大黄附子汤增强NK细胞杀伤小鼠LLC细胞效力  

Dahuang Fuzi decoction enhances killing effect of NK cells on Lewis lung carcinoma cells

作　　者：王晓燕[1] 王茗萱 冯嘉轩 白金萍[1] 王冰梅[1] 刘畅[1] WANG Xiaoyan;WANG Mingxuan;FENG Jiaxuan;BAI Jinping;WANG Bingmei;LIU Chang(Clinical Medical College,Changchun University of Traditional Chinese Medicine,Changchun 130021,China)

机构地区：[1]长春中医药大学临床医学学院,吉林长春130021

出　　处：《中国兽医学报》2024年第7期1483-1488,共6页Chinese Journal of Veterinary Science

基　　金：吉林省教育厅“十三五”科学技术资助项目(JJKH20190470KJ);吉林省中医药管理局科技资助项目(2019179);国家级大学生创新创业训练计划资助项目(202210199029);吉林省级大学生创新创业训练计划资助项目(S202310199064)。

摘　　要：旨在探讨大黄附子汤对自然杀伤细胞(NK细胞)杀伤效力的调节作用及其小鼠肺癌移植瘤生长的影响。构建Lewis小鼠肺癌移植瘤模型,试验随机分为生理盐水、大黄附子汤低、高浓度组,检测大黄附子汤低、高浓度治疗组对小鼠肺癌移植瘤生长的影响。通过ELISA检测大黄附子汤对IFN-γ释放量以及IL-2、IL-10水平的影响。通过钙黄绿素释放试验,检测NK细胞的杀伤效力。通过流式细胞术检测CD107α的释放量;检测对照组及治疗组活化性受体表达及肿瘤细胞表面配体的表达。结果显示,大黄附子汤治疗组以剂量依赖方式抑制Lewis小鼠移植瘤生长(P<0.05)。ELISA检测肿瘤组织中IFN-γ、IL-2水平增加(P<0.05),IL-10变化不明显。钙黄绿素释放试验检测表明治疗组NK细胞的杀伤效率增加(P<0.05)。ELISA检测培养上清中IFN-γ的分泌水平增加(P<0.05)。通过流式细胞仪检测发现大黄附子汤提高NK细胞CD107α表达水平增加;上调A549、H1299细胞表面MIC A/B表达,但没有上调抑制性配体HLA-ABC表达。表明大黄附子汤通过上调MIC A/B的表达,增加IFN-γ和CD107α分泌,增强NK细胞杀伤小鼠肺癌细胞效应。综上所述,大黄附子汤抑制肺癌移植瘤生长可能是通过增强NK细胞对肺癌细胞的杀伤作用。The aim of this study is to investigate the effect of Dahuang Fuzi decoction(DHFZT)on the killing effect of natural killer(NK)cells and the growth of lung cancer xenografts in mice.Lewis lung cancer xenograft model was constructed,and the mice were randomly divided to the normal saline group,low and high concentrations of DHFZT groups.The growth of lung cancer xenografts in mice in the control group,low and high concentration of DHFZT treatment groups was detected.The levels of IFN-γ,IL-2and IL-10were detected by ELISA.The killing effect of NK cells was detected by calcein release assay.The release of CD107αwas detected by flow cytometry.The expression of activated receptors and the tumor cell surface ligands were detected in the control group and DHFZT treatment groups.The results showed that the xenografts of Lewis mice in the DHFZT treatment groups were grown slower than that in the control group in a dose dependent manner(P<0.05).Compared with the control groups,the levels of IL-2and IFN-γin tumor tissues were increased in DHFZT treatment groups(P<0.05),but no significant changes in IL-10.Calcein release assay showed that the killing efficiency of NK cells in the DHFZT treatment groups was increased(P<0.05).The secretion level of IFN-γin the culture supernatant was increased by DHFZT treatment(P<0.05).DHFZT treatment increased the expression level of CD107αof NK cells.The expression of MIC A/B on the surface of A549and H1299cells was upregulated by DHFZT treatment,but not the expression of inhibitory ligand HLA-ABC.The results showed that DHFZT enhanced the killing effect of NK cells on mouse lung cancer cells by up-regulating the expression of MIC A/B,increasing the secretion of IFN-γand CD107α.Hence,these results indicated that DHFZT suppresses the lung cancer growth potential through regulating the killing effect of NK cells.

关 键 词：大黄附子汤 NK细胞 肺癌 MIC A/B 小鼠 

分 类 号：R734.2[医药卫生—肿瘤] R392.5[医药卫生—临床医学]