Effect of Shenling Baizhu Powder on Diarrhea Regulating the Interation Intestinal Flora and Epithedial Cell Junction in Diarrhea Young Rats
诸葛远莉 1申中美 2龚琳霞 1宋祖贤 1崔倪 1夏小兰1
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作者信息
1. 南京中医药大学附属南京市中西医结合医院,江苏 南京 210014
2. 江苏省苏州市吴江区中医院脑病科,江苏 苏州 215100
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摘要
目的:探讨参苓白术散对于改善幼鼠腹泻模型及调节肠道菌群-上皮细胞相互作用机制.方法:通过灌胃大黄水煎液建立幼鼠腹泻模型,参苓白术散按照临床折算剂量设 20.0、10.0 和 5.0g药材/kg剂量灌胃给药2 周.试验期间每周测定体重变化;检测指标包括:血浆D乳酸、内毒素水平;结肠组织形态和黏液状态(PAS染色);紧密连接蛋白分子(ZO-1,Occludin和Claudin)的mRNA和蛋白表达水平;肠道菌群分析(选择性培养基).结果:参苓白术散高、中剂量组在改善腹泻症状同时显著抑制体重减轻.参苓白术散高、中剂量组可下调模型组血清内毒素和 D-乳酸水平.结肠组织 PAS 染色病理学结果显示,参苓白术散各剂量组可改善模型大鼠肠道绒毛损伤,同时恢复杯状细胞和黏液量.参苓白术散高剂量组Occludin和ZO-1 表达高于模型组,Claudin 表达略微增高.参苓白术散高、中剂量组可减少盲肠内容物中肠杆菌、肠球菌数量,并使双歧杆菌、乳杆菌的数量接近正常数量.结论:参苓白术散改善模型大鼠肠道菌群失调,同时上调肠道紧密连接蛋白表达,恢复肠道上皮细胞屏障,进而改善腹泻模型肠道症状.
Abstract
Objective:To investigate the influence of Shenling Baizhu powder(SLP)on improving diar-rhea model and regulating the interaction between intestinal flora and epithelial cell in young rats.Methods:The diarrhea model of young mice was established by gavage of rhubarb water decoction.SLP was administra-ted by gavage for 2 weeks at the dose of 20.0,10.0 and 5.0 g herbs/kg.The changes of body weight were measured every week during the experiment;the levels of D-lactic acid and endotoxin in plasma were meas-ured by kit;the morphology and mucus state of colon tissue were detected by PAS staining;the mRNA and protein expression of tight junction protein(occludin,ZO-1 and claudin)were detected by real-time PCR and Western blot respectively;The intestinal flora was analyzed with selective medium.Results:SLP could improve the diarrhea symptoms and significantly inhibit the weight loss.High and medium dose of SLP signifi-cantly decreased serum D-lactic acid and endotoxin.The results of PAS staining showed that each dose group of SLP could improve the villi injury of intestinal tissue of model rats,and recover the content of goblet cells and mucus.The expression of ZO-1 and occludin were significantly up-regulated in the high-dose SLP group,and the expression of claudin was also up-regulated.High and medium dose groups of SLP could sig-nificantly reduce the number of Enterobacteriaceae and Enterococcus in cecal contents,and recover the num-ber of Lactobacillus and Bifidobacterium.Conclusion:SLP can improve the dysbiosis,increase the expres-sion of tight junction protein,restore the intestinal epithelial cell barrier,and then improve the intestinal symptoms of diarrhea model rats.
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