金黄色葡萄球菌核酸酶对2,4,6-三硝基苯磺酸诱导的小鼠结肠炎的保护作用

张婷婷; 梅银柳; 董万法; 王镜勋; 金亮; 吴洁

doi:10.11665/j.issn.1000-5048.20200211

金黄色葡萄球菌核酸酶对2,4,6-三硝基苯磺酸诱导的小鼠结肠炎的保护作用

中国药科大学生命科学与技术学院微基因药学实验室

基金项目: 国家自然科学基金资助项目(No.81973224)；中国药科大学“双一流”创新团队项目资助(No.CPU2018GF/GY16,No.CPU2018GF/GY17)

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出版历程
- 刊出日期: 2020-04-24

Protective effect of staphylococcal nuclease on 2, 4, 6-trinitro-benzene sulfonic acid-induced colitis in mice

摘要

摘要: 探究金黄色葡萄球菌核酸酶(SNase)对2，4，6-三硝基苯磺酸(TNBS)诱导小鼠结肠炎的改善作用及机制。用2.5% TNBS溶液灌肠雌性BALB/c小鼠建立结肠炎模型，并连续6 d灌胃给予以重组乳酸菌为递呈载体的SNase。探究SNase介导的中性粒细胞胞外诱捕网(NETs)的降解对小鼠结肠炎的影响。实验分为正常组、TNBS模型组、NZ9000乳酸菌组、表达SNase的重组乳酸菌组。每日观测小鼠体质量、粪便性状和粪便隐血情况，观察期结束取结肠组织进行HE病理分析，检测结肠组织髓过氧化物酶(MPO)酶活和促炎细胞因子的mRNA表达水平，检测血清炎性细胞因子含量，同时免疫组化检测结肠组织中性粒细胞及其NETs标志物的表达水平。结果表明，乳酸菌递呈的SNase能缓解TNBS诱导的结肠炎小鼠体质量下降，降低疾病活动指数(DAI)评分，缓解结肠缩短并减轻病理损伤，降低结肠组织MPO酶活及炎性细胞因子表达，同时改善了血清炎性水平，免疫组化结果表明结肠组织Ly6G和citH3水平下降。初步机制表明，SNase能够下调炎性细胞因子的表达，降低NETs水平从而缓解小鼠结肠炎。
- 金黄色葡萄球菌核酸酶 /
- 乳酸乳球菌 /
- 2,4,6-三硝基苯磺酸 /
- 中性粒细胞胞外诱捕网 /
- 结肠炎
Abstract: To explore the improving effect and mechanism of staphylococcal nuclease(SNase)-mediated degradation of neutrophil extracellular traps(NETs)on 2, 4, 6-trinitro-benzene sulfonic acid(TNBS)-induced colitis in mice. The model of colitis in female BALB/c mice was established by intrarectal injection of 2. 5% TNBS solution, and SNase loaded by Lactococcus lactis(L. lactis)were orally administrated for 6 days. To investigate the effect of SNase-mediated degradation of neutrophil extracellular traps on colitis in mice, the experiment was divided into control group, TNBS model group, NZ900 group and L. lactis pCYT: SNase group. The daily body weight, stool consistency and bleeding of mice were observed. The pathological condition of HE in colon group was detected. The activity of MPO and the mRNA expression level of inflammatory cytokines in each group were measured, and the concentration of inflammatory factors in serum was detected. The expression of NETs level marker citH3 in colon tissue was determined by immunohistochemistry. The results showed that SNase loaded by lactis acid bacteria could alleviate the weight loss, disease activity index score, colonic length and pathological damage induced by TNBS in mice, and reduce the levels of inflammatory cytokines in serum and colonic tissue, inhibit the activity of MPO and the expression of Ly6G and citH3 in colon tissue. The preliminary mechanism showed that SNase could down-regulate the expression of inflammatory cytokines and reduce the content of NETs markers to alleviate colitis in mice.
- staphylococcal nuclease /
- Lactococcus lactis /
- 2,4,6-trinitro-benzene sulfonic acid /
- neutrophil extracellular traps /
- colitis

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