Objective:To investigate the protective effect of Chinese herbal formula Huangqin Decoction(HQD)on ulcerative colitis mouse model induced by dextran sulphate sodium(DSS)and human intestinal epithelial cell injury indu...Objective:To investigate the protective effect of Chinese herbal formula Huangqin Decoction(HQD)on ulcerative colitis mouse model induced by dextran sulphate sodium(DSS)and human intestinal epithelial cell injury induced by tumour necrosis factor-α(TNF-α).Methods:In vivo,30 male C57BL/6 mice were divided into 5 groups using a random number table(n=6 per group),including control,DSS,5-aminosalicylic acid(5-ASA),HQD low-(HQD-L)and high-dose(HQD-H)groups.The colitis mouse model was established by 3%(w/v)DSS water for 5 days.Meanwhile,mice in the HQD-L,HQD-H and 5-ASA groups were administrated with 100,200 mg/kg HQD or 100 mg/kg 5-ASA,respectively,once daily by gavage.After 9 days of administration,the body weight,disease activity index(DAI)score and colon length of mice were measured,the pathological changes of colons were analyzed by hematoxylin-eosin staining(HE)staining,and the levels of serum interleukin(IL)-6,IL-1βand TNF-αwere measured by enzyme linked immunosorbent assay.In vitro,the human colon epithelial normal cells(FHC cells)were exposed to HQD(0.6 mg/mL)for 12 h and then treated with TNF-α(10 ng/mL)for 24 h.The tight junction(TJ)protein expression levels of Claudin-4 and Occludin,and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB(NF-κB)-α(IκBα)were measured by Western blot.Results:In vivo,compared with the DSS group,HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day(P<0.05).Moreover,HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice(P<0.05).HE staining showed HQD attenuated the pathological changes of colitis mice,and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group(P<0.05).Meanwhile,HQD-H and 5-ASA significantly decreased the serum IL-1β,IL-6 and TNF-αlevels of mice(P<0.05).In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBαlevels in FHC cells compared with the TNF-αgroup(P<0.05).Conclusion:HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.展开更多
基金the Scientific Research Project of Jiangsu Provincial Administration of Traditional Chinese Medicine(No.JD2019SZXYB05)Natural Science Foundation of Nanjing University of Chinese Medicine(No.XZR2020030)National Administration of Traditional Chinese Medicine:2019 Project of Building Evidence-Based Practice Capacity for Traditional Chinese Medicine(No.2019XZZX-XH007)。
文摘Objective:To investigate the protective effect of Chinese herbal formula Huangqin Decoction(HQD)on ulcerative colitis mouse model induced by dextran sulphate sodium(DSS)and human intestinal epithelial cell injury induced by tumour necrosis factor-α(TNF-α).Methods:In vivo,30 male C57BL/6 mice were divided into 5 groups using a random number table(n=6 per group),including control,DSS,5-aminosalicylic acid(5-ASA),HQD low-(HQD-L)and high-dose(HQD-H)groups.The colitis mouse model was established by 3%(w/v)DSS water for 5 days.Meanwhile,mice in the HQD-L,HQD-H and 5-ASA groups were administrated with 100,200 mg/kg HQD or 100 mg/kg 5-ASA,respectively,once daily by gavage.After 9 days of administration,the body weight,disease activity index(DAI)score and colon length of mice were measured,the pathological changes of colons were analyzed by hematoxylin-eosin staining(HE)staining,and the levels of serum interleukin(IL)-6,IL-1βand TNF-αwere measured by enzyme linked immunosorbent assay.In vitro,the human colon epithelial normal cells(FHC cells)were exposed to HQD(0.6 mg/mL)for 12 h and then treated with TNF-α(10 ng/mL)for 24 h.The tight junction(TJ)protein expression levels of Claudin-4 and Occludin,and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB(NF-κB)-α(IκBα)were measured by Western blot.Results:In vivo,compared with the DSS group,HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day(P<0.05).Moreover,HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice(P<0.05).HE staining showed HQD attenuated the pathological changes of colitis mice,and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group(P<0.05).Meanwhile,HQD-H and 5-ASA significantly decreased the serum IL-1β,IL-6 and TNF-αlevels of mice(P<0.05).In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBαlevels in FHC cells compared with the TNF-αgroup(P<0.05).Conclusion:HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.
