Chang'an Ⅱ Decoction(肠安Ⅱ号方)-Containing Serum Ameliorates Tumor Necrosis Factor-α-Induced Intestinal Epithelial Barrier Dysfunction via MLCK-MLC Signaling Pathway in Rats  

作　　者：CHEN Ting YIN Xiao-Ian KANG Nan WANG Xiao-ge LI Bao-shuang JI Hai-jie ZHANG Yin-qiang BIAN Li-qun ZHANG Bei-hua WANG Feng-yun TANG Xu-dong 

机构地区：[1]Department of Gastroenterology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing(100091),China [2]Graduate School of China Academy of Chinese Medical Sciences,Beijing(100700),China [3]Department of Internal Medicine,Jining Medical College Affiliated Hospital,Jining(272029),Shandong Provinee,China [4]Department of Gastroenterology,Henan University of Chinese Medicine Affiliated Hospital,Zhengzhou(136300),China [5]Department of Liver Diseases,Xiyuan Hospital,China Academy of Chinese

出　　处：《Chinese Journal of Integrative Medicine》2020年第10期745-753,共9页中国结合医学杂志（英文版）

基　　金：Supported by the National Nature Science Foundation of China(No.81373580,81173209,81704070);Visiting Scientist Program of China Academy of Chinese Medical Sciences(No.ZZ070801);"Ten Diseases and Ten Drugs"Program of Beijing Municipal Science and Technology Commission(No.Z161100000116046)。

摘　　要：Objective To investigate the effect of Chang’an Ⅱ Decoction(肠安Ⅱ号方))-containing serum on intestinal epithelial barrier dysfunction in rats.Methods Tumor necrosis factor(TNF)-α-induced injury of Caco-2 monolayers were established as an inflammatory model of human intestinal epithelium.Caco-2 monolayers were treated with blank serum and Chang’an Ⅱ Decoction-containing serum that obtained from the rats which were treated with distilled water and Chang’an Ⅱ Decoction intragastrically at doses of 0.49,0.98,1.96 g/(kg·d)for 1 week,respectively.After preparation of containing serum,cells were divided into the normal group,the model group,the Chang’an Ⅱ-H,M,and L groups(treated with 30 ng/mL TNF-αand medium plus 10%high,middle-,and low-doses Chang’an Ⅱ serum,respectively).Epithelial barrier function was assessed by transepithelial electrical resistance(TER)and permeability of fluorescein isothiocyanate(FITC)-labeled dextran.Transmission electron microscopy was used to observe the ultrastructure of tight junctions(TJs).Immunofluorescence of zonula occludens-1(ZO-1),claudin-1 and nuclear transcription factor-kappa p65(NF-κBp65)were measured to determine the protein distribution.The mRNA expression of myosin light chain kinase(MLCK)was measured by real-time polymerase chain reaction.The expression levels of MLCK,myosin light chain(MLC)and p-MLC were determined by Western blot.Results Chang’an Ⅱ Decoction-containing serum significantly attenuated the TER and paracellular permeability induced by TNF-α.It alleviated TNF-α-induced morphological alterations in TJ proteins.The increases in MLCK mRNA and MLCK,MLC and p-MLC protein expressions induced by TNF-αwere significantly inhibited in the Chang’an Ⅱ-H group.Additionally,Chang’an Ⅱ Decoction significantly attenuated translocation of NF-κBp65 into the nucleus.Conclusion High-dose Chang’an Ⅱ-containing serum attenuates TNF-α-induced intestinal barrier dysfunction.The underlying mechanism may be involved in inhibiting the MLCK

关 键 词：myosin light chain kinase-myosin light chain signaling pathway intestinal epithelial cells tight junction tumor necrosis factor-α Chang’an II Decoction drug-containing serum 

分 类 号：R285.5[医药卫生—中药学]