加减大柴胡汤对胆囊胆固醇结石湿热证模型小鼠回肠组织病理及肠道菌群的影响  

Effect of Modified Dachaihu Decoction(加减大柴胡汤)on Ileum Histopathology and Intestinal Flora in Cholesterol Gallstones Mice with Damp-heat Syndrome

作　　者：闵莉[1] 王素英 林雪娟[1] 丁珊珊[1] 朱龙[1] 陈梅妹[1] 谢玉春 MIN Li;WANG Suying;LIN Xuejuan;DING Shanshan;ZHU Long;CHEN Meimei;XIE Yuchun(Fujian University of Traditional Chinese Medicine,Fuzhou,350122)

机构地区：[1]福建中医药大学,福建省福州市350122

出　　处：《中医杂志》2023年第13期1375-1383,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81603541);福建省自然科学基金(2021J01928)。

摘　　要：目的探讨加减大柴胡汤对胆囊胆固醇结石湿热证模型小鼠的干预作用及可能作用机制。方法将48只小鼠随机分为空白组、模型组、加减大柴胡汤组和熊去氧胆酸组,每组12只。除空白组外,其余各组小鼠采用“内湿+外湿+高胆固醇致石饲料”建立胆囊胆固醇结石湿热证小鼠模型。造模成功后,加减大柴胡汤组小鼠给予浓度为1.17 g/ml的加减大柴胡汤药液0.02 ml/g灌胃,熊去氧胆酸组小鼠给予浓度为2.5 mg/ml的熊去氧胆酸溶液0.02 ml/g灌胃,同时空白组和模型组给予0.02 ml/g生理盐水灌胃,每日1次,共干预4周。自造模起每周观察记录各组小鼠体质量、饮食量、皮毛、二便等一般情况变化,造模前后及给药干预后均进行旷场实验(包括水平穿越格子数、直立次数和静止时间)评价小鼠活跃情况;干预结束后检测胆汁中总胆固醇(TC)、总胆汁酸(TBA)含量;HE染色法观察回肠组织病理学变化;16S rDNA扩增测序法检测各组小鼠肠道菌群生物多样性。结果干预后与空白组比较,模型组小鼠水平穿越格子数和直立次数均减少、静止时间均延长,胆汁内TC含量升高、TBA含量降低(P<0.01)。与模型组和熊去氧胆酸组比较,加减大柴胡汤组小鼠水平穿越格子数和直立次数增多、静止时间缩短,胆汁内TC含量均降低、TBA含量均升高(P<0.05或P<0.01)。HE染色显示,模型组小鼠回肠结构完整性被破坏,肠绒毛疏松、杂乱、断裂,固有层明显增厚;加减大柴胡汤组和熊去氧胆酸组小鼠较模型组肠绒毛密度有所增加,肠绒毛断裂等状况有所好转,其中加减大柴胡汤组小鼠回肠组织病理形态改善状况更佳。肠道菌群Alpha多样性分析结果示,与空白组比较,模型组小鼠肠道Chao1、Observed species、Pielou_e、Shannon、Simpson指数均降低(P<0.01);加减大柴胡汤组小鼠肠道Shannon、Pielou_e指数明显高于模型组和熊去氧胆酸组(P<0.05或P<0.01)。BetaObjective To investigate the therapeutic effect of Modified Dachaihu Decoction(加减大柴胡汤,MDD)on cholesterol gallstones(CS)mice with damp-heat syndrome and the possible mechanism.Methods Forty-eight mice were randomly divided into blank group,model group,MDD group and ursodeoxycholic acid group,with 12 mice in each group.Except for the blank group,the other three groups were administered with“internal damp+external damp+high cholesterol stone-inducing diet"to establish the CS damp-heat syndrome mouse model.After suc-cessful modeling,MDD group and ursodeoxycholic acid group were given O.02 ml/g of MDD at a concentration of 1.17 g/ml and 0.02 ml/g of ursodeoxycholic acid solution at a concentration of 2.5 mg/ml by gavage,respectively,while the model group and blank group were given 0.02 ml/g of normal saline by gavage,once daily,for a total of 4 weeks.The general changes of body mass,diet,fur,stool,urine and otherconditions of mice in each group were observed and recorded every week since the establishment of the model.Open field test including the number of crossings,the rearing times and resting time was performed to measure the activity of mice in each group before and after modeling and after treatment.After treatment,the contents of total cholesterol(TC)and total bile acid(TBA)in bile were detected;hematoxylin and eosin(HE)staining was used to observe the histopathological changes of the ileum;and 16SrDNA amplification sequencing was used to detect intestinal flora biodiversity in each group.Results Compared to the blank group,the mice in the model group had decreased number of crossings and rearing times,prolonged resting time,increased TC content in bile and decreased TBA content(P<0.01).Compared to the model group and ursodeoxycholic acid group,the MDD group had increased number of crossings and rearing times,shortened resting time,decreased TC content in bile and increased TBA content(P<0.05 or P<0.01).HE staining showed that the structural integrity of the ileum was destroyed in the model group,

关 键 词：胆囊胆固醇结石 湿热证 加减大柴胡汤 肠道菌群 旷场实验 回肠组织 

分 类 号：R285.5[医药卫生—中药学]