OK-432联合聚肌胞诱导小鼠树突状细胞成熟及其抗损伤作用  

Effect of OK-432 and PolyI:C on maturation of mouse Dendritic Cells and Against Injury

作　　者：王静[1] 倪秀雄[2] 林岷 姚琦[2] 曾真[2] 陈炜[2] WANG Jing;NI Xiu-xiong;LIN Min;YAO Qi;ZENG Zhen;CHEN Wei(Department of Pathology,Xiamen Hospital of Traditional Chinese Medicine,Xiamen 361009,China;Department of Physiology and Pathophysiology,Fujian Medical University,Fuzhou 350004,China)

机构地区：[1]厦门市中医院病理科,福建厦门361009 [2]福建医科大学生理学与病理生理学系,福建福州350004

出　　处：《海峡药学》2021年第3期25-29,共5页Strait Pharmaceutical Journal

基　　金：福建省自然科学基金项目(No.C0410020)。

摘　　要：目的比较体外单独或联合应用TLR配体-沙培林(OK-432)、聚肌胞对树突状细胞(DCs)成熟状态的影响,并将DCs与小鼠前胃癌细胞上清液共培养,检测DCs存活率,探讨OK-432联合聚肌胞对小鼠前胃癌细胞上清液诱导树突状细胞损伤的保护作用及机制。方法采用重组小鼠粒-巨噬细胞集落刺激因子(rmGM-CFS)联合重组小鼠白细胞介素-4(rmIL-4),诱导小鼠骨髓源性的单个核细胞分化为未成熟的DCs(imDCs),分别经OK-432、聚肌胞或OK-432联合聚肌胞诱导DCs成熟,未成熟或成熟的DCs分别与不同浓度的小鼠前胃癌细胞上清液共培养24 h。电镜及光镜观察DCs的细胞形态变化,流式细胞仪检测DCs CD83和CD86的表达情况,ELISA法测定上清液中IL-12的分泌水平,MTT法测定DCs体外刺激同种混合淋巴细胞增殖的免疫活性以及DCs与不同浓度小鼠前胃癌细胞上清液共培养24 h后的活性,免疫组化法检测DCs锌指蛋白A20(A20)的表达水平。结果联合刺激组DCs的形态学,表面抗原CD83和CD86的表达率,IL-12的分泌量及刺激淋巴细胞增殖的能力均显著高于其它处理组;小鼠前胃癌细胞上清液能损伤DCs,联合刺激组DCs的成活率均高于其它处理组,且其成活率与A20蛋白的表达量成正比。结论OK-432联合聚肌胞能有效诱导DCs成熟;OK-432联合聚肌胞对小鼠前胃癌细胞上清液诱导树突状细胞的损伤有保护作用,其机制与上调A20蛋白表达增高有关。OBJECTIVE To investigate the effect of OK-432 and Poly(I:C)on the maturation of murine bone marrow-derived dendritic cells and co-culture with supernatants of Mouse forestomach carcinoma(MFC).To study the protective effect of OK-432 in a combination with PolyI:C on DCs against injury and to speculate the possible mechanism of this function.METHODS The immature DCs were generated from mouse bone marrow monocytes in the presence of recombinant mouse GM-CSF and recombinant mouse IL-4.These DCs were then pulsed separately with OK-432 and Poly(I:C)or in combination.DCs were co-cultured with different concentrations of MFC supernatants for 24 hours.Morphological characters of DCs were observed under the electron microscope.DCs surface molecules CD83 and CD86 were assayed by flow cytometry.The IL-12 levels in supernatant were assayed by ELISA.The appearance of dendritic cells after co-cultured with MFC supernatants for 24 hours under optical microscope.Antigen presenting ability of DCs in allo-MLR and survival rate of DCs were measured by MTT assay.Zinc finger protein A20(A20)expression in DCs was determined by immunohistochemical procedures.RESULTS The DCs pulsed with OK-432 and Poly(I:C)showed typically morphological characters,expressed much more antigens CD83 and CD86 on the surface,significantly higher secretion of IL-12 and proliferation of lymphocytes than DCs with other treatments;The supernatants of MFC could induced the damage of DCs.Increased level of survival rate and A20 protein in the union group of DCs were found.CONCLUSION OK-432 in a combination with Poly(I:C)can effectively induce maturation DCs;OK-432 in a combination with Poly(I:C)can enhance the survival rate of DCs,which is associated with the increased expression of A20 protein.

关 键 词：树突状细胞 OK-432 聚肌胞 锌指蛋白A20 小鼠前胃癌 

分 类 号：R965[医药卫生—药理学]