摘要
目的探索麻杏石甘汤通过影响肺组织糖酵解而治疗病毒性肺炎的作用机制。方法通过鼻腔接种法建立甲型H1N1流感病毒株A/PR/8/34感染小鼠模型,给药6天后,计算小鼠体质量变化、肺指数和肺指数抑制率,苏木素—伊红(hemotaxylin-eosin staining,HE)染色观察肺组织病理变化;生化检测小鼠肺组织糖酵解代谢物丙酮酸、乳酸的含量,以及糖酵解关键酶己糖激酶(hexokinase,HK)、乳酸脱氢酶(lactate dehydrogenase,LDH)的活性;ELISA法检测小鼠肺组织炎症因子肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白细胞介素-1β(interleukin 1 Beta,IL-1β),以及抑炎因子白细胞介素-10(interleukin 10,IL-10)的变化;免疫蛋白印迹法检测肺组织中葡萄糖转运蛋白1(glucose transporter 1,GLUT1)、调控糖酵解关键因子缺氧诱导因子-1α(hypoxia inducible factor 1-Alpha,HIF-1α)、促炎巨噬细胞标记物白细胞分化抗原80(cluster of differentiation 80,CD80),以及抑炎巨噬细胞标记物白细胞分化抗原206(cluster of differentiation 206,CD206)的表达变化。结果给药6天后,与正常组相比,模型组体重明显降低(P<0.01)、肺指数显著升高(P<0.01),肺部炎性细胞浸润明显,肺组织糖酵解代谢产物丙酮酸与乳酸的含量明显升高(P<0.01)、糖酵解关键酶己糖激酶、乳酸脱氢酶的活性增强(P<0.01),肺组织炎症因子TNF-α、IL-1β水平升高(P<0.01)、抑炎因子IL-10水平降低(P<0.01),葡萄糖转运蛋白GLUT1表达降低(P<0.01)、调控糖酵解关键因子HIF-1α及促炎巨噬细胞标记物CD80呈现高表达(P<0.01),抑炎巨噬细胞标记物CD206表达减少(P<0.01)。麻杏石甘汤、奥司他韦、2-脱氧葡萄糖(2-Deoxy-D-Glucose,2DG)对以上结果均有不同程度干预。结论麻杏石甘汤可能通过抑制肺组织糖酵解过程以及糖酵解驱动的促炎巨噬细胞活化,从而抑制肺组织过度炎症反应,减轻病毒性肺炎肺损伤。
Objective To explore the mechanism of Maxing Shigan decoction in the treatment of viral pneumonia by affecting lung tissue glycolysis.Methods A mouse model infected with influenza A(H1N1)virus strain A/PR/8/34 was established by nasal inoculation.After 6 days of administration,the changes of body mass,lung index and lung index inhibition rate were calculated,and the pathological changes of lung tissues were observed by HE staining.The contents of pyruvate and lactic acid,as well as the activities of key glycolytic enzymes hexokinase and lactate dehydrogenase in lung tissues of mice were detected.The levels of inflammatory factors TNF-α,IL-1β and anti-inflammatory factor IL-10 in lung tissues of mice were detected by ELISA.The expression of glucose transporter GLUT1,HIF-1α,pro-inflammatory macrophage marker CD80,and anti-inflammatory macrophage marker CD206 in lung tissues were detected by Western blot.Results Compared with the normal group,the body weight of the model group was significantly decreased(P<0.01),the lung index was significantly increased(P<0.01),and the pulmonary inflammatory cell infiltration was obvious.The contents of pyruvate and lactic acid,which were metabolites of glycolysis,were significantly increased in lung tissues(P<0.01),the activities of key glycolysis enzymes hexokinase and lactate dehydrogenase were increased(P<0.01),the levels of inflammatory factors TNF-α and IL-1β in lung tissues were increased(P<0.01),and the levels of anti-inflammatory factor IL-10 were decreased(P<0.01).The expression of glucose transporter GLUT1 was decreased(P<0.01),HIF-1α,a key factor regulating glycolysis,and CD80,a pro-inflammatory macrophage marker,were highly expressed(P<0.01),and CD206,an anti-inflammatory macrophage marker,was decreased(P<0.01).Maxing Shigan decoction,oseltamivir and 2DG had different degrees of intervention on the above results.Conclusion Maxing Shigan decoction may inhibit excessive lung inflammation and reduce lung injury caused by viral pneumonia by inhibiting the glycolysis
作者
朱敬儒
白辰
王洋
苏晨
王旭堃
孙炜喆
卓丽清
刘铁钢
ZHU Jingru;BAI Chen;WANG Yang;SU Chen;WANG Xukun;SUN Weizhe;ZHUO Liqing;LIU Tiegang(College of traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《环球中医药》
CAS
2023年第4期620-626,共7页
Global Traditional Chinese Medicine
基金
国家中医药管理局中医药创新团队及人才支持计划(ZYYCXTD-C-202006)
国家自然科学基金(82104567)
北京中医药大学揭榜挂帅课题(2022-JYB-JBZR-030)。
关键词
麻杏石甘汤
病毒性肺炎
糖酵解
能量代谢
缺氧诱导因子
Maxing Shigan Decoction
Viral pneumonia
Glycolysis
Energy metabolism
Hypoxia inducible factor 1-Alpha