间充质干细胞对自然衰老脓毒症大鼠炎性因子及T淋巴细胞亚群的影响  

Effect of mesenchymal stem cells on inflammatory factors and T-lymphocyte subsets in naturally aged sepsis rats

作　　者：王陆 邓子辉[3] 赵妍[2] 李云 刘育妍 杨萌萌[2] 周飞虎[2] 康红军[2] Wang Lu;Deng Zi-Hui;Zhao Yan;Li Yun;Liu Yu-Yan;Yang Meng-Meng;Zhou Fei-Hu;Kang Hong-Jun(Medical School of Chinese PLA,Beijing 100853,China;Department of Critical Care Medicine,the First Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Department of Basic Medicine,Graduate School,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院,北京100853 [2]解放军总医院第一医学中心重症医学科,北京100853 [3]解放军总医院研究生院基础医学教研室,北京100853

出　　处：《解放军医学杂志》2023年第5期545-551,共7页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81671966)。

摘　　要：目的 探讨间充质干细胞(MSCs)对自然衰老脓毒症大鼠炎性因子及T淋巴细胞亚群的影响。方法 将90只自然衰老的SD大鼠(21月龄)按随机数字表法分为3组,其中模型组36只,治疗组36只,假手术组18只,各组大鼠再随机平均分为3组,分别在造模或治疗后6、24和72 h麻醉断头处死取材。采用盲肠结扎穿孔法(CLP)建立脓毒症模型。假手术组除不行盲肠结扎穿孔法外,其他操作同模型组。治疗组大鼠在造模1 h后,经尾静脉注射脐带MSCs,模型组和假手术组在造模1 h后经尾静脉注射等量的生理盐水。观察各组大鼠72 h生存率,通过Aimplex法检测白细胞介素(IL)-1β、IL-6、IL-10、IL-17A、肿瘤坏死因子(TNF)-α及转化生长因子(TGF)-β的水平。对各组大鼠脾组织行TUNEL染色,激光共聚焦显微镜观察脾淋巴细胞凋亡情况;流式细胞术分析各组大鼠CD4^(+)/CD8^(+)T细胞比值。结果 假手术组大鼠观察期内无异常,术后72 h存活率为100.00%。模型组大鼠术后出现竖毛、食欲不振及精神萎靡等症状,而这些症状在治疗组大鼠中有所缓解,且治疗组CLP术后72 h存活率明显高于模型组(66.67%vs. 41.67%,P<0.01)。与假手术组相比,模型组IL-1β、IL-6和IL-17A等促炎因子水平升高,脾结构模糊、淋巴细胞凋亡比例增加,外周血和脾中CD4^(+)/CD8^(+)T淋巴细胞比值降低,差异均有统计学意义(P<0.05);与模型组相比,治疗组IL-1β、IL-6和IL-17A等促炎因子水平降低,抗炎因子IL-10水平升高,脾结构清晰、淋巴细胞凋亡明显减少,外周血和脾中CD4^(+)/CD8^(+)T淋巴细胞比值升高,差异均有统计学意义(P<0.05)。结论 MSCs能够明显提高自然衰老脓毒症大鼠的生存率,抑制过度的炎症反应,减少淋巴细胞凋亡,改善机体免疫功能,从而发挥保护作用。Objective To investigate the effects of mesenchymal stem cells(MSCs)on inflammatory factors and T-lymphocyte subsets in naturally aged sepsis rats.Methods Ninety naturally aged SD rats(21 months old)were divided into 3 groups according to the random number table method:36 rats in the model group,36 rats in the treatment group,and 18 rats in the sham group.In each group,animals were then randomly divided equally into three sub-groups.The rats were severed and executed after 6-h,24-h,and 72-h anesthesia for modeling or treatment,respectively.The sepsis model was established by the cecum ligation perforation(CLP)method.The sham group was operated as the model group,except that the cecum ligation and perforation method was not used.After 1-hour of modeling,the rats in the treatment group were injected with umbilical cord MSCs through the tail vein,and the rats in the model group and the sham group were injected with the same volume of saline through the tail vein.The 72-hour survival rate of rats in each group was observed,and the levels of interleukin(IL)-1β,IL-6,IL-10,IL-17A,tumor necrosis factor(TNF)-α,and transforming growth factor(TGF)-βwere measured by Aimplex method.The spleens of rats in each group were stained with TUNEL and the apoptosis of spleen lymphocytes was observed by laser confocal microscopy;the ratio of CD4^(+)/CD8^(+)T cells in each group was analyzed by flow cytometry.Results The rats in the sham group showed no abnormalities during the observation period,and the survival rate at 72-hour after surgery was 100.00%.The rats in the model group showed symptoms such as vertical hair,loss of appetite,and depression after surgery,while these symptoms were improved in the treated rats,and the survival rate at 72-hour after CLP surgery was significantly higher in the treatment group than in the model group(66.67%vs.41.67%,P<0.01).Compared with the sham group,the model group showed increased levels of IL-1β,IL-6,and IL-17A,blurred spleen structure,increased proportion of apoptotic lymphocytes,and decr

关 键 词：脓毒症 间充质干细胞 T淋巴细胞亚群 炎症 自然衰老大鼠 

分 类 号：R631[医药卫生—外科学]