麻黄-大黄药对抑制肺泡巨噬细胞M1极化防治急性肺损伤的作用机制

Mechanism of Mahuang-Dahuang herb pair inhibiting M1 polarization of alveolar macrophages in prevention and treatment of acute lung injury

目的探讨麻黄-大黄(Mahuang-Dahuang,MD)药对抑制肺泡巨噬细胞M1极化,减轻炎症损伤,治疗急性肺损伤的作用和机制。方法Wistar大鼠分为正常对照组(NC)、模型组(MC)、MD高(MD-H)、中(MD-M)、低剂量组(MD-L)阳性药对照组(DXMS),脂多糖腹腔注射制备急性肺损伤模型,造模后给予MD灌胃,观察肺组织病理变化;免疫组织化学法检测肺泡巨噬细胞标记物F4/80表达,F4/80和CD80、CD80和IL-6的共表达;流式细胞术检测肺组织F4/80和CD80含量的变化;PCR检测肺组织中IL-6、INOS、TNF-αmRNA相对表达;Western blot检测肺组织中CCR2、CCL2、p-p65、p65、p-p38MAPK、p38MAPK蛋白表达。结果与NC组比较,MC组大鼠肺泡结构破坏,肺间质水肿、增厚、炎细胞浸润,其中巨噬细胞数量明显增多,CCR2、CCL2蛋白表达增强,肺组织M1肺泡巨噬细胞数量增多、表达增强,炎症因子IL-6、INOS、TNF-αmRNA相对表达上调,p-p65/p65,p-p38MAPK/p38MAPK蛋白表达上调;与MC组比较,MD各组肺组织病理状态明显改善,M1肺泡巨噬细胞数量减少,表达减弱,炎症因子IL-6、INOS、TNF-αmRNA相对表达下调,CCR2、CCL2、p-p65/p65、p-p38MAPK/p38MAPK蛋白表达下调。结论MD可通过调控NF-κB和MAPK信号通路调节肺泡巨噬细胞M1极化,减少炎症因子激活和释放,抑制炎症反应,防治急性肺损伤。

Aim To investigate the effect of Mahuang-Dahuang(MD)on inhibiting M1 polarization of alveolar macrophages,alleviating inflammatory injury and treating acute lung injury,and the underlying mechanism.Methods Abandoned rats were divided into normal control group(NC),model control group(MC),MD high-dose group(MD-H),medium-dose group(MD-M),low-dose group(MD-L),and DXMS group.Lipopolysaccharide(LPS)was injected intraperitoneally to prepare an acute lung injury(ALI)rat model.After the model was established,different doses of MD were administered by intragastric administration,and the pathological changes of lung tissues were observed;immunohistochemical method was used to detect the expression of alveolar macrophage marker F4/80,F4/80 and CD80,CD80 and IL-6 Co-expression;Flow cytometry was used to detect the content of F4/80 and CD80 in lung tissues;PCR was used to detect the relative expression of IL-6,INOS,TNF-αmRNA in lung tissues;Western blot was used to detect lung tissue CCR2,CCL2,p-p65,P65,p-p38 MAPK,p38 MAPK protein expression.Results Compared with NC group,rats in MC group had damaged alveolar structure,thickened pulmonary interstitial edema,and infiltrated a large number of inflammatory cells.Among them,the number of macrophages increased significantly,which promoted the increase in the expression of macrophages-related chemokines CCR2 and CCL2.The number and expression of M1 alveolar macrophages in lung tissues increased,the relative expression of M1 alveolar macrophages-related cytokines IL-6,INOS and TNF-αmRNA was up-regulated,and the target protein in the related pathways of alveolar macrophages M1 polarization p-NF-κBp65/NF-κBp65,p-P38MAPK/p38MAPK protein expression was up-regulated.Compared with MC group,the pathological status of lung tissues in MD high,medium,and low dose groups was significantly improved,the number of lung tissue M1alveolar macrophages decreased,the relative expression of inflammatory factors IL-6,INOS and TNF-αmRNA was down-regulated,and the protein expression of CCR2,CCL2,p-NF-κBp65/NF-κBp65 and pP38MAPK/p38MAPK was reduced in lung tissues.Conclusions MD can inhibit the M1 polarization of alveolar macrophages,reduce the activation and release of inflammatory factors,inhibit inflammatory response,and prevent acute lung injury by regulating the expression of related indicators of NF-κB and MAPK signaling pathways.