酪酸梭菌通过调控内源性硫化氢酶影响小鼠DSS结肠炎肠道屏障  

Clostridium butyricum affects intestinal barrier of mice with DSS-induced colitis via regulation of endogenous hydrogen sulfide enzymes

作　　者：沈媛媛 程凯 李丽霞 孙华波 SHEN Yuanyuan;CHENG Kai;LI Lixia;SUN Huabo(Department of Gastroenterology and Hepatology,Yantai Affiliated Hospital of Binzhou Medical University,Yantai 264100,China;Department of Thyroid and Breast Surgery,Yantai Affiliated Hospital of Binzhou Medical University,Yantai 264100,China;Department of Pathology,Yantai Affiliated Hospital of Binzhou Medical University,Yantai 264100,China)

机构地区：[1]滨州医学院烟台附属医院消化内科,山东烟台264100 [2]滨州医学院烟台附属医院甲状腺乳腺外科,山东烟台264100 [3]滨州医学院烟台附属医院病理科,山东烟台264100

出　　处：《中国病理生理杂志》2025年第1期90-96,共7页Chinese Journal of Pathophysiology

基　　金：山东省中医药科技项目(No.M-2023262)。

摘　　要：目的:探讨酪酸梭菌(Clostridium butyricum)调控内源性硫化氢酶对葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的小鼠结肠炎肠道屏障的影响及作用机制。方法:将40只雄性C57BL/6J小鼠随机分为对照组、DSS结肠炎组、低剂量(1×10^(10)CFU/L)酪酸梭菌组和高剂量(1×10^(11)CFU/L)酪酸梭菌组,每组10只。小鼠自由饮用含3.5%DSS的饮用水建立DSS结肠炎模型。酪酸梭菌组小鼠给予10 mL/kg酪酸梭菌溶液灌胃,每日1次,连续14 d。实验结束后,记录各组小鼠疾病活动度指数(disease activity index,DAI)和结肠长度;HE染色观察结肠组织病理形态;qRT-PCR检测结肠组织中内源性硫化氢酶[胱硫醚β-合成酶(cystathionineβ-synthase,CSE)和胱硫醚γ-裂解酶(cystathionineγ-lyase,CBS)]及黏液屏障标志物Lypd8(LY6/PLAUR domain containing 8)的mRNA表达;Western blot检测结肠组织CSE、CBS及巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)蛋白表达;免疫组化法检测结肠组织紧密连接蛋白[闭锁小带蛋白1(zonula occludens-1,ZO-1)和闭合蛋白(occludin)]表达。结果:与对照组比较,DSS结肠炎组小鼠DAI显著升高,结肠长度显著缩短(P<0.05);肠黏膜上皮缺失,隐窝排列紊乱,大量炎症细胞浸润。与DSS结肠炎组相比,酪酸梭菌组小鼠DAI及结肠缩短程度显著下降(P<0.05);肠黏膜上皮相对完整,炎症细胞浸润减少,隐窝排列紊乱减轻;结肠组织中CSE和CBS的mRNA表达水平显著下降,Lypd8的mRNA表达水平显著升高(P<0.05);Western blot检测CSE、CBS和MIF蛋白表达下调;免疫组化检测ZO-1和occludin表达增多。结论:酪酸梭菌能够显著抑制小鼠DSS结肠炎,主要通过下调内源性硫化氢酶CSE和CBS表达,改善肠黏液屏障,维持肠上皮完整性。AIM:To investigate the effect of Clostridium butyricum on the intestinal barrier of mice with dextran sodium sulfate(DSS)-induced colitis,and to explore its mechanism related to the regulation of endogenous hydrogen sulfide enzymes.METHODS:Forty male C57BL/6J mice(ten per group)were randomly divided into control group,DSS-induced colitis group,and two groups with different doses(1×10^(10)CFU/L and 1×10^(11)CFU/L)of Clostridium butyri⁃cum.To establish the colitis model,mice were given distilled water containing 3.5%DSS.The mice in Clostridium butyri⁃cum groups received a 10 mL/kg Clostridium butyricum solution by gastric intubation once per day for 14 d.At the end of the experiment,the disease activity index(DAI)and colon length of the mice from each group were recorded.The colon tissue morphology was determined using HE staining,and qRT-PCR was used to measure the mRNA expression levels of two endogenous hydrogen sulfide enzymes,cystathionineβ-synthase(CBS)and cystathionineγ-lyase(CSE),and the mucus marker LY6/PLAUR domain containing 8(Lypd8).The protein levels of CSE,CBS and macrophage migration inhibitory factor(MIF)in the colon were measured by Western blot.Finally,the expression of tight junction proteins,zonula occludens-1(ZO-1)and occludin,in the colon were evaluated by immunohistochemical staining.RESULTS:Compared with control group,the mice in DSS-induced colitis group had a significant increase in DAI and a decrease in colon length(P<0.05).Furthermore,the DSS-induced colitis mice exhibited loss of colonic mucosal epithelium,disordered crypt arrangement,and extensive infiltration of inflammatory cells.Compared with DSS-induced colitis group,both DAI and colon shortening were significantly attenuated in the groups with different doses of Clostridium butyricum(P<0.05).The mice in Clostridium butyricum groups also exhibited relatively intact colonic mucosal epithelial structure with reduced inflammatory cell infiltration and reversed crypt disorder.The mRNA expression of CSE and CBS was significantly

关 键 词：酪酸梭菌 内源性硫化氢酶 结肠炎 肠道屏障 

分 类 号：R574.62[医药卫生—消化系统] Q936[医药卫生—内科学] R363[医药卫生—临床医学]