基于肺泡巨噬细胞M2极化的麻黄-大黄药对治疗急性肺损伤的作用机制研究  

Mechanism of Ephedrae Herba-Rhei Radix et Rhizoma drug pair on treating acute lung injury based on alveolar macrophage M2 polarization

作　　者：王旭红 闫曙光[2] 惠毅[2] 史捷 李京涛 WANG Xu-hong;YAN Shu-guang;HUI Yi;SHI Jie;LI Jing-tao(Department of Respiratory,Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China;College of Basic Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Infectious Disease,Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学附属医院呼吸科,陕西咸阳712000 [2]陕西中医药大学基础医学院,陕西咸阳712046 [3]陕西中医药大学附属医院感染科,陕西咸阳712000

出　　处：《中草药》2022年第9期2715-2722,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81703974);陕西省自然科学基础研究计划面上项目(2021JM-473);陕西中医药大学学科创新团队建设项目(2019-YL05)。

摘　　要：目的 探讨麻黄-大黄药对对急性肺损伤(acute lung injury,ALI)大鼠肺泡替代激活的巨噬细胞(alternatively activated macrophages,M2)极化的调控作用及其机制。方法 Wistar大鼠随机分为对照组、模型组及麻黄-大黄低、中、高剂量(2.2、4.4、8.8 g/kg)组和地塞米松(5 mg/kg)组,大鼠ip脂多糖(lipopolysaccharide,LPS)制备ALI模型,造模后立即给予药物进行干预,采用苏木素-伊红(HE)染色法观察肺组织病理变化;采用免疫荧光双标法检测肺泡巨噬细胞标记物F4/80和M2标记物CD206,以及F4/80和白细胞介素-10(interleukin-10,IL-10)的共表达;采用流式细胞术检测肺组织F4/80和CD206阳性巨噬细胞数量的变化;采用q RT-PCR法检测肺组织中精氨酸酶-1(arginase-1,Arg-1)和IL-10 m RNA表达;采用Western blotting法检测肺组织中Arg-1和IL-10蛋白表达。结果 与对照组比较,模型组大鼠肺组织肺泡结构破坏,肺间质水肿增厚,炎性细胞聚集、浸润,其中巨噬细胞数量显著增多;免疫组化和流式细胞术检测结果证实,肺泡巨噬细胞数量增多、表达增强,M2巨噬细胞表达增强、数量增多;M2肺泡巨噬细胞相关细胞因子IL-10和Arg-1 mRNA和蛋白表达水平均显著升高(P<0.01)。与模型组比较,麻黄-大黄高、中剂量组肺组织病理状态明显改善,肺组织M2肺泡巨噬细胞数量进一步升高,抗炎因子IL-10和促修复因子Arg-1蛋白和m RNA表达均显著上调(P<0.01)。结论 麻黄-大黄药对可促进肺泡巨噬细胞M2极化,增加抗炎因子激活和释放,抑制炎症反应,从而治疗急性肺损伤。Objective To investigate the regulatory effect and mechanism of Mahuang(Ephedrae Herba)-Dahuang(Rhei Radix et Rhizoma) drug pair(MD) on polarization of alternatively activated macrophages(M2) in rats with acute lung injury(ALI).Methods Wistar rats were randomly divided into control group,model group,MD low-,medium-and high-dose(2.2,4.4,8.8 g/kg) groups and dexamethasone(5 mg/kg) group.Lipopolysaccharide(LPS) was used to prepare ALI model,and drug intervention was given immediately after modeling,and hematoxylin-eosin(HE) staining was used to observe the pathological changes of lung tissue;Immunofluorescence double-labeling method was used to detect alveolar macrophage marker F4/80 and M2 marker CD206,as well as the co-expression of F4/80 and interleukin-10(IL-10);Flow cytometry was used to detect the changes in number of F4/80 and CD206 positive macrophages in lung tissue;Arginase-1(Arg-1) and IL-10 mRNA expressions in lung tissue were detected by qRTPCR;Arg-1 and IL-10 protein expressions in lung tissue were detected by Western blotting.Results Compared with control group,alveolar structure in lung tissue of rats in model group was destroyed,pulmonary interstitial edema was thickened,inflammatory cells were aggregated and infiltrated,and number of macrophages was significantly increased;The results of immunohistochemistry and flow cytometry confirmed that number and expression of alveolar macrophages were increased,expression and number of M2macrophages were increased;mRNA and protein expressions of M2 alveolar macrophage-related cytokines IL-10 and Arg-1 were significantly increased(P < 0.01).Compared with model group,pathological state of lung tissue in MD high-and medium-dose groups were significantly improved,number of M2 alveolar macrophages in lung tissue was further increased,protein and mRNA expressions of anti-inflammatory factor IL-10 and pro-repair factor Arg-1 were significantly up-regulated(P < 0.01).Conclusion MD can promote M2 polarization of alveolar macrophages,increase the activation and releas

关 键 词：麻黄-大黄药对 急性肺损伤 肺泡巨噬细胞 M2极化 炎症反应 

分 类 号：R285.5[医药卫生—中药学]