摘要
研究昼夜节律异常对小鼠脾脏免疫功能的影响。采用14月龄的节律基因Per1/Per2双敲除(Per1^(-/-)/Per2^(-/-))雄性C57BL/6小鼠(双敲除组,DKO)与野生型的雄性C57BL/6小鼠(野生组,WT),测量体重、脾脏重量,同时,对脾脏组织进行HE染色、免疫组化、转录组测序以及脾脏B细胞和T细胞的流式细胞检测,分析节律基因敲除所导致的节律异常对小鼠脾脏免疫功能的影响。结果显示:与野生型相比,双敲除组脾脏指数增加,HE染色显示脾脏白髓变薄,动脉周围淋巴鞘消失且边缘区消失;流式细胞检测结果显示,双敲除组的B细胞和T细胞的比例都显著下降。TFR1免疫组化显示DKO小鼠出现铁死亡表现,转录组测序结果显示双敲除组铁死亡途径显著激活,其中参与脂质过氧化的Acsl4、Lpcat3基因表达显著上调,及谷胱甘肽合成的Gclm、Gclc基因表达显著上调。所得结论:Per1/Per2双敲除节律异常小鼠脾脏免疫功能降低,可能涉及铁死亡途径。
To study the effect of abnormal circadian rhythm on spleen immune function of mice.Body weight,spleen weight and tissue morphology of 14-month-old male C57BL/6 mice with Per1/Per2 double knockout(DKO)and wild-type male C57BL/6 mice(wild group)were measured by HE staining,immunohistochemistry and transcriptome sequencing.At the same time,flow cytometry were performed on spleen B cells and T cells.To analyze the effect of abnormal rhythm induced by rhythm gene knockout on spleen immune function of mice.The results showed that compared with the wild type,spleen index was higher in the double knockout group,the spleen white pulp was thinner,the periarterial lymphatic sheath disappeared and the marginal area disappeared.Flow cytometry results showed that the proportions of B cells and T cells in the double knockout group were significantly decreased.TFR1 immunohistochemistry showed ferroptosis in DKO mice spleen.Transcriptome sequencing results showed that Acsl4,Lpcat3,Gclm and Gclc genes were significantly up-regulated,promoting lipid peroxidation and glutathione synthesis.Conclusion:The spleen function of Per1/Per2 double knockout mice with abnormal rhythm decreased,which might be involved in ferroptosis.
作者
张世杰
余佳乐
刘志国
ZHANG Shi-jie;YU Jia-le;LIU Zhi-guo(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China)
出处
《武汉轻工大学学报》
2022年第2期33-37,共5页
Journal of Wuhan Polytechnic University
关键词
节律
铁死亡
免疫力
转录组测序
rhythm
ferroptosis
immunity
transcriptome sequencing
