云南黑松露醇提物对小鼠T淋巴细胞免疫调节作用  

Regulatory Effect of Alcohol Extract of Yunnan Black Truffle on T Lymphocyte Immunity in Mice

作　　者：苏银芳 徐梓涵 袁雨欣 李干鹏[1] 祁艳艳 王芳 蔡正达 SU Yinfang;XU Zihan;YUAN Yuxin;LI Ganpeng;QI Yanyan;WANG Fang;CAI Zhengda(Key Laboratory of Chemistry in Ethnic Medicinal Resources,State Ethnic Affairs Commission&Ministry of Education of China,Yunnan Minzu University,Kunming 650500,China;Yunnan Institute of Science and Technology,Kunming 650500,China)

机构地区：[1]云南民族大学民族药资源化学国家民族事务委员会-教育部重点实验室,昆明650500 [2]云南省科学技术院,昆明650500

出　　处：《医药导报》2023年第11期1606-1614,共9页Herald of Medicine

摘　　要：目的观察云南黑松露醇提物对小鼠T淋巴细胞增殖及免疫功能的影响。方法将干燥云南黑松露粉末进行乙醇超声提取、乙酸乙酯萃取,水相及乙酸乙酯相部分样品分别采用C 18反相柱、凝胶柱进行分离、浓缩;噻唑蓝(MTT)法检测云南黑松露醇提物对刀豆蛋白A(ConA)刺激小鼠脾淋巴细胞增殖的影响;酶联免疫吸附测定(ELISA)法检测云南黑松露醇提物对Anti-CD3刺激小鼠脾淋巴细胞分泌γ干扰素(IFN-γ)和白细胞介素2(IL-2)的影响;蛋白印迹(Western blotting)法检测云南黑松露醇提物对小鼠脾淋巴细胞IFN-γ表达水平的影响;流式细胞术检测云南黑松露醇提物对小鼠脾淋巴细胞CD_(3)^(+)及CD_(4)^(+)细胞表达水平的影响。结果云南黑松露醇提物共分离得到组分17个。其中水相部分分离得到组分8个,记作1-8;乙酸乙酯相部分分离得到组分9个,记作A-I。MTT结果显示,在ConA诱导下,F号组分对小鼠脾淋巴细胞具有一定促增殖活性。ELISA结果显示,在Anti-CD3诱导下,1、F和G号组分可促进小鼠脾淋巴细胞IL-2和IFN-γ分泌。Western blotting结果显示,2、F和G号组分可提高小鼠脾淋巴细胞IFN-γ表达水平。流式细胞术检测结果显示,G号组分可促进小鼠脾淋巴细胞CD_(3)^(+)T细胞表达,1和2号组分可促进小鼠脾淋巴细胞CD_(4)^(+)T细胞表达。结论云南黑松露醇提物具有促进小鼠脾淋巴细胞增殖的活性成分,可激活T细胞分泌炎症因子IFN-γ和IL-2,提高IFN-γ及CD_(3)^(+)、CD_(4)^(+)细胞表达,具有一定免疫调节活性。Objective To investigate the effects of alcohol extract of Yunnan black truffle on T lymphocyte proliferation and immune function in mice.Methods The dried black truffle powder was extracted by ultrasonic ethanol extraction and ethyl acetate extraction.And some samples in the aqueous phase and ethyl acetate phase were separated and concentrated by C 18 reversed-phase column and gel column,respectively.The effect of black truffle alcohol extract on the proliferation of mouse spleen lymphocytes stimulated by concanavalin A(ConA)was detected by the MTT method.The effect of alcohol extract of black truffle on the secretion of interferon-γ(IFN-γ)and interleukin-2(IL-2)in spleen lymphocytes of mice stimulated by anti-CD3 was detected by enzyme-linked immunosorbent assay(ELISA).The effect of alcohol extract of black truffle on the expression of IFN-γin spleen lymphocytes was detected by Western blotting.The effect of alcohol extract of black truffle on the expression of CD_(3)^(+)and CD_(4)^(+)cells in spleen lymphocytes was detected by flow cytometry.Results A total of 17 components were obtained from the alcohol extract of black truffle,of which eight components were separated from the aqueous phase,and recorded as component 1 to 8.Nine components were obtained by partial separation of ethyl acetate,which was recorded as component A to I.The results of MTT showed that component F could promote the proliferation of mouse spleen lymphocytes induced by ConA.ELISA results showed that component 1,F,and G promoted the secretion of IL-2 and IFN-γin mouse spleen lymphocytes induced by anti-CD3.Western blotting results showed that component 2,F,and G increased the expression level of IFN-γin mouse spleen lymphocytes.The results of flow cytometry showed that component G promoted the expression level of CD_(3)^(+)T cells in mouse spleen lymphocytes,while component 1 and 2 promoted the expression level of CD_(4)^(+)T cells in spleen lymphocytes.Conclusion The alcohol extract of Yunnan black truffle can promote the proliferat

关 键 词：云南黑松露 脾淋巴细胞 Γ-干扰素 白细胞介素2 

分 类 号：R286[医药卫生—中药学] R965[医药卫生—中医学]