| 宋立成,闫鹏,杨庆云,等.免疫抑制小鼠脓毒症前后肺部及肠道微生态的变化特点[J].中国临床保健杂志,2021,24(6):820-825. |
| 免疫抑制小鼠脓毒症前后肺部及肠道微生态的变化特点 |
| Changes of lung and intestinal microbiota in immune-suppressed mice before and after sepsis |
| 投稿时间：2021-05-19 |
| DOI：10.3969/J.issn.1672-6790.2021.06.022 |
| 中文关键词: 脓毒症 免疫耐受 微生物学现象 糖皮质激素类 疾病模型,动物 |
| 英文关键词: Sepsis Immune tolerance Microbiological phenomena Glucocorticoids Disease models,animal 〖FL |
| 基金项目:国家重点研发计划项目(2016YFC1304305);中国人民解放军总医院第八医学中心重点项目(2021ZD008) |
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| 中文摘要: |
| 目的 探讨脓毒症前后肠道及肺部微生态的差异。方法 将64只4周龄的雄性C57BL6小鼠用随机数字表法随机分为两组,免疫抑制组(MP-control)小鼠每日腹腔注射甲泼尼龙0.2 mg/g,免疫正常组(control)小鼠每日灌注相同体积的0.9%氯化钠注射溶液,连续灌注30 d,建立免疫抑制模型,记录体质量及检测外周血淋巴细胞亚群变化情况,确定模型建立是否成功。通过气管内及腹腔注射脂多糖(LPS)建立脓毒症模型,记录生存曲线；于建模后第24小时观察外周血炎症因子变化特点,及小鼠肺部及肠道微生态的变化特点。结果 MP-control组小鼠体质量随时间增加呈现为第1周先增高此后逐渐降低的趋势,外周血淋巴细胞亚群表现为总淋巴细胞计数显著降低,CD4/CD8显著降低,CD8细胞在总CD3细胞中的比例显著升高,提示免疫抑制模型建立成功。MP-control组小鼠经LPS干预后,48 h死亡率显著高于control组。肺部微生态分析发现,control组与MP-control组α多样性差异无统计学意义,但MP-control组微生态中抗炎及短链脂肪酸(SCFA)产生相关的菌群明显升高,但MP-control组粪便中微生态出现显著的紊乱；在脓毒症早期,MP-LPS组和LPS-control组肺部微生态α多样性差异无统计学意义,但MP-LPS组促炎相关的菌群水平显著升高,抗炎相关菌群水平显著降低；粪便微生态中,MP-LPS组α多样性显著低于LPS-control组,且抗炎及黏膜屏障相关的菌群水平显著低于LPS-control组,而促炎和机会感染相关菌群水平显著升高。结论 糖皮质激素会对健康小鼠的肺部及肠道微生态造成不同的影响,但当脓毒症出现时,糖皮质激素的长期大量应用会造成更加严重的肺部及肠道微生态紊乱。 |
| 英文摘要: |
| Objective To explore the changes of intestinal and pulmonary microbiota before and after sepsis in immure-suppressed mice.Methods Sixty-four 4-week-old male C57BL6 mice were randomly divided into two groups:MP-control mice were intraperitoneally injected with methylprednisolone 0.2mg/g every day,and control mice were injected with 0.9% sodium chloride injection solution of the same volume every day for 30 days to establish the immunosuppression model.The body weight was recorded and the changes of lymphocyte subsets in peripheral blood were detected to determine whether the model was established successfully.Sepsis model was established by intratracheal and intraperitoneal injection of lipopolysaccharide(LPS),and survival curves were recorded.The changes of inflammatory factors in peripheral blood,lung and intestinal microecology of mice were observed at the 24 h after modeling.Results The body weight of mice in MP-control group increased at the first week and then decreased with the increase of time.The peripheral blood lymphocyte subsets showed a significant decrease in total lymphocyte count,a significant decrease in CD4/CD8,and a significant increase in the proportion of CD8 cells in total CD3 cells,suggesting that the immunosuppression model was established successfully.After LPS intervention,the mortality of MP-LPS group was significantly higher than that of LPS-control group.Pulmonary microecological analysis showed that there was no significant difference inα diversity between the control group and the MP-control group,but the anti-inflammatory and short-chain fatty acid (SCFA) production related microflora in the MP-control group was significantly increased,while the fecal microflora in the MP-control group was significantly disrupted.In the early stage of sepsis,there was no significant difference in pulmonary microbiota α diversity between the MP-LPS group and the LPS-control group,but the level of pro-inflammatory related bacteria was significantly increased and the level of anti-inflammatory related bacteria was significantly decreased in the MP-LPS group.In fecal microbiota,the α diversity of MP-LPS group was significantly lower than that of LPS-control group,and the levels of anti-inflammatory and mucosal barrier related bacteria were significantly lower than that of LPS-control group,while the levels of pro-inflammatory and opportunistic infection related bacteria were significantly higher.Conclusions Glucocorticoids can have different effects on the lung and intestinal microecology of control mice,but when sepsis appears,the long-term use of glucocorticoids can cause more serious pulmonary and intestinal microecology disorders. |
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