摘要
目的:探讨怡情止泻汤调控内质网应激/NF-κB通路治疗溃疡性结肠炎(UC)的作用机制,为UC临床治疗提供参考。方法:选取SPF级健康C57BL/6小鼠60只,按照随机数字表法将其随机分为正常对照组、模型对照组、怡情止泻汤8、16、32 g/kg组、5-氨基水杨酸1 g/kg组,每组10只,正常对照组予以生理盐水灌胃,其余5组建立肝郁脾虚型UC小鼠模型后,分别予以生理盐水或相应药物灌胃,连续7 d。观察各组小鼠一般情况,进行疾病活动指数(DAI)评分,HE染色观察肠粘膜病理损伤,进行肠粘膜病理(Geboes)评分,酶联免疫吸附法检测炎症因子水平,蛋白免疫印迹法(Western Blot)检测相关蛋白表达。结果:与正常对照组比较,模型对照组小鼠肠粘膜损伤严重,DAI评分、Geboes评分显著升高(P<0.01),结肠组织TNF-α、IL-6、IL-1β含量、ATF4蛋白表达、p-PERK/PERK、p-eIF2α/eIF2α、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα比值均明显升高(P<0.01)。与模型对照组比较,怡情止泻汤8、16、32 g/kg组、5-氨基水杨酸1 g/kg组小鼠经药物灌胃治疗后肠粘膜病理损伤、一般情况明显改善,DAI评分、Geboes评分显著降低(P<0.01),结肠组织TNF-α、IL-6、IL-1β含量显著降低,ATF4蛋白表达、p-PERK/PERK、p-eIF2α/eIF2α、p-IκBα/IκBα、p-NF-κB p65/NF-κB p65比值显著降低(P<0.01)。结论:怡情止泻汤可有效治疗肝郁脾虚型UC小鼠,减少炎症介质释放,抑制炎症反应,其治疗机制可能与抑制内质网应激诱导的NF-κB通路激活有关。
Objective:To explore the mechanism of Yiqing Zhixie Decoction in regulating endoplasmic reticulum stress/NF-κB pathway and thus provide reference for the clinical treatment of ulcerative colitis(UC).Methods:Sxity SPF-grade C57 BL/6 mice were randomly assigned into the normal control group,model group,Yiqing Zhixie Decoction(8,16,and 32 g/kg)groups,and 5-aminosalicylic acid(1 g/kg)group,with 10 mice in each group.The normal control group was given normal saline water by gavage,and the other 5 groups were given normal saline or corresponding drugs by gavage for 7 days after the establishment of UC model with Ganyupixu(肝郁脾虚)syndrome.The general conditions of mice in each group were observed,and the disease activity index(DAI)score was assessed.The pathological damage of intestinal mucosa was observed via HE staining and evaluated based on the Geboes score.The levels of inflammatory cytokines in each group were determined by enzyme-linked immunosorbent assay,and the expression levels of related proteins by Western blotting.Results:Compared with the normal control group,the modeling caused serious injury in intestinal mucosa,increased DAI score and Geboes score(P<0.01),up-regulated the ATF4 protein level,and elevated the TNF-α,IL-6,and IL-1βlevels as well as p-PERK/PERK,p-eIF2α/eIF2α,p-NF-κB p65/NF-κB p65,and p-IκBα/IκBαratios in colon tissue(P<0.01).Compared with the model control group,Yiqing Zhixie Decoction(8,16,and 32 g/kg)and 5-aminosalicylic acid(1 g/kg)alleviated the pathological injury,improved the general conditions of mice,lowered the DAI score and Geboes score(P<0.01),down-regulated the ATF4 protein level,and decreased the TNF-α,IL-6,and IL-1βlevels as well as p-PERK/PERK,p-eIF2α/eIF2α,p-NF-κB p65/NF-κB p65,and p-IκBα/IκBαratios(P<0.01).Conclusion:Yiqing Zhixie Decoction can effectively treat UC mice with Ganyupixu(肝郁脾虚)syndrome.It may reduce the release of inflammatory mediators and inhibit inflammatory reaction by blocking the endoplasmic reticulum stress-induced activation of NF-κB pathway.
作者
郑春菊
王斌
赵恩春
郑勇
Zheng Chunju;Wang Bin;Zhao Enchun;Zheng Yong(Department of Anorectal Medicine,Huai'an Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Huai'an 223001)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第2期173-178,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
淮安市自然科学基金项目(编号:HAB202039)
江苏省中医药局科技项目(编号:YB2017069)
关键词
怡情止泻汤
溃疡性结肠炎
内质网应激
核转录因子B通路
炎症反应
Yiqing Zhixie Decoction
ulcerative colitis
endoplasmic reticulum stress
nuclear factor kappa B pathway
inflammatory reaction
作者简介
通信作者:王斌,Email:229415621@qq.com;郑春菊,E-mail:zhengchunjujs_zcj@163.com。
