免疫抑制继发艰难梭菌相关性腹泻C57BL/6小鼠模型的建立方法研究  

Establishment of C57BL/6 Mouse Model of Clostridium Difficile-Associated Diarrhea Secondary to Immunosuppression

作　　者：马富智 李萌 于小红[3] 朱琳 阿依曼·叶尔江 赵奎君[3] 章从恩[3] 马致洁 MA Fuzhi;LI Meng;YU Xiaohong;ZHU Lin;AYIMAN Yeerjiang;ZHAO Kuijun;ZHANG Congen;MA Zhijie(School of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China;Dept.of Pharmacy,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Dept.of Pharmacy,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]云南中医药大学中药学院,昆明650500 [2]首都医科大学附属北京地坛医院药学部,北京100015 [3]首都医科大学附属北京友谊医院药学部,北京100050

出　　处：《中国医院用药评价与分析》2025年第1期6-13,共8页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：国家重点研发计划资助(No.2023YFC2308200);国家自然科学基金资助项目(No.82374053);高层次公共卫生人才建设项目(No.学科带头人-02-28)。

摘　　要：目的:建立免疫抑制继发艰难梭菌相关性腹泻(CDAD)C57BL/6小鼠模型,为今后新药研究提供更有针对性的动物模型选择。方法:采用C57BL/6小鼠,分为空白组、免疫抑制CDAD模型组(DXMS+MOD组)、免疫正常CDAD模型组(MOD组)、地塞米松组和混合抗菌药物组。在造模后第1、8日2个时间点取材,通过脾脏和胸腺指数、血常规检测、免疫细胞占比、细胞因子分泌水平以及分泌型免疫球蛋白A(sIgA)含量分析各组小鼠免疫状态情况;通过体重变化率、死亡率、临床评分、粪便黏稠度评分、毒素水平检测和结肠病理变化评价模型组小鼠的CDAD病情。结果:与MOD组比较,DXMS+MOD组小鼠造模后第1、8日的脾脏指数(P<0.001,P<0.05)、胸腺指数(P<0.001)显著降低;白细胞计数(P<0.01)、淋巴细胞百分比(P<0.001)显著降低,中性粒细胞百分比(P<0.001)显著升高;B细胞(P<0.001)、CD4^(+)T细胞(P<0.001,P<0.01)比例和CD4^(+)/CD8^(+)(P<0.001)显著降低,CD8^(+)T细胞比例(P<0.001)显著升高,上述差异均有统计学意义。与MOD组比较,DXMS+MOD组小鼠造模后白细胞介素6(第1日:P<0.001;第8日:P<0.01)、单核细胞趋化蛋白1(第1日:P<0.001)、肿瘤坏死因子α(第8日:P<0.01)和γ干扰素(第8日:P<0.01)水平显著降低,差异均有统计学意义。造模后第1、8日,DXMS+MOD组小鼠sIgA含量较MOD组呈现降低趋势,但差异无统计学意义(P>0.05)。相比于MOD组,DXMS+MOD组小鼠体重降低更明显,且两组在造模后第2日(P<0.01)、第3日(P<0.05)和第4日(P<0.05)的差异有统计学意义。造模后,DXMS+MOD组小鼠的粪便黏稠度评分(第5日:P<0.05)、临床症状评分(第1日:P<0.05;第3、5和7日:P<0.001)始终高于MOD组,差异均有统计学意义。造模后第1、8日,DXMS+MOD组的艰难梭菌负载量较MOD组显著升高,差异有统计学意义(P<0.01)。结肠病理结果显示,DXMS+MOD组肠道屏障破坏更严重且出现死亡情况。结论:DXMS+MOD组小鼠可在整个造模过程�OBJECTIVE:To establish the C57BL/6 mouse model of Clostridium difficile-associated diarrhea(CDAD)secondary to immunosuppression,so as to provide a more targeted selection of animal models for future new drug research.METHODS:C57BL/6 mice were extracted to be divided into the blank group,immunosuppressed CDAD model group(DXMS+MOD group),immune-normal CDAD model group(MOD group),dexamethasone group and mixed antibiotics group.Samples were taken at the 1st and 8th days after modelling,the immune status of mice in each group were analyzed by spleen and thymus indexes,routine blood tests,immune cell percentage,cytokine secretion level and secretory immunoglobulin A(sIgA)content.The CDAD condition of model mice were evaluated through the change rates of body weight,mortality rates,clinical scores,faecal consistency scores,detection of toxin levels and pathological changes of colon.RESULTS:Compared with the MOD group,the DXMS+MOD group had significantly lower spleen(P<0.001,P<0.05)and thymus indexes(P<0.001),significantly lower white blood cell counts(P<0.01)and lymphocyte percentages(P<0.001),significantly higher neutrophilic granulocytes percentages(P<0.001);significantly lower B cells ratios(P<0.001),CD4^(+)T cells ratios(P<0.001,P<0.01)and CD4^(+)/CD8^(+)(P<0.001),and significantly higher CD8^(+)T cells ratios(P<0.001)at the 1st and 8th days after modelling,the differences were statistically significant.Compared with the MOD group,the IL-6(P<0.001 at the 1st day,P<0.01 at the 8th day),MCP-1(P<0.001 at the 1st day),TNF-α(P<0.01 at the 8th day)and IFN-γ(P<0.01 at the 8th day)levels in the DXMS+MOD group were significantly lower,with statistically significant differences.At the 1st and 8th days after modelling,the sIgA content in the DXMS+MOD group showed a decreasing compared with the MOD group,while the difference was not statistically significant(P>0.05).Compared with the MOD group,the mice in the DXMS+MOD group showed a more obvious decrease in body weight,and the differences between two groups were statistically

关 键 词：艰难梭菌相关性腹泻 免疫低下状态 动物模型 C57BL/6小鼠 

分 类 号：R975[医药卫生—药品] R978.1[医药卫生—药学]