针刺研究2025，Vol.50Issue(3)：277-286,10.DOI:10.13702/j.1000-0607.20240402
电针调控NLRP3/Caspase-1/GSDMD信号通路抑制细胞焦亡改善溃疡性结肠炎小鼠结肠黏膜屏障损伤
Electroacupuncture improves colonic mucosal barrier damage by regulating NLRP3/Caspase-1/GSDMD signaling pathway and inhibiting pyroptosis in ulcerative colitis mice
摘要
Abstract
Objective To observe the effect of electroacupuncture(EA)at"Tianshu"(ST25)and"Zusanli"(ST36)on NOD-like receptor protein 3(NLRP3)/cysteine aspartate specific protease 1(Caspase-1)/gasdermin D(GSDMD)pyroptosis signaling pathway in ulcerative colitis(UC)mice,so as to explore its mechanism in improving UC by protecting intestinal barrier.Methods Fifty male C57BL/6 mice were randomly divided into normal,model,EA,sham acupuncture and medication groups,with 10 mice in each group.The UC mouse model was established by 3%DSS solution free drinking for 7 consecutive days.The mice in the EA group received EA(2 Hz/10 Hz,0.2 mA)at bilateral ST25 and ST36 for 20 min,while the mice in the sham acupuncture group received only sham acupuncture(light and shallow acupunture at ST25 and ST36).Mice in the medication group were orally administered mesalazine(33.4 g/kg)solution.All the interventions were performed once daily for a total of 7 days.The changes of body weight,stool shape and hematochezia of mice,the disease activity index(DAI)score,and the length of colon were recorded.The intestinal mucosal permeability was observed by in vivo small animal imaging system.HE staining was used to observe the pathological changes of colon tissue.TUNEL staining was used to observe the apoptosis of colon cells.The contents of interleukin(IL)-18 and IL-1β in serum were detected by ELISA.The average fluorescence intensity of tumor necrosis factor-α(TNF-α)and IL-6 in colon tissue was detected by immunofluorescence.The positive expression of IL-18 and IL-1β in colon tissue was detected by immunohistochemistry.The relative expression levels of zonula occludens-1(ZO-1),Occludin,NLRP3,Caspase-1 and GSDMD in colon tissue were detected by Western blot.The mRNA expressions of NLRP3,Caspase-1 and GSDMD in colon tissue were detected by qPCR.Results After modeling,the DAI,inflammatory infiltration,number of apoptotic cells,fluorescence intensity of TNF-α,IL-6 and FITC,positive expression rate of IL-18 and IL-1β,mRNA and protein expression levels of NLRP3,Caspase-1 and GSDMD,serum IL-18 and IL-1β contents were significantly increased(P<0.01)in the model group relevant to the normal group.At the same time,the colon length,ZO-1 and Occludin protein expression were significantly reduced(P<0.01).The increased DAI,inflammatory infiltration,number of apoptotic cells,fluorescence intensity of TNF-α,IL-6 and FITC,positive rate of IL-18 and IL-1β,mRNA and protein expression levels of NLRP3,Caspase-1 and GSDMD,serum IL-18 and IL-1β contents,and the decreased colon length,ZO-1 and Occludin protein expression were all reversed after the EA and medication interventions compared with the model group and in the EA group than in the sham acupuncture group(P<0.01,P<0.05).Conclusion EA can protect the intestinal mucosal permeability in UC mice,which may be related to its functions in regulating the NLRP3/Caspase-1/GSDMD axis and inhibiting pyroptosis,thereby alleviating the inflammatory injury of intestinal mucosa.关键词
电针/溃疡性结肠炎/细胞焦亡/肠道屏障Key words
Electroacupuncture/Ulcerative colitis/Pyroptosis/Intestinal barrier引用本文复制引用
王熙涵,王天朗,王希宁,刘继东,曲怡,董佳梓,王建波,薛亚楠,李阳,张家豪,金璐..电针调控NLRP3/Caspase-1/GSDMD信号通路抑制细胞焦亡改善溃疡性结肠炎小鼠结肠黏膜屏障损伤[J].针刺研究,2025,50(3):277-286,10.基金项目
国家自然科学基金项目(No.81603704) （No.81603704）
辽宁省教育厅高等学校基本科研项目(No.LJKZ0886) （No.LJKZ0886）
辽宁省科学技术计划项目(No.2021-MS-248) （No.2021-MS-248）
