基于Nrf2/HO-1和NF-κB信号通路探讨清化止泻方对腹泻型肠易激综合征大鼠巨噬细胞极化的影响  

Effects of Qinghua Zhixie Formula on macrophage polarization in a rat model of diarrhea-predominant irritable bowel syndrome via Nrf2/HO-1 and NF-κB signaling pathways

作　　者：黄华[1] 王永通 丁旭枫 蒋捷[1] 季利江[1] HUANG Hua;WANG Yong-tong;DING Xu-feng;JIANG Jie;JI Li-jiang(Department of Anorectal Surgery,Changshu Hospital Affiliated to Nanjing University of Chinese Medicine,Nanjing 215500,China)

机构地区：[1]南京中医药大学常熟附属医院肛肠科,江苏南京215500

出　　处：《中成药》2025年第2期438-445,共8页Chinese Traditional Patent Medicine

基　　金：江苏省中医药科技发展计划项目(MS2021058,MS2023099);苏州市科技局立项资助(SKY2022072);常熟市科技局立项资助(CS202233);常熟市卫健委立项资助(CSWSQ202304,CSWS202103);镇江市中医脾胃病临床医学研究中心开放课题(SSPW2022-KF08);苏州市卫生健康委员会“科教强卫”面上项目(MSXM2024053);南京中医药大学自然科学基金项目(XZR2020062)。

摘　　要：目的探讨清化止泻方对腹泻型肠易激综合征(IBS-D)大鼠的改善作用及其对M1/M2型巨噬细胞极化的影响。方法SD大鼠随机分为空白组、模型组、匹维溴铵组(20 mg/kg)和清化止泻方低、高剂量组(13.64、27.29 g/kg),每组10只,采用慢性束缚应激加番泻叶水煎剂灌胃法构建IBS-D大鼠模型,各组给予相应剂量药物14 d。观察各组大鼠进食状况、粪便性状和体质量变化;腹部撤离反射(AWR)测定大鼠肠疼痛敏感性;HE染色观察结肠黏膜组织病理学;TUNEL染色观察结肠组织细胞凋亡情况;免疫荧光染色检测大鼠结肠组织ZO-1、Occludin、Nrf2蛋白表达;流式细胞术检测肠系膜淋巴结中M1型和M2巨噬细胞比值;Western blot法检测结肠组织Nrf2、HO-1、p-IκBα、p-P65蛋白表达。结果与模型组比较,清化止泻方各剂量组大鼠体质量、AWR痛觉阈值升高(P<0.05,P<0.01);结肠组织结构基本完整,上皮细胞排列整齐,未见大量细胞变性脱落;结肠组织ZO-1、Occludin蛋白荧光强度增强(P<0.05,P<0.01);结肠组织细胞凋亡率降低(P<0.05,P<0.01);肠系膜淋巴结巨噬细胞向M2极化(P<0.05,P<0.01);结肠组织Nrf2、HO-1蛋白表达升高(P<0.05,P<0.01),p-IκBα/IκBα和p-P65/P65蛋白表达降低(P<0.05,P<0.01)。结论清化止泻方可改善IBS-D大鼠肠上皮黏膜屏障功能,其作用可能与激活Nrf2/HO-1信号通路,抑制NF-κB信号通路活化,调控巨噬细胞向M2极化有关。AIM To investigate the effects of Qinghua Zhixie Formula on improving symptoms of a rat model of diarrhea-predominant irritable bowel syndrome(IBS-D)and its impact on the polarization of M1/M2 macrophages.METHODS The SD rats were randomly divided into the blank group,the model group,the pirenzepine group(20 mg/kg),and the low-dose and high-dose Qinghua Zhixie Formula groups(13.64,27.29 g/kg),with 10 rats in each group.An IBS-D rat model was established using chronic restraint stress combined with gavage of senna leaf decoction.Each group underwent corresponding medication for 14 days.The rats had their changes of food intakes,stool characteristics,and body weight observed;their abdominal withdrawal reflex(AWR)measured to assess the intestinal pain sensitivity;their histopathological changes in the colon mucosa observed using HE staining;their colon mucosa apoptosis assessed using TUNEL staining;their colon expressions of ZO-1,Occludin and Nrf2 proteins detected using immunofluorescence staining;their ratio of M1/M2 macrophages in mesenteric lymph nodes measured by Flow cytometry;and their colon protein expressions of Nrf2,HO-1,p-IκBαand p-P65 analyzed by Western blot.RESULTS Compared with the model group,the groups intervened with Qinghua Zhixie Formula showed increased body weight and AWR pain threshold(P<0.05,P<0.01);generally intact colon tissue structure with orderly arrangement of epithelial cells and no significant cell degeneration or shedding;enhanced fluorescence intensity of colon ZO-1 and Occludin proteins(P<0.05,P<0.01);reduced apoptosis rate of colon tissue cells(P<0.05,P<0.01);more polarized M2 phenotype of macrophages in mesenteric lymph nodes(P<0.05,P<0.01);increased protein expressions of colon Nrf2 and HO-1(P<0.05,P<0.01);and decreased expressions of p-IκBα/IκBαand p-P65/P65(P<0.05,P<0.01).CONCLUSION Qinghua Zhixie Formula can improve the function of the intestinal epithelial mucosal barrier in the IBS-D rat models,and this may be related to the modulation of macrophage polarization toward

关 键 词：清化止泻方 腹泻型肠易激综合征 巨噬细胞 巨噬细胞极化 Nrf2/HO-1信号通路 NF-ΚB信号通路 

分 类 号：R285.5[医药卫生—中药学]