苯并噁唑衍生物PO-296对树突状细胞分化及相关指标的影响
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篇名: 苯并噁唑衍生物PO-296对树突状细胞分化及相关指标的影响
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摘要: 目的:探讨苯并噁唑衍生物2-(氯苯并噁唑基-2基)-4,5,6,7-四氢-二氢-吲唑-3-醇(PO-296)对小鼠骨髓来源树突状细胞(DC)分化及其特异性表面分子、炎症细胞因子等相关指标的影响。方法:分离小鼠骨髓核细胞,以重组小鼠粒细胞巨噬细胞集落刺激因子和重组小鼠白细胞介素4刺激获取未成熟DC(imDC);经低、中、高剂量(1、5、25 μmol/L)PO-296预处理后,以脂多糖诱导获取DC。应用流式细胞术检测DC特异性表面分子[即Ⅱ类主要组织相容性复合物(MHC Ⅱ)、CD80、CD86和趋化因子受体7(CCR7)阳性细胞比例]的表达以及imDC吞噬能力(即葡聚糖阳性细胞比例)和DC存活情况(即存活细胞比例),采用酶联免疫吸附测定法检测DC培养液中炎症细胞因子[白细胞介素10(IL-10)、IL-12、肿瘤坏死因子α(TNF-α)]水平。结果:与imDC组比较,空载组MHC Ⅱ、CD80、CD86阳性细胞比例均显著升高(P<0.05)。与空载组比较,PO-296各剂量组细胞培养液中IL-10水平均显著升高,阳性组以及PO-296中、高剂量组细胞MHC Ⅱ、CD80、CD86阳性细胞比例以及各给药组细胞培养液中IL-12、TNF-α水平均显著降低(P<0.05);而各给药组CCR7阳性细胞、葡聚糖阳性细胞和存活细胞比例与空载组比较,差异均无统计学意义(P>0.05)。结论:PO-296无明显的细胞毒性,亦不影响imDC的吞噬功能;同时,该化合物可抑制DC特异性表面分子的表达,调控其炎症细胞因子的分泌。
ABSTRACT: OBJECTIVE: To investigate the effects of benzoxazole derivative 2-(chlorobenzoxazolyl-2-yl)-4,5,6,7-tetrahydro- dihydro-indazole-3-ol (PO-296) on the differentiation of murine bone marrow-derived dendritic cells(DC) and their related indexes as specific surface molecules and inflammatory cytokines. METHODS: Bone marrow nuclear cells of mice were isolated, and immature DC (imDC) was obtained by recombinant mice granulocyte macrophage colony-stimulating factor and recombinant mice IL-4. After pretreated with low-dose, medium-dose and high-dose (1, 5, 25 μmol/L) of PO-296, DC was obtained by lipopolysaccharide induction. Flow cytometry was used to detect the expression of DC specific surface molecules [i.e. the proportion of class Ⅱ major histocompatibility complex (MHC Ⅱ), CD80, CD86 and chemokine receptor 7 (CCR7) positive cells], imDC phagocytosis (i.e. the proportion of dextran positive cells) and DC survival (i.e. the proportion of survival cells). ELISA method was used to detect the levels of inflammatory cytokines (IL-10, IL-12 and TNF-α) in cell culture medium. RESULTS: Compared with imDC group, the proportion of MHC Ⅱ, CD80 and CD86 positive cells were increased significantly in non-loading group (P<0.05). Compared with non-loading group, the levels of IL-10 in cell culture medium were increased significantly in PO-296 groups. The proportions of MHC Ⅱ, CD80 and CD86 positive cells in positive group and PO-296 medium-dose and high-dose groups as well as the levels of IL-12 and TNF-α in cell culture medium in administration groups were decreased significantly (P<0.05). There was no statistical significance in the proportion of CCR7 positive cells, dextran positive cells and survival cells in administration groups, compared with non-loading group (P>0.05). CONCLUSIONS: PO-296 has no obvious cytotoxicity and does not affect the phagocytic function of imDC. At the same time, the compound can inhibit the expression of DC specific surface molecules and regulate the secretion of inflammatory cytokines.
期刊: 2019年第30卷第18期
作者: 朱薪颖,杨苗,徐灿,彭旭,段佳毅,刘丹,王衍堂
AUTHORS: ZHU Xinying,YANG Miao,XU Can,PENG Xu,DUAN Jiayi,LIU Dan,WANG Yantang
关键字: PO-296;树突状细胞;特异性表面分子;炎症细胞因子;吞噬功能;细胞毒性
KEYWORDS: PO-296; Dendritic cell; Specific surface molecules; Inflammatory cytokines; Phagocytic function; Cytotoxicity
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