半夏泻心汤及其不同拆方对抗生素诱导的菌群紊乱幼鼠结肠黏膜屏障的影响

Effects of Banxia Xiexin Decoction and Its Different Disassembled Prescriptions on Colonic Mucosal Barrier in Juvenile Mice with Antibiotic-induced Microbiota Dysbiosis

目的观察半夏泻心汤及其不同拆方对抗生素诱导的菌群紊乱幼鼠结肠黏膜屏障的影响。方法70只SPF级雄性Balb/c幼鼠随机抽取10只作为空白组,其余幼鼠予混合抗生素灌胃诱导肠道菌群紊乱,连续灌胃14 d,将造模幼鼠随机分为模型组、阳性对照组、全方组、辛开组、苦降组及甘补组,每组10只。阳性对照组予双歧杆菌四联活菌片溶液0.68 g/kg灌胃,全方组、辛开组、苦降组及甘补组分别予9.1、3.19、1.82、4.1 g/kg半夏泻心汤全方及相应拆方组药液灌胃,空白组、模型组灌胃等体积生理盐水,连续14 d。末次灌胃后收集幼鼠粪便,采用16S rDNA测序分析肠道菌群结构。腹腔注射脂多糖(10 mg/kg)诱导炎症反应,ELISA检测粪便清蛋白含量,HE染色观察结肠黏膜形态,Western blot检测结肠组织紧密连接蛋白ZO-1和Occludin表达。结果与空白组比较,模型组幼鼠肠道菌群结构明显改变,结肠绒毛结构发育落后、隐窝不明显、腺体萎缩,粪便清蛋白含量明显增加(P<0.01),Occludin和ZO-1蛋白表达明显降低(P<0.01);与模型组比较,全方组和不同拆方组肠道菌群结构改善,有益菌丰度增加,有害菌丰度减少,结肠组织形态改善,粪便清蛋白含量降低,Occludin和ZO-1蛋白表达升高,总体以全方组较为显著(P<0.05,P<0.01)。结论半夏泻心汤及其不同拆方能调节抗生素诱导的菌群紊乱幼鼠肠道菌群结构,保护结肠黏膜屏障,其整体效应以半夏泻心汤全方较为明显。

Objective To observe the effects of Banxia Xiexin Decoction and its different disassembled prescriptions on colonic mucosal barrier in juvenile mice with antibiotic-induced microbiota dysbiosis.Methods 10 out of 70 SPF male Balb/c juvenile mice were randomly selected as the blank group,the others were given mixed antibiotics by gavage to induce intestinal microbiota dysbiosis.After 14 days,the modeling juvenile mice were randomly divided into the model group,positive group,Quanfang group,Xinkai group,Kujiang group and Ganbu group,with 10 mice in each group.The positive group was given Bifidobacterium tetralogy viable tablets solution by gavage of 0.68 g/kg.The Quanfang group,Xinkai group,Kujiang group and Ganbu group were given Banxia Xiexin Decoction whole prescription and corresponding disassembled prescriptions of 9.1,3.19,1.82 and 4.1 g/kg by gavage,the blank group and model group were given equal volumes of normal saline by gavage for 14 days.Feces were collected after the last gavage,the structure of intestinal microbiota was analyzed by 16S rDNA sequencing.Lipopolysaccharide was injected intraperitoneally(10 mg/kg)to induce inflammatory response.The content of fecal albumin was detected by ELISA.The morphological structure of colonic mucosa was observed by HE staining.The proteins expression of ZO-1 and Occludin in colonic tissue were detected by Western blot.Results Compared with the blank group,the composition of intestinal flora of the juvenile mice in model group changed significantly.The morphology of colon showed backward development of colonic villus structure,inconspicuous recess and gland atrophy,the content of fecal albumin increased significantly(P<0.01),and the expressions of ZO-1 and Occludin decreased significantly(P<0.01).Compared with the model group,the Quanfang group and different disassembled prescription groups adjusted the intestinal flora,increased the abundance of beneficial bacteria,and reduced the abundance of harmful bacteria,improved the tissue morphology of colon,reduced the content of fecal albumin,increased the proteins expression of ZO-1 and Occludin,which was most significant in the Quanfang group(P<0.05,P<0.01).Conclusion Banxia Xiexin Decoction and its different disassembled prescriptions can adjust the intestinal flora of juvenile mice exposed to antibiotics and protect the colonic mucosal barrier after intestinal microflora dysbiosis.The whole prescription of Banxia Xiexin Decoction is with the best efficacy.