湖南中医药大学学报2025，Vol.45Issue(11)：2026-2034,9.DOI:10.3969/j.issn.1674-070X.2025.11.002
苦参芩黛汤灌肠对溃疡性结肠炎大鼠Cajal间质细胞自噬的影响
Effects of Kushen Qindai Decoction enema on autophagy of interstitial cells of Cajal in rats with ulcerative colitis
摘要
Abstract
Objective To investigate the effects of Kushen Qindai Decoction(KSQDD)enema on autophagy of colonic interstitial cells of Cajal(ICC)in rats with ulcerative colitis(UC)and its mechanism of action.Methods Forty SPF-grade male SD rats were randomly divided into blank control group(n=10)and experimental group(n=30).The experimental group received enema administration of 5%2,4,6-trinitrobenzene sulfonic acid(TNBS)(100 mg/kg)combined with 50%ethanol to establish an acute UC model.After successful modeling,the experimental group was randomly subdivided into model group,KSQDD group and mesalazine group,with ten rats in each group.The blank control and model groups received enema with an equal volume of normal saline.The KSQDD group received KSQDD enema at a dose of 10.28 g/(kg·d)(crude drug concentration),while the mesalazine group received 0.41 g/(kg·d)enema,once daily for 14 consecutive days.Body weight and disease activity index(DAI)of rats in each group were recorded.Colonic macroscopic damage was observed and colonic mucosal damage index(CMDI)was calculated.Histopathological changes in the colon were assessed by Hematoxylin-eosin(HE)staining.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),IL-1β,and IL-10 were determined by Enzyme-linked immunosorbent assay(ELISA).Immunofluorescence was used to check the co-localization of c-kit and the autophagy-related protein Beclin-1 in colonic tissue.Western blot was used to measure the microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ ratio and the protein expressions of Beclin-1 and p62 in colonic tissue.Transmission electron microscopy(TEM)was used to observe the ultrastructure of ICC and the number of autophagosomes.Results Compared with the blank control group,the model group showed reduced body weight(P＜0.01),elevated DAI and CMDI scores(P＜0.01),significant damage and swelling of the intestinal villi,massive infiltration of neutrophils in the lamina propria and submucosa,obvious edema,and disordered structure of the mucosal layer and submucosa.Serum levels of IL-6,TNF-α,and IL-1β increased(P＜0.01),while IL-10 level decreased(P＜0.01).The co-expression ratio of c-kit and Beclin-1,the LC3-Ⅱ/LC3-Ⅰ ratio,and Beclin-1 protein expression in colonic tissue were up-regulated,while p62 protein expression was down-regulated(P＜0.01).The number of autophagosomes in ICC increased,with obvious organelle damage.Compared with the model group,both the KSQDD and mesalazine groups showed increased body weight(P＜0.01),decreased DAI and CMDI scores(P＜0.05 or P＜0.01),improved intestinal villi damage,reduced neutrophil infiltration in the submucosa and lamina propria,alleviated edema,and improved structural clarity of the mucosal layer and submucosa.Serum levels of IL-6,TNF-α,and IL-1β decreased(P＜0.01),while IL-10 level increased(P＜0.01).The co-expression ratio of c-kit and Beclin-1,the LC3-Ⅱ/LC3-Ⅰ ratio,and Beclin-1 protein expression in colonic tissue were down-regulated(P＜0.01),while p62 protein expression was up-regulated(P＜0.01).The number of autophagosomes in ICC decreased,and organelle damage was alleviated.Compared with the KSQDD group,the mesalazine group demonstrated increased body weight(P＜0.01),decreased DAI and CMDI scores(P＜0.05,P＜0.01),improved intestinal villi damage,reduced neutrophil infiltration in the submucosa and lamina propria,alleviated edema,and improved structural clarity of the mucosal layer and submucosa.Serum levels of IL-6,TNF-α,and IL-1β decreased(P＜0.01),while IL-10 level increased(P＜0.01).The co-expression ratio of c-kit/Beclin-1,the LC3-II/LC3-I ratio,and Beclin-1 protein expression in colonic tissue were down-regulated(P＜0.01),while p62 protein expression was up-regulated(P＜0.01).Organelle damage in ICC was reduced.Conclusion KSQDD enema can effectively alleviate intestinal inflammation in UC rats,and its mechanism may be related to inhibiting excessive autophagy of ICC and regulating the expression of autophagy-related proteins.关键词
溃疡性结肠炎/苦参芩黛汤/灌肠/Cajal间质细胞/自噬Key words
ulcerative colitis/Kushen Qindai Decoction/enema/interstitial cells of Cajal/autophagy分类
医药卫生引用本文复制引用
符佳,张伟,周赛男,徐寅,朱莹..苦参芩黛汤灌肠对溃疡性结肠炎大鼠Cajal间质细胞自噬的影响[J].湖南中医药大学学报,2025,45(11):2026-2034,9.基金项目
国家自然科学基金项目(82374426,81874466) （82374426,81874466）
全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号) （国中医药人教函[2022]75号）
湖南中医药大学中医学国内一流建设学科项目 （）
湖南省中医药科研课题(B2023079) （B2023079）
湖南中医药大学校院联合基金项目(2023XYLH032) （2023XYLH032）
国家中医药管理局中医治疗优势病种(临床循证能力提升)项目(czxm-kyb-2025001). （临床循证能力提升）
