摘要
目的探讨塞来昔布对实验性自身免疫性脑脊髓炎(EAE)模型小鼠脊髓中Th1/Th17细胞分化的抑制作用。方法将36只C57BL/6小鼠随机分为对照组(等体积生理盐水)、模型组(等体积生理盐水)、塞来昔布组(5 mg/kg),各12只。皮下注射髓磷脂少突胶质细胞糖蛋白35-55(MOG_(35-55))混合乳剂以复制EAE小鼠模型。建模成功后,各组小鼠灌胃给予相应药物或生理盐水,每天1次,连续35 d。对小鼠进行神经功能缺损评分;光学显微镜下观察小鼠脊髓髓鞘脱失程度;采用流式细胞仪测定小鼠脊髓中CD_(4)^(+)T细胞与CD_(8)^(+)T细胞占比,分析CD_(4)^(+)T细胞的分化程度;采用酶联免疫吸附法测定小鼠脊髓中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)表达水平;采用Western blot法测定小鼠脊髓中环氧合酶2(COX-2)、核因子κB(NF-κB)、含NLR家族Pyrin域蛋白3(NLRP3)蛋白表达水平。结果与模型组比较,塞来昔布组小鼠同时间点神经功能缺损评分显著降低,小鼠脊髓中CD_(4)^(+)T细胞占比,CD_(4)^(+)T细胞中Th1与Th17细胞占比,TNF-α,IL-6,IL-1β水平,以及COX-2,p-NF-κB,NLRP3蛋白表达水平均显著降低(P<0.05)。模型组小鼠脊髓髓鞘脱失严重,塞来昔布组虽可见部分髓鞘破坏,但整体染色更深,且髓鞘空泡区域较少。结论塞来昔布可在一定程度上改善模型小鼠EAE症状,其机制可能与抑制COX-2活性,减少CD_(4)^(+)T细胞向Th1与Th17亚型分化,降低TNF-α,IL-6,IL-1β等炎性细胞因子水平有关。
Objective To investigate the inhibition effect of celecoxib on Th1/Th17 Cells differentiation in spinal cord of experimental autoimmune encephalomyelitiss(EAE)model mice.Methods A total of 36 C57BL/6 mice were randomly divided into the control group(equal volume of normal saline),the model group(equal volume of normal saline),the celecoxib group(5 mg/kg),with 12 mice in each group.Myelin oligodendrocyte glycoprotein 35-55(MOG_(35-55))mixed emulsion was subcutaneously injected to replicate the mouse model of EAE.After successful modeling,mice in each group were gavaged with corresponding drugs or normal saline once a day for 35 d.Neurological deficit of mice was scored,demyelination degree in spinal cord of mice was observed by optical microscope.Proportion of CD_(4)^(+)T cells and CD_(8)^(+)T cells in spinal cord of mice were measured and the differentiation degree of CD_(4)^(+)T cells was analyzed by flow cytometry.The expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)in spinal cord of mice were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of cyclooxygenase-2(COX-2),nuclear factor-κB(NF-κB),NLR family pyrin domain containing 3(NLRP3)protein in spinal cord of mice were detected by Western blot.Results Compared with those in the model group,the neurological deficit score of mice in the same time point was decreased significantly in the celecoxib group.Proportion of CD_(4)^(+)T cells,proportion of Th1 and Th17 cells in CD_(4)^(+)T cells,the expression levels of TNF-α,IL-6 and IL-1β,the expression levels of COX-2,p-NF-κB,NLRP3 protein decreased significantly in spinal cord of mice(P<0.05).Demyelination of spinal cord of mice in the model group was severe,while partial myelin destruction was observed in the celecoxib group,the overall staining was deeper,and there were fewer vacuoles area in myelin sheath.Conclusion Celecoxib can improve the symptom of EAE in model mice to some extent,the mechanism of which could be related to the inhibition of COX-2 activity,reduction of the differentiation from CD_(4)^(+)T cells to Th1 and Th17 subtypes and the levels of inflammatory cytokines such as TNF-α,IL-6,IL-1β.
作者
朱燕
谢晓辉
焦敏
吴建华
ZHU Yan;XIE Xiaohui;JIAO Min;WU Jianhua(People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang,China 830001;Xinjiang Uygur Autonomous Region Institute of Clinical Pharmacy,Urumqi,Xinjiang,China 830001)
出处
《中国药业》
CAS
2022年第8期47-52,共6页
China Pharmaceuticals
基金
新疆维吾尔自治区自然科学基金[2020D01C104]。
