参芪颗粒对脂多糖诱导的小鼠急性肺损伤的作用及机制研究  

Effect and Mechanism of Shenqi(参芪)Granules in Preventing Acute Lung Injury Induced by Lipopolysaccharide in Mice

作　　者：杨薇[1] 许洪玲 吴瑕[1] 袁崇均[1] 陈帅[1] 罗森[1] 袁明铭 王媛 张磊[1,2,3] 周静 YANG Wei;XU Hongling;WU Xia;YUAN Chongjun;CHEN Shuai;LUO Sen;YUAN Mingming;WANG Yuan;ZHANG Lei;ZHOU Jing(Sichuan Academy of Chinese Medicine Sciences,Chengdu 610041;School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137;Sichuan-Chongqing Joint Key Laboratory of Innovation of New Drugs of Traditional Chinese Medicine,Chengdu 610041)

机构地区：[1]四川省中医药科学院,成都610041 [2]成都中医药大学药学院,成都611137 [3]中药新药创制川渝共建重点实验室,成都610041

出　　处：《中药药理与临床》2024年第9期30-36,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省科技计划项目(编号:2022YFS0432);四川省中医药管理局科学技术研究专项(编号:2021ZD002、2024ZD01)。

摘　　要：目的:观察参芪颗粒对脂多糖(LPS)诱导的小鼠急性肺损伤的预防作用及可能机制。方法:小鼠单核巨噬细胞RAW264.7加入100μg/mL LPS刺激6 h造成细胞炎症模型,随机分为空白对照组、模型对照组、地塞米松20μg/mL组、参芪颗粒32.5、65、130μg/mL组,采用CCK-8法检测细胞存活率;采用流式细胞仪检测细胞线粒体膜电位(JC-1)的变化。C57BL/6小鼠随机分为正常对照组、模型对照组、参芪颗粒含生药7、14、28 g/kg组、地塞米松50 mg/kg组,各组分别灌胃给予相应药物或生理盐水,1次/d;给药7 d后,除正常对照组外,其余各组动物气管滴注20 mg/kg LPS诱导小鼠急性肺损伤模型。造模24 h后检测小鼠肺湿质量/干质量比以评价肺水肿程度;酶联免疫吸附法测定小鼠肺泡灌洗液和血清中的肿瘤坏死因子α(TNF-α)、白介素6(IL-6)、IL-1β、一氧化氮(NO)含量;HE染色法检测肺组织病理损伤情况;流式细胞术测定小鼠外周血中的T、B淋巴细胞及脾脏组织中的NK细胞。结果:与空白对照组比较,模型对照组细胞呈多边形不规则生长,突触明显,细胞存活率显著增加,细胞线粒体膜电位则显著降低(P<0.01);与模型对照组比较,参芪颗粒65、130 mg/mL组能明显升高小鼠单核巨噬细胞RAW264.7细胞线粒体膜电位(P<0.05或P<0.01),并显著降低LPS诱导的巨噬细胞活性升高(P<0.01)。与正常对照组比较,模型对照组小鼠肺损伤评分显著升高,可见支气管内炎性细胞浸润,肺间质纤维增生等,肺组织湿/干质量比、外周血中总CD3^(+)T细胞、CD3^(+)CD4^(+)T细胞、B淋巴细胞百分率、脾脏中NK细胞百分率明显降低,血清与肺灌洗液中的TNF-α、IL-1β、IL-6和NO的含量明显升高(P<0.05或P<0.01);与模型对照组比较,参芪颗粒28 g/kg组能较好地保护肺脏组织的病理损伤,降低肺损伤评分,缓解肺水肿程度,升高外周血中总CD3^(+)T细胞、B细胞及脾脏组织中NK细胞的百分率,各给Objective:To observe the effect and possible mechanism of Shenqi(参芪)granules in preventing lipopolysaccharide(LPS)-induced acute lung injury in mice.Methods:RAW264.7 cells were stimulated with 100μg/mL LPS for 6 h for the modeling of cellular inflammation.Cells were randomized into normal control,model control,dexamethasone(20μg/mL),and Shenqi granules(32.5,65,and 130μg/mL)groups.The cell viability and mitochondrial membrane potential(JC-1)were measured by the CCK-8 assay and flow cytometry,respectively.C57BL/6 mice were randomized into normal control,model control,Shenqi granules(28,14,and 7 g/kg),and dexamethasone(50 mg/kg)groups.After 7 days of administration with corresponding drugs or normal saline once a day,other groups except the normal control group were subjected to intratracheal instillation of 20 mg/kg LPS for the modeling of acute lung injury in mice.After 24 h,the wet-dry weight(W/D)ratio of mouse lungs was calculated to evaluate lung edema.Enzyme-linked immunosorbent assay was employed to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),and nitric oxide(NO)in the bronchoalveolar lavage fluid and serum.Hematoxylin-eosin staining was used to observe the pathological damage in the lung tissue.Flow cytometry was employed to determine the percentages of T and B lymphocytes in the peripheral blood and the natural killer(NK)cells in the splenic tissue.Results:Compared with the normal control group,the model control group showed polygonal irregular growth,protrusions,increased survival rate,and decreased mitochondrial membrane potential of RAW264.7 cells(P<0.01).Compared with the model control group,Shenqi granules(65,and 130μg/mL)increased the mitochondrial membrane potential of RAW264.7 cells(P<0.05 or P<0.01)and attenuated the cell viability increase caused by LPS.Compared with the normal control group,the model control of mice presented increased lung injury scores,endobronchial inflammatory cell infiltration,and interstitial fibroplasia.The m

关 键 词：参芪颗粒 急性肺损伤 炎症反应 线粒体膜电位 巨噬细胞 免疫 

分 类 号：R285.5[医药卫生—中药学]