基于AQP3/Claudin-1研究痛泻要方治疗腹泻型肠胃激综合征模型大鼠的机制  

Study on Mechanism of Tongxie Yaofang(痛泻要方)in Treatment of IBS-D Model Rats Based on AQP3/Claudin-1

作　　者：王玲玲 柏茂树[1] 吴至久[2] WANG Lingling;BAI Maoshu;WU Zhijiu(Dazhou Hosptial of Integrated Traditional Chinese and Western Medicine,Dazhou 635000,Sichuan,China;Hospital Affiliated to North Sichuan Medical College,Nanchong 637000,Sichuan,China)

机构地区：[1]达州市中西医结合医院,四川达州635000 [2]南充市川北医学院附属医院,四川南充637000

出　　处：《辽宁中医杂志》2024年第3期200-204,I0009,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：四川省科技项目(2017JY0202);四川省中医药管理局项目(2020LC0099)。

摘　　要：目的通过构建腹泻型肠易激综合征(irritable bowel with diarrhea,IBS-D)大鼠模型,观察经痛泻要方治疗后肥大细胞(mast cell,MC)计数、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的浓度变化,水通道蛋白3(aquaporin 3,AQP3)和Claudin-1 mRNA及蛋白的表达水平,并探讨痛泻要方治疗IBS-D的作用机制。方法采用改良慢性应激结合束缚的方法建立IBS-D模型大鼠。甲苯胺蓝染色法进行MC染色计数,ELISA法检测TNF-α含量,RT-PCR及SABC免疫组化法检测结肠组织AQP3和Claudin-1mRNA及蛋白的表达水平。结果(1)与正常组比较,模型组MC计数增加,差异有统计学意义(P<0.05);与模型组比较,匹维溴铵组、痛泻要方中、高剂量组MC计数明显减少,差异有统计学意义(P<0.05);(2)与正常组比较,模型组TNF-α含量明显上升,差异有统计学意义(P<0.05);与模型组比较,匹维溴铵组、痛泻要方中、高剂量组TNF-α含量均下降,差异有统计学意义(P<0.05);(3)与正常组比较,模型组AQP3和Claudin-1mRNA及蛋白表达水平降低,差异有统计学意义(P<0.05);与模型组比较,痛泻要方中、高剂量组及匹维溴铵组能上调大鼠结肠AQP3及Claudin-1mRNA及蛋白的表达水平,差异有统计学意义(P<0.05)。结论痛泻要方可能通过降低MC计数,降低TNF-α含量,上调AQP3和Claudin-1的表达水平,促进结肠对肠腔内水分的重吸收,修复肠道黏膜屏障,对IBS-D发挥作用。Objective To describe the amounts of mast cell(MC)and the changes of tumor necrosis factor-α(TNF-α)concentration,the expression of aquaporin 3(AQP3)and Claudin-1 and investigate the mechanism of Tongxie Yaofang(痛泻要方,TXYF)in the treatment of diarrhea-induced irritable bowel syndrome(IBS-D)model rats.Methods IBS-D model was made by modified method of chronic restraint stress combined with restraint method.MC staining was performed using toluidine blue staining.TNF-αcontent was detected using ELISA and the expression levels of AQP3 and Claudin-1 mRNA and protein in colon tissue were detected using RT-PCR and SABC immunohistochemistry.Results(1)Compared with the normal group,the model group showed an increase in MC count,with a statistically significant difference(P<0.05).Compared with the model group,the MC count in the pivoxil bromide group,the TXYF middle and high dose groups significantly decreased,with a statistically significant difference(P<0.05).(2)Compared with that of the normal group,the TNF-αcontent in the model group was significantly increased,with a statistically significant difference(P<0.05).Compared with those of the model group,the levels of TNF-αin the pivoxil bromide group,the TXYF middle and high-dose group were all decreased,and the difference was statistically significant(P<0.05).(3)Compared with those of the normal group,the mRNA and protein expression levels of AQP3 and Claudin-1 in the model group were decreased significantly(P<0.05).Compared with the model group,the TXYF middle and high dose groups and the pivovir bromide group were able to up-regulate the expression levels of AQP3 and Claudin-1 mRNA and protein in the colon of rats,with statistically significant differences(P<0.05).Conclusion TXYF may play a role in IBS-D by reducing MC count,reducing TNF-αcontent,increasing the expressions of AQP3 and Claudin-1,promoting the reabsorption of water in the intestinal cavity by the colon and repairing the intestinal mucosal barrier.

关 键 词：痛泻要方 腹泻型肠易激综合征 水通道蛋白3 紧密连接蛋白1 

分 类 号：R574[医药卫生—消化系统]