摘要
目的观察痛泻要方对腹泻型肠易激综合征(Irritable bowel syndrome with diarrhea,IBS-D)模型大鼠法尼醇受体(Farnesol receptor,FXR)、调节核因子-κB(Nuclear factor-κB,NF-κB)、炎性因子的影响,探讨痛泻要方干预IBS-D的抗炎机制。方法健康SD大鼠随机分为空白组(Control组)、模型组(Model组)、痛泻要方低剂量组(TXYF-L组,4.977 g/kg)、痛泻要方中剂量组(TXYF-M组,9.954 g/kg)、痛泻要方高剂量组(TXYF-H组,19.908 g/kg)、匹维溴铵组(PWXA组,0.018 g/kg),每组10只。除空白组外,其余组均采用采用慢性束缚应激+番泻叶灌胃建立肝郁脾虚型IBS-D大鼠模型。模型建立成功后,按相应组别处理方法连续干预14 d。观察各组大鼠干预后的一般情况、腹泻率、粪便含水率,腹壁回缩反射达到3分时的注水量、糖水偏嗜率、血清D-木糖水平,HE染色观察大鼠结肠组织形态结构变化,PCR法检测结肠组织中FXR基因表达,Western blot方法检测结肠组织中FXR蛋白、NF-κB蛋白表达,ELISA检测血清IL-6、IL-8、TNF-α水平。结果经痛泻要方治疗后,大鼠体重增加,粪便含水率降低,糖水偏嗜率升高,大鼠达到腹部回缩反射评分3分时的注水量增加,血清中D-木糖含量升高、摄食量增加。HE染色可见Model组黏膜上皮出现绒毛状或者指状的结构,黏膜上层中有炎细胞浸润。经治疗后,TXYF-M组黏膜上皮平整,绒毛状或者指状结构消失,炎细胞浸润程度降低。痛泻要方高剂量组大鼠结肠组织中FXR mRNA及FXR蛋白表达明显升高,血清中IL-6、IL-8、TNF-α浓度及结肠组织中NF-κB蛋白表达降低。结论痛泻要方治疗IBS-D肠黏膜低度炎症的作用机制可能与激活结肠FXR表达,抑制结肠组织中NF-κB表达,进而降低血清中促炎因子IL-6、IL-8、TNF-α浓度有关。
Objective To explore the anti-inflammatory mechanism of Tong-Xie-Yao-Fang′s intervention on irritable bowel syndrome with diarrhea(IBS-D),the effects of Tong-Xie-Yao-Fang on farnesol receptor(FXR),nuclear factor-κB(NF-κB)and inflammatory factors in IBS-D model rats were observed.Methods Healthy SD rats were randomly divided into blank group(Control group),model group(Model group),low-dose group of Tong-Xie-Yao-Fang(TXYF-L group,4.977 g/kg),middle-dose group of Tong-Xie-Yao-Fang(TXYF-M group,9.954 g/kg),and high-dose group of Tong-Xie-Yao-Fang(TXYF-H group,19.908 g/kg),Pivium bromide group(PWXA group,0.018 g/kg),with 10 rats in each group.Except for the blank group,the IBS-D rat models of liver depression and spleen deficiency were established by chronic restraint stress and senna intragastric administration.After the model was successfully established,the intervention was continued for 14 days according to the corresponding group treatment method.The general situation,diarrhea rate,fecal moisture content,water injection when abdominal wall retraction reflex reaches 3 scores,sugar water preference rate and serum D-xylose level were observed.The morphological and structural changes of rat colon were observed by HE staining.The expression of FXR gene in colon tissue was detected by real-time fluorescence quantitative PCR,the expression of FXR protein and NF-κB protein in colon tissue was detected by protein Western blot,and the levels of serum IL-6,IL-8 and TNF-αwere detected by ELISA.Results After the treatment of Tong-Xie-Yao-Fang,the weight of each dosage group of Tong-Xie-Yao-Fang increased,the fecal water content decreased,the sugar water preference rate increased,the water injection volume increased when the rats reached the third grade of abdominal retraction reflex score,the serum D-xylose content increased and the food intake increased.HE staining showed that villous or finger-like structures appeared in the mucosal epithelium of model group,and inflammatory cells infiltrated in the upper mucosa.After treatment,the mucosal epithelium in TXYF-M group was smooth,villous or fingerlike structures disappeared,and the infiltration degree of inflammatory cells decreased.Compared with the model group,the expression of FXR mRNA and FXR protein in the colon tissue of rats in the high-dose group of Tong-Xie-Yao-Fang increased significantly,while the concentration of IL-6,IL-8 and TNF-αin serum and the expression of NF-κB protein in the colon tissue of rats in the high-dose group of Tong-Xie-Yao-Fang decreased.Conclusion The mechanism of Tong-Xie-Yao-Fang in treating low-grade inflammation of IBS-D intestinal mucosa may be related to activating the expression of FXR in colon,inhibiting the expression of NF-κB in colon tissue,and then reducing the concentration of pro-inflammatory factors IL-6,IL-8 and TNF-αin serum.
作者
王秋香
李刘英
杨洋
王娟
吴瑞珂
冯培民
WANG Qiuxiang;LI Liuying;YANG Yang;WANG Juan;WU Ruike;FENG Peimin(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Guangyuan Central Hospital,Guangyuan 628000,China;Zigong First People's Hospital,Zigong 643000,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第6期1709-1720,共12页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
四川省中医药管理局面上项目(2023MS230):基于肠道菌群-胆汁酸轴探讨痛泻要方治疗肝郁脾虚型IBS-D模型大鼠的作用机制
负责人:冯培民
四川省中医药管理局面上项目(2024MS606):基于肠道菌群的SCFAs/GPR43/5-HT途径探讨痛泻要方治疗肝郁脾虚型IBS-D的作用机制
负责人:王秋香。
关键词
痛泻要方
腹泻型肠易激综合征
法尼醇受体
调节核因子-κB
炎性因子
Tong-Xie-Yao-Fang
Irritable bowel syndrome with diarrhea
Farnesol receptor
Regulatory nuclear factor-κB
Inflammatory factor
作者简介
通讯作者:冯培民(ORCID:0000-0002-7716-368X),教授,主任医师,博士生导师,主要研究方向:中医药防治自身免疫性疾病及消化系统疾病的临床研究。
