环磷酰胺诱导的小鼠免疫低下模型的方法学考察及金樱子提取物联合益生菌调节免疫功能的研究
Methodological investigation of cyclophosphamide-induced immunocompro-mised mice model and effect of extract of Rosa laevigata Michx.combined with probiotics on immune function
刘泽坤 1马俊丽 1李艳 1郑宁宁 1盛丽莉 1李后开1
作者信息
- 1. 上海中医药大学中药学院,上海 201203
- 折叠
摘要
目的:建立更符合人体免疫低下状态的小鼠模型,并基于此模型考察金樱子、益生菌以及二者联合用药对免疫功能的调节作用.方法:采用不同给药剂量、时间和实验终点的环磷酰胺建立小鼠免疫低下模型,并与氢化可的松及环孢素A诱导的免疫低下模型对比,确立了25 mg/kg环磷酰胺隔日共注射7次,无恢复期的免疫低下小鼠模型;流式细胞术检测小鼠脾脏淋巴细胞(CD4+T细胞、CD8+T细胞、CD19+B细胞)比例变化;qRT-PCR检测小鼠脾脏炎症因子(IL-1β、IL-6)mRNA表达.结果:80 mg/kg环磷酰胺连续注射3 d后,胸腺萎缩,脾脏肿大,脾脏中CD4+T细胞、CD8+T细胞、CD19+B细胞比例下降,IL-1β及IL-6 mRNA表达上调;25 mg/kg环磷酰胺隔日注射共7次后,胸腺和脾脏萎缩,脾脏中CD4+T及CD8+T细胞比例呈升高趋势,IL-1β及IL-6 mRNA表达上调;环磷酰胺造模后恢复1周或2周,小鼠免疫器官重量和指数、脾脏淋巴细胞比例、脾脏炎症细胞因子mRNA表达均显著恢复;金樱子联合益生菌可显著恢复环磷酰胺导致的CD4+T细胞上调以及炎症因子IL-1β上调.结论:本研究建立了25 mg/kg环磷酰胺隔日注射共7次,无恢复期的免疫低下小鼠模型,并基于此模型发现金樱子联合益生菌具有一定的正向免疫调节作用.
Abstract
Objective:To establish a mice model that is more in line with the human immunocompromised state,and to inves-tigate the regulatory effect of Rosa laevigata Michx.,probiotics and their combination on their immune function.Methods:Immunosup-pressive model of mice was established by using cyclophosphamide with different dosage,time and end point,and compared with im-munosuppressive model induced by hydrocortisone and cyclosporine A,immunosuppressive model of mice was established by injecting 25 mg/kg cyclophosphamide 7 times every other day,without recovery period;changes in the proportion of lymphocytes in spleen(CD4+T cells,CD8+T cells,CD19+B cells)were analyzed by flow cytometry;qRT-PCR was used to detect expression levels of inflam-matory factors(IL-1β and IL-6)mRNA in spleen.Results:After continuous injection of 80 mg/kg cyclophosphamide for 3 days,the thymus was atrophied,the spleen was significantly swelling,proportion of immune cells CD4+T cells,CD8+T cells and CD19+B cells in spleen were significantly decreased,while mRNA expressions of IL-1β and IL-6 were up-regulated.After 7 injections of 25 mg/kg cyclophosphamide every other day,the thymus and the spleen were significantly atrophied,proportion of CD4+T cells and CD8+T cells in spleen showed an increasing trend,and mRNA expressions of IL-1β and IL-6 were up-regulated;after 1 week or 2 weeks recovery period from cyclophosphamide injection,weight and index of immune organs,proportion of spleen lymphocytes and mRNA expres-sions of inflammatory cytokines in spleen were significantly reversed;Rosa laevigata Michx.combined with probiotics could signifi-cantly reverse the up-regulation of CD4+T cells and inflammatory factor IL-1β caused by cyclophosphamide.Conclusion:In this study,an immunocompromised mice model has been established with 7 injections of 25 mg/kg cyclophosphamide every other day and without recovery period.And a certain positive immunomodulatory effect of the combination of Rosa laevigata Michx.and probiotics has been found.
关键词
免疫功能低下/环磷酰胺/动物模型/药物评价Key words
Immunocompromised/Cyclophosphamide/Animal models/Drug evaluation引用本文复制引用
基金项目
上海市优秀学术带头人计划项目(21XD1403500)
出版年
2024
国家科技期刊平台
NSTL
万方数据