刘灏,吴小翎,王山梅,田洪军,昝利萍
LIU Hao,WU Xiaoling,WANG Shanmei,TIAN hongjun,ZAN Liping

• 1:重庆医科大学附属大学城医院消化中心
• 2:重庆医科大学附属第二医院消化内科
• 3:河南省人民医院检验科
• 4:重庆市西南铝加工厂职工医院内科

摘要(Abstract)：

目的:探讨再极化肿瘤巨噬细胞对胃癌生长的抑制作用。方法:70只雌性Balb/c小鼠分为体内实验和体外实验;体外实验分为9组,对照组、M2组、M1组,PM-LPS组、TPM-LPS组,PM-IL-12-LPS组、TPM-IL-12-LPS组,PM-IFN-γ-LPS组、TPM-IFN-γ-LPS组,用ELISA试剂盒检测各组巨噬细胞培养上清液中肿瘤坏死因子α(TNF-α)、白介素12p70(IL-12p70)和白介素10(IL-10),用QRT-PCT法检测巨噬细胞中诱导型一氧化氮合酶(iNOS)和精氨酸酶(Arg)mRNA的表达;体内实验分为6组,PM+MFC组、TPM+MFC组、M1+MFC组、M2+MFC组、IL-12-PM+MFC组和IL-12-TPM+MFC组,测量并比较各组MFC负荷瘤小鼠皮下移植瘤的体积与质量。结果:巨噬细胞上清液中,与对照组比较,M1、PM-IFN-γ-LPS、TPM-IFN-γ-LPS和TPM-IL-12-LPS组均高表达TNF-α和IL-12p70,低表达IL-10;M2和TPM-LPS组高表达IL-10,低表达TNF-α和IL-12p70(P<0.05);PM-IL-12-LPS与PM-LPS组比较,差异无统计学意义(P>0.05);LPS刺激后的TPM与M2组比较,差异无统计学意义(P>0.05);与对照组比较,M1组高表达iNOS mRNA,低表达Arg mRNA;M2组高表达Arg mRNA,低表达iNOS mRNA(P<0.05);TPM-LPS高表达Arg mRNA,低表达iNOS mRNA(P<0.05);M2+MFC组和TPM+MFC组小鼠肿瘤生长较快。接种后第28天,M2+MFC组、TPM+MFC组和PM+MFC组小鼠肿瘤体积和重量明显大于比IL-12-TPM+MFC组和M1+MFC组(P<0.05);M2组小鼠肿瘤最大,TPM组小鼠肿瘤大小仅次于M2组(P<0.05);与TPM组小鼠相比,IL-12再极化的TPM组小鼠肿瘤明显缩小(P<0.05)。结论:再极化巨噬细胞可作为肿瘤免疫治疗的一个新途径。
Objective: To explore the inhibitory effects of repolarized tumor macrophages on gastric tumor growth. Methods: Seventy female Balb /c mice served as study subject in vitro and in vivo.There were 9 groups in vitro,including control group,M2 group,M1 group,PM-LPS group,TPMLPS group,PM-IL-12-LPS group,TPM-IL-12-LPS group,PM-IFN-γ-LPS group and TPM-IFN-γ-LPS group. TNF-α,IL-12p70 and IL-10 levels in culture medium of macrophages were detected with ELISA kits,and mRNA expression levels of iNOS and arginase( Arg) were measured with qRT-PCR.There were 6 groups in vivo,including PM + MFC group,TPM + MFC group,M1 + MFC group,M2 +MFC group,IL-12-PM + MFC group and IL-12-TPM + MFC group. The volume and weight of subcutaneous transplantation tumor of mice were measured. Results: As compared with control group,TNF-αand IL-12p70 levels in M1 group,PM-IFN-γ-LPS group,TPM-IFN-γ-LPS group and TPM-IL-12-LPS group were increased and IL-10 levels were decreased( P < 0. 01),but TNF-α and IL-12p70 levels in M2 group and TPM-LPS group were decreased and IL-10 levels were increased( P < 0. 05). No statistical significance were found between PM-IL-12-LPS group and PM-LPS group( P > 0. 05),or between TPM-LPS group and M2 group( P > 0. 05). The level of iNOS mRNA in M1 group was higher than that in control group,but the level of Arg mRNA was lower than that in control group,whereas the levels of iNOS mRNA in both M2 group and TPM-LPS group were lower than those of control group,the levels of Arg mRNA were higher than those in control group( P < 0. 05). Tumor in mice of M2 + MFC group and TPM + MFC group grew rapidly. After subcutaneously implanted for 28 days,the volume and weight of the tumors in M2 + MFC group,TPM + MFC group and PM + MFC group were bigger and heavier than those in IL-12-TPM + MFC group and M1 + MFC group( P < 0. 05). Mice of M2 group had tumors with biggest volume and weight,and followed by TPM group( P < 0. 05). Compared with TPM group,the tumors of mice in IL-12-TPM group were obviously smaller( P < 0. 05). Conclusions: Repolarization tumor macrophages provide a new approach for tumor immunotherapy.

关键词(KeyWords)： 小鼠;胃肿瘤;巨噬细胞;肿瘤坏死因子α;白介素12p70;白介素10;一氧化氮合酶;精氨酸酶
mouse;gastric cancer;macrophage;tumor necrosis factor-α;interleukin 12p70;interleukin 10;nitric oxide synthase;arginase

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 刘灏,吴小翎,王山梅,田洪军,昝利萍
LIU Hao,WU Xiaoling,WANG Shanmei,TIAN hongjun,ZAN Liping

DOI: 10.19367/j.cnki.1000-2707.2014.05.021

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