承发展研究中心(编号:桂中医大党[2022]27号)。
关键词
壮宣饮
SIRT1/AMPK信号通路
肺泡上皮细胞
自噬
凋亡
Zhuangxuan drink
SIRT1/AMPK signaling pathway
pulmonary alveolar epithelial cells
autophagy
apoptosis
