不同形态硒对炎性疼痛大鼠T细胞免疫功能与Foxp3表达的调控及作用机制  

Regulation and mechanism of different forms of selenium on T cell immune function and Foxp3 expression in inflammatory pain rats

作　　者：张天蔚 彭婷 李永寿 李永武 王艳波 ZHANG Tian-Wei;PENG Ting;LI Yong-Shou(Department of Laboratory,the Second Affiliated Hospital of Hainan Medical College,Haikou 570311,Hainan,China)

机构地区：[1]海南医学院第二附属医院检验科,海南海口570311 [2]海南医学院基础医学与生命科学学院,海南海口570311

出　　处：《中国老年学杂志》2023年第11期2698-2704,共7页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81660004);海南省卫生健康行业科研项目(19A200162)。

摘　　要：目的探究L-硒甲基硒代半胱氨酸(L-SeMC)、亚硒酸钠(Na_(2)SeO_(3))对炎性疼痛大鼠免疫功能的影响,并从叉头样蛋白(Fox)p3和核转录因子(NF)-κB途径探索其作用机制。方法采用甲醛诱导大鼠建立炎性疼痛模型,实验分为空白对照组、模型组、L-SeMC组、Na_(2)SeO_(3)组,每组10只。造模24 h后,L-SeMC组灌胃10μg/(kg·d)的L-SeMC,Na_(2)SeO_(3)组灌胃10μg/(kg·d)的Na_(2)SeO_(3),空白对照组和模型组灌胃等体积生理盐水,连续灌胃14 d。检测足跖肿胀率,Von-Frey针刺检测足底疼痛阈值,酶联免疫吸附试验(ELISA)检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6、IL-10水平,分离脾脏单个核细胞并制备悬液,流式细胞术检测T淋巴细胞亚群CD4^(+)、CD8^(+)表达,免疫荧光染色检测脊髓背角Foxp3和离子钙结合衔接分子(Iba)-1表达,Western印迹检测脊髓背角Foxp3和NF-κB信号通路相关蛋白表达水平。结果与空白对照组比较,模型组足跖肿胀率增加,机械痛阈值降低,血清TNF-α、IL-1β、IL-6含量增高,IL-10含量降低,脊髓背角Iba-1、Foxp3蛋白表达上调,p-IκBα/IκBα、p-p65/p65比值均升高,差异有统计学意义(P<0.01)。与模型组比较,L-SeMC组和Na_(2)SeO_(3)组足跖肿胀率下降,机械痛阈值升高,血清TNF-α、IL-1β、IL-6含量降低,IL-10含量升高,脊髓背角Iba-1、Foxp3蛋白表达下调,p-IκBα/IκBα、p-p65/p65比值均下降,差异有统计学意义(P<0.01)。而4组CD4^(+)T细胞、CD8^(+)T细胞比率及CD4^(+)/CD8^(+)比值差异均无统计学意义(P>0.05)。结论L-SeMC和Na_(2)SeO_(3)对大鼠炎性疼痛具有缓解作用,能够减少炎症细胞因子释放,抑制Foxp3表达与NF-κB信号通路激活,但对淋巴细胞亚群CD4^(+)、CD8^(+)表达无影响。Objective To investigate the effects of L-selenomethylselenocysteine(L-SeMC)and sodium selenite(Na_(2)SeO_(3))on inflammatory pain and immune function in rats,and to explore the mechanism of the forkhead box protein(Fox)p3 and nuclear transcription factor(NF)-κB pathways.Methods Formaldehyde-induced inflammatory pain model was established in rats,the experiment was divided into blank control group,model group,L-SeMC group and Na_(2)SeO_(3) group,with 10 rats in each group.24 h after modeling,L-SeMC group was intragaxed with 10μg/(kg·d)L-SeMC,Na_(2)SeO_(3) group was intragaxed with 10μg/(kg·d)Na_(2)SeO_(3),blank control group and model group were intragaxed with equal volume of normal saline for 14 d.Plantar swelling rate was determined,Von-Frey acupuncture was used to detect the threshold of plantar pain,the serum levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6 and IL-10 were determined by enzyme-linked immunosorbent assay(ELISA).The splenic mononuclear cells were isolated and the suspension was prepared,the expressions of CD4^(+)and CD8^(+)in T lymphocyte subsets were detected by flow cytometry.Immunofluorescence staining was used to detect the expressions of Foxp3 and ionized calcium bindingadaptor molecule(IBA)-1 in spinal dorsal horn.Western blot was used to detect the expression levels of Foxp3 and NF-κB signaling pathway-related proteins in spinal dorsal horn.Results Compared with blank control group,the plantar swelling rate was increased,the mechanical pain threshold was decreased,the contents of TNF-α,IL-1βand IL-6 in serum were increased,the content of IL-10 was decreased,the expressions of Iba-1 and Foxp3 protein in spinal dorsal horn were up-regulated,the ratios of p-IκBα/IκBαand p-p65/p65 were increased in model group,the differences were statistically significant(P<0.01).Compared with model group,the plantar swelling rate in L-SeMC group and Na_(2)SeO_(3) group were decreased,the mechanical pain threshold were increased,the content of TNF-α,IL-1βand IL-6 in serum were

关 键 词：炎性疼痛 L-硒甲基硒代半胱氨酸 亚硒酸钠 T细胞免疫 

分 类 号：R402[医药卫生—临床医学]