大黄调节肠道菌群紊乱发挥保护急性肺损伤的作用机制研究  

Rhubarb protects mice from acute lung injury induced by lipopolysaccharide through improving intestinal microbial disorders

作　　者：李志军[1] 汪飞[1] 刘娟 叶芃 LI Zhijun;WANG Fei;LIU Juan;YE Peng(Department of Respiratory,Zhejiang Hospital,Hangzhou 310012,China)

机构地区：[1]浙江医院呼吸内科,杭州310012

出　　处：《浙江医学》2023年第12期1249-1254,I0005,I0006,共8页Zhejiang Medical Journal

基　　金：国家自然科学基金项目(81973590);浙江省基础公益研究计划项目(LY18H290008)。

摘　　要：目的探讨大黄通过调节肠道菌群紊乱而发挥保护急性肺损伤的作用机制。方法采用随机抽签法将36只小鼠分为正常对照组、急性肺损伤组、曲古抑菌素A组、丙戊酸组、高剂量大黄组和低剂量大黄组,每组6只。除正常对照组外,其余各组均采用气管内滴注脂多糖(LPS)建立急性肺损伤动物模型。曲古抑菌素A组:腹腔注射25 ml/kg曲古抑菌素A;丙戊酸组:腹腔注射25 ml/kg丙戊酸;高剂量大黄组:胃部灌注150 mg/kg大黄流浸膏;低剂量大黄组:胃部灌注50 mg/kg大黄流浸膏;正常对照组及急性肺损伤组腹腔注射等体积0.9%氯化钠溶液(25 ml/kg)。小鼠处死后分离肺组织进行HE染色和肺组织湿/干(W/D)比值评估肺组织损伤程度,采用ELISA法检测肺组织和血清中组蛋白乙酰化酶(HDAC)6及HDAC9水平;采用流式细胞仪检测脾组织中Th17/调节性T淋巴细胞(Treg);收集小鼠粪便样本并采用细菌16S核糖体DNA高通量测序技术检测肠道菌群构成及丰度。结果LPS诱导急性肺损伤动物模型伴随肺泡间隔增厚和中性粒细胞浸润,肺组织W/D比值升高,采用大黄干预后能明显减轻肺组织损伤,降低肺组织W/D比值。与正常对照组比较,高剂量大黄组小鼠血清HDAC6水平升高(P<0.05)。与正常对照组比较,急性肺损伤组脾脏组织中Treg数量增加,Th17数量降低,差异均有统计学意义(均P<0.05)。大黄处理后能恢复Th17/Treg细胞失衡。此外,急性肺损伤小鼠呈现厚壁菌门/拟杆菌门菌群比例升高,表现为厚壁菌门减少,拟杆菌门增加。大黄干预后,在属水平上增加了阿利斯蒂普菌、梭状杆菌和乳酸杆菌的增殖。结论大黄通过改善急性肺损伤肠道菌群紊乱,提升血清中HDAC6水平,恢复Th17/Treg细胞平衡,从而发挥保护急性肺损伤的作用。Objective To investigate the protective effect of rhubarb on acute lung injury induced by lipopolysaccharide(LPS)and its mechanism in mice.Methods Thirty-six male C57BL/6 mice were randomly divided into 6 groups:normal control group,acute lung injury group,trichostatin A group,valproic acid group,high-dose rhubarb group,and lowdose rhubarb group with 6 mice in each group.Intratracheal instillation of lipopolysaccharide was used to induce acute lung injury model.The mice were intervened with trichostatin A(25 ml/kg),valproic acid(25 ml/kg),high-dose rhubarb(150 mg/kg,gavage)or low-dose rhubarb(50 mg/kg,gavage),respectively.The equal volume of saline was injected intraperitoneally in normal control group and acute lung injury group.After the mice were euthanized,lung tissue was harvested for HE staining and wet/dry(W/D)ratio;and the concentrations of histone deacetylase(HDAC)6 and HDAC9 in lung tissue and peripheral serum were detected using ELISA,the proportion of helper T lymphocytes/regulatory T cells lymphocytes(Th17/Treg)was tested in splenic tissue by flow cytometry,fecal samples was collected for determining the composition and abundance of intestinal microbiota by 16S rDNA high-throughput sequencing.Results Mice in the acute lung injury group showed the thickening of alveolar septa,infiltration of neutrophils,and increased lung W/D ratio.Rhubarb alleviated lung tissue damage and decreased the ratio of W/D.Compared with the normal control group,the serum HDAC6 level in the high-dose rhubarb group was increased(P<0.05).Compared with the normal control group,the number of Treg cells increased in the spleen tissue of the acute lung injury group,and the number of Th17 cells decreased(both P<0.05).Th17/Treg cell imbalance was restored after treatment by rhubarb.The proportion of Firmicutes/Bacteroidetes was increased,with an increased Firmicutes and a decreased Bacteroidetes.The rhubarb intervention promoted the proliferation of Alisteptococcus,Clostridium,and Lactobacillus at the genus level.Conclusion Rhubarb p

关 键 词：大黄 急性肺损伤 肠道微生物群 辅助性T淋巴细胞/调节性T淋巴细胞 组蛋白乙酰化酶 

分 类 号：R285.5[医药卫生—中药学]