Tuftsin衍生物T肽的抗肿瘤作用及机制探讨  

Anti-tumor effect and mechanism of a tuftsin-derivative T peptide

作　　者：史振伟 罗东 孙琦 王聪聪 杨齐 马玥 杨飞[1,2] 黄转青 徐风华 SHI Zhenwei;LUO Dong;SUN Qi;WANG Congcong;YANG Qi;MA Yue;YANG Fei;HUANG Zhuanqing;XU Fenghua(Pharmaceutical Sciences Research Division,Department of Pharmacy,Medical Supply Center,Chinese PLA General Hospital,Beijing 100853,China;Chinese PLA Medical School,Beijing 100853,China)

机构地区：[1]解放军总医院医疗保障中心药剂科药学基础研究室,北京100853 [2]解放军医学院,北京100853

出　　处：《解放军医学院学报》2022年第2期211-218,共8页Academic Journal of Chinese PLA Medical School

摘　　要：背景生物反应调节剂促吞噬肽(Tuftsin)具有抗肿瘤作用,但体内半衰期极短,为此课题组合成了其衍生物T肽(T Peptide),但其抗肿瘤作用还不明确。目的 探讨Tuftsin衍生物T肽的抗肿瘤作用及其机制。方法 建立小鼠黑色素瘤移植瘤模型:C57BL/6小鼠12只,鼠龄6周,随机分为T肽(T Peptide)组和对照(Control)组,每组6只。两组小鼠右侧前肢腋下接种黑色素瘤细胞悬液0.1 m L,含B16-F10细胞5×105个。肿瘤细胞接种后当天(第0天)T肽组小鼠皮下注射T肽(药物剂量8 mg/kg),实验期间隔日给药,共计给药9次,共18 d^(+)对照组小鼠注射相同体积的0.9%氯化钠注射液,通过测定瘤重评价T肽对移植肿瘤生长的作用。利用流式细胞术(FACs)检测小鼠脾组织和移植瘤部位免疫细胞T淋巴细胞(CD3^(+)、CD3^(+)CD4^(+)、CD3^(+)CD8^(+)、CD3^(+)CD44^(+))、自然杀伤细胞(NK1.1^(+))、髓系来源抑制细胞(CD11b^(+)Gr-1^(+))、巨噬细胞(F4/80^(+)CD86^(+)、F4/80^(+)CD206^(+))的含量,采用酶联免疫吸附测定(ELISA)脾细胞培养上清液中的细胞因子、血清中细胞因子IL-2、IL-4、IL-10、IL-12、TNF-α、TGF-β、IFN-γ的表达情况。体外实验采用CCK-8法测定T肽和黑色素瘤B16-F10细胞共培养后对细胞生长的影响。结果 T肽在体外对肿瘤细胞增殖没有影响,但体内实验对小鼠黑色素瘤B16-F10细胞移植性肿瘤表现出抑制作用。与对照组相比,T肽治疗对肿瘤生长抑制率为56.45%,T肽组脾组织CD8^(+)T淋巴细胞、M1型巨噬细胞(F4/80^(+)CD86^(+))、CD3^(+)CD44^(+)T淋巴细胞、髓系来源抑制细胞(CD11b^(+)Gr-1^(+))含量显著增加,M2型巨噬细胞(F4/80^(+)CD206^(+))和自然杀伤细胞(NK1.1^(+))无明显变化^(+)与对照组相比,T肽组小鼠肿瘤部位CD3^(+)CD44^(+)T淋巴细胞的含量显著增加,而CD4^(+)T淋巴细胞、CD8^(+)T淋巴细胞、自然杀伤细胞(NK1.1^(+))、髓系来源抑制细胞(CD11b^(+)Gr-1^(+))、M1型巨噬细胞(F4/80^(+)CD86^(+))和M2�Background Tuftsin is an immunomodulator and has been certified to be able to inhibit tumor growth, whereas it is unstable in vivo with very short half-life period. T peptide is designed and developed as a new type of tuftsin derivative to extend its half-time, however, its anti-tumor effect and mechanism is still unclear. Objective To investigate the anti-tumor effect and mechanism of T peptide. Methods Female C57 BL/6 J mice aged 6–8 weeks were randomly divided into control group and T Peptide group with 6 mice in each group. A volume of 0.1 m L of B16-F10 cell suspension(5.0×106 cells/m L) was inoculated subcutaneously under the right forelimb armpit of C57 BL/6 J mice on day 0. The mice in the T Peptide group were administrated with T Peptide subcutaneously at a dose of 8 mg/kg very other day, while the control mice were treated by the vehicle solution(normal saline) in the same way. The effect of T peptide on the growth of transplanted tumor was evaluated by measuring tumor weight. Flow cytometry was used to detect T lymphocytes(CD3^(+), CD3^(+)CD4^(+), CD3^(+)CD8^(+), CD3^(+)CD44^(+)), natural killer cells(NK1.1^(+)), myeloidderived suppressor cells(CD11 b^(+)Gr-1^(+)) and macrophages(F4/80^(+)CD86^(+), F4/80^(+)CD206^(+)) both in mice spleens and at the transplanted tumor site. The expression of cytokines(IL-2, IL-4, IL-10, IL-12, TNF-α, TGF-β, IFN-γ) in the supernatant of spleen cells as well as in serum were determined by enzyme-linked immunosorbent assay. CCK-8 assay was employed to assess the in vitro cytotoxicity of T Peptide. Results T Peptide did not have any influence on tumor cell proliferation in vitro, but showed an inhibitory effect on subcutaneously transplanted melanoma B16-F10 tumor in vivo, with the inhibition rate being 56.45%. For T Peptide treated tumor-bearing mice, the level of CD3^(+)CD8^(+)T lymphocytes, M1 type macrophages(F4/80^(+)CD86^(+)), CD3^(+)CD44^(+)T lymphocytes, and myeloid-derived suppressor cells(CD11 b^(+)Gr-1^(+)) in the spleens increased significantly compared t

关 键 词：T肽 肿瘤抑制 免疫增强 小鼠黑色素瘤B16-F10细胞 促吞噬肽 

分 类 号：R965[医药卫生—药理学]