黄芪皂苷Ⅱ调控CD45 PTPase诱导抗肿瘤免疫效应的研究  

Anti-tumor Immunological Mechanism of Astragaloside Ⅱ through Activating CD45 PTPase

作　　者：王敏[1] 郑喜 普晓佳 万春平 WANG Min;ZHENG Xi;PU XiaoJiao;WAN Chunping(Department of Emergency Medicine,the First People's Hospital of Yunnan Province,Kunming 650021,China;The Central Laboratory,The First Affiliated Hospital of Yunnan University of Chinese Medicine,Kunming 650021,China;College of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南省第一人民医院急症医学部,云南昆明650032 [2]云南中医药大学第一附属医院,云南昆明650021 [3]云南中医药大学中药学院,云南昆明650500

出　　处：《云南中医学院学报》2021年第2期1-6,共6页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81460624,81960745);云南省自然基金面上项目(2015FB199);云南省科技厅-云南中医学院应用基础研究联合专项〔2017FF117(-041)。

摘　　要：目的 探讨黄芪皂苷Ⅱ(Astragaloside Ⅱ,ASI Ⅱ)调控CD45蛋白酪氨酸磷酸酯酶(CD45 PTPase)诱导抗肿瘤免疫效应,为黄芪临床应用于抗肿瘤治疗提供参考。方法复制H22荷瘤肿瘤小鼠模型,分为模型组、ASI Ⅱ组和ASI Ⅱ+抗CD45抗体组。药物干预21d后,测定瘤重和瘤径;MTT法测定模型各组小鼠脾淋巴细胞增殖活力;qPCR评价各组脾淋巴细胞Th1细胞相关因子的转录水平。流式细胞术检测脾淋巴细胞Th1(CD4+IFN-γ+T)细胞和Treg(CD4+Foxp3+)细胞表达水平。结果与模型组比较,ASI Ⅱ组瘤径、瘤重和瘤积均低于模型组(P<0.05);ASI Ⅱ组脾淋巴细胞增殖活力(OD值)显著高于模型组,差异具有统计学意义(P<0.05)。qPCR检测结果显示,ASI Ⅱ组小鼠脾淋巴细胞Th1细胞因子(IL-2和IFN-γ)及特异转录因子T-bet和Th2细胞因子(IL-4)mRNA表达显著高于模型(均为P<0.05);流式细胞术检测结果显示,ASI Ⅱ组脾淋巴细胞CD4+IFN-γ+(Th1细胞)比例显著高于模型组(P<0.05),而CD4+Foxp3+(Treg细胞)比例低于模型组(P<0.05)。ASI Ⅱ+CD45抗体组抗肿瘤效应、淋巴细胞的增殖活力、细胞因子L-2、IFN-γ和IL-4 mRNA表达均高于ASI Ⅱ组;CD45抗体干预,减少了黄芪皂苷Ⅱ促IFN-γ细胞比例升高(P<0.05),对ASI Ⅱ诱导Treg细胞下调无明显的影响。结论黄芪皂苷Ⅱ可能激活CD45 PTPase,诱导Th1细胞反应,增强抗肿瘤免疫反应。Objective This paper was designed to assess the anti-tumor immunological effect and reveal the molecular mechanism of Astragaloside Ⅱ in enhancing the anti-tumor immunological response via regulating CD45 PTPase.This study will provide a basis for the theoretical foundation of antitumor immunotherapy of Astragalus. Methods The H22 tumor-bearing mice was established to investigate the anti-tumor immunological effect of Astragaloside Ⅱ. Mice were divided into three groups, including model group, Astragaloside Ⅱ group, and Astragaloside Ⅱ+Anti-CD45 Ab group. After21 days of intragastric administration, the tumor tissue was isolated to assess tumor weight, tumor diameter and tumor volume. The lymphocyte cells proliferation from H22 tumor-bearing mice was detected by MTT method. The mRNA expression of Th1 cytokine, Th2 cytokine, and transcript factor T-bet were examined by q-PCR analysis. The expression of Th1(CD4+IFN-γ+), Treg(CD4+Foxp3+) and Th17(CD4+IL-17+) were detected by flow cytometry. Results The tumor weight, tumor diameter and tumor volume in Astragaloside Ⅱ group are more small than model group( P<0.05). Astragaloside Ⅱ treatment significantly enhance the lymphocyte proliferation activity from H22 tumor-bearing mice in response to ConA,the transcriptional expression in Th1 and Th2, including IFN-γ, IL-2, IL-4, and T-bet were up-regulated. Meanwhile, IFN-γexpression was up-regulated in CD4+T cells but down-regulated in T regulatory cells By Astragaloside Ⅱ. Furthermore, anti-mouse CD45 Ab treatment intensely blocked the anti-tumor effect and lymphocyte proliferation activity which induced by Astragaloside. Compared with Astragaloside Ⅱ group, IL-2, IFN-γ and IL-4 mRNA expression was decreased in Astragaloside Ⅱ+Anti-CD45 Ab group. Anti-mouse CD45 Ab treatment intensely down-regulated IFN-γ expression in CD4+T cells without significantly influencing Treg cells. Conclusion Astragaloside Ⅱ might enhanced anti-tumor immunological response via regulating CD45 protein tyrosine phosphatase

关 键 词：黄芪皂苷Ⅱ Γ-干扰素 CD45 PTPase TH1细胞 

分 类 号：R285.5[医药卫生—中药学]