喂食蛹虫草的小鼠血清对小鼠脾淋巴细胞增殖及活化的影响
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摘要
探讨喂食蛹虫草(Cordyceps militaris)的小鼠血清对脾淋巴细胞的影响。将20只雄性BALB/c小鼠分为两组。蛹虫草组用蛹虫草水提物(10 mL/kg·d),对照组用等体积纯净水,连续灌胃30 d。收集上述两组小鼠血清,通过小鼠脾淋巴细胞体外增殖实验检测不同浓度(5%、10%、20%)血清对小鼠脾淋巴细胞增殖和活化的影响。采用流式细胞术检测T细胞(CD~(3+))、B细胞(CD~(19+))、辅助性T细胞(helper T,Th)(CD~(3+)CD~(4+))、细胞毒性T细胞(cytotoxic T cell,Tc)(CD~(3+)CD~(8+))在淋巴细胞中所占百分比,以及细胞表面共刺激因子CD28和早期活化蛋白CD69的表达量。采用酶联免疫吸附法测定小鼠脾淋巴细胞上清液中干扰素γ(interferon-γ,IFN-γ)和白细胞介素-2(interleukin-2,IL-2)的表达量。结果表明:与对照组相比,培养基中添加10%和20%喂食蛹虫草的小鼠血清可刺激脾淋巴细胞增殖,刺激指数分别为(105.25±2.40)%和(134.54±5.99)%;喂食蛹虫草的小鼠血清可明显提高小鼠脾淋巴细胞中T细胞和B细胞的百分比,与对照组相比,添加10%和20%喂食蛹虫草的小鼠血清使T细胞在淋巴细胞总数中的百分比分别提高了(3.00±0.20)%和(7.20±0.21)%,添加20%喂食蛹虫草的小鼠血清使B细胞的百分比提高了(8.53±0.25)%;喂食蛹虫草的小鼠血清可促进Th细胞、Tc细胞分化,上调T细胞及其亚群中CD28分子的表达,提高T细胞和B细胞中CD69分子的表达,促进淋巴细胞活化,还可显著促进小鼠脾淋巴细胞中免疫调节因子IFN-γ和IL-2的表达;添加20%喂食蛹虫草的小鼠血清IFN-γ表达量为(101.92±12.76)ng/L,添加10%和20%喂食蛹虫草的小鼠血清IL-2表达量分别为(369.56±30.98)ng/L和(417.69±15.94)ng/L。
To investigate the effect of serum from mice fed with Cordyceps militaris(SMFCM)on splenic lymphocytes,twenty male BALB/c mice were divided into Cordyceps militaris group and control group.Cordyceps militaris group was treated with aqueous extract from Cordyceps militaris(10 mL/kg·d by gavage),while the control group was given distilled water of equal volume for 30 days.Sera were collected from the above two groups.The effect of serum at different concentrations(5%,10% and 20%)on proliferation and activation of mouse splenic lymphocytes were detected by in vitro proliferation test.The percentages of T cells(CD~(3+)),B cells(CD~(19+)),helper T cells(Th,CD~(3+)CD~(4+))and cytotoxic T cells(Tc,CD~(3+)CD~(8+)),and the expression of costimulatory factor CD28 and activation characteristic protein CD69 on the surface of these cells were detected by flow cytometry.The concentrations of interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of mouse splenic lymphocytes were determined by enzyme-linked immunosorbent assay(ELISA).The results showed that 10% and 20% addition of SMFCM to medium stimulated the proliferation of splenic lymphocytes,and the stimulation indices were 105.25±2.40% and134.54±5.99 % respectively compared to control.SMFCM significantly increased the percentages of T cells and B cells in mouse splenic lymphocytes.In comparison to the control group,the percentage of T cells in the total lymphocyte count increased by 3.00±0.210% and 7.20±0.21 % when treated with 10 % and 20% of SMFCM in medium respectively.The percentages of B cells increased by 8.53±0.25 % in the 20% SMFCM group.SMFCM promoted the differentiation of Th cells and Tc cells,up-regulated the expression of CD28 in T cells and their subsets,elevated the expression of CD69 in T cells and B cells,encouraged lymphocyte activation,and significantly improved the expression of IFN-γ and IL-2 in mouse spleen lymphocytes.IFN-γlevel was 101.92±12.76 ng/L when treated with 20% of SMFCM,and IL-2 levels were 369.56±30.98 ng/L and 417.69±15.94 ng/L when treated with 10% and 20% of SMFCM at respectively.
To investigate the effect of serum from mice fed with Cordyceps militaris(SMFCM)on splenic lymphocytes,twenty male BALB/c mice were divided into Cordyceps militaris group and control group.Cordyceps militaris group was treated with aqueous extract from Cordyceps militaris(10 mL/kg·d by gavage),while the control group was given distilled water of equal volume for 30 days.Sera were collected from the above two groups.The effect of serum at different concentrations(5%,10% and 20%)on proliferation and activation of mouse splenic lymphocytes were detected by in vitro proliferation test.The percentages of T cells(CD~(3+)),B cells(CD~(19+)),helper T cells(Th,CD~(3+)CD~(4+))and cytotoxic T cells(Tc,CD~(3+)CD~(8+)),and the expression of costimulatory factor CD28 and activation characteristic protein CD69 on the surface of these cells were detected by flow cytometry.The concentrations of interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of mouse splenic lymphocytes were determined by enzyme-linked immunosorbent assay(ELISA).The results showed that 10% and 20% addition of SMFCM to medium stimulated the proliferation of splenic lymphocytes,and the stimulation indices were 105.25±2.40% and134.54±5.99 % respectively compared to control.SMFCM significantly increased the percentages of T cells and B cells in mouse splenic lymphocytes.In comparison to the control group,the percentage of T cells in the total lymphocyte count increased by 3.00±0.210% and 7.20±0.21 % when treated with 10 % and 20% of SMFCM in medium respectively.The percentages of B cells increased by 8.53±0.25 % in the 20% SMFCM group.SMFCM promoted the differentiation of Th cells and Tc cells,up-regulated the expression of CD28 in T cells and their subsets,elevated the expression of CD69 in T cells and B cells,encouraged lymphocyte activation,and significantly improved the expression of IFN-γ and IL-2 in mouse spleen lymphocytes.IFN-γlevel was 101.92±12.76 ng/L when treated with 20% of SMFCM,and IL-2 levels were 369.56±30.98 ng/L and 417.69±15.94 ng/L when treated with 10% and 20% of SMFCM at respectively.
引文
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[14]高青,龙军,简立信,等.复方蛹虫草颗粒对小鼠免疫功能以及脾淋巴细胞增殖的影响[J].中国实验方剂学杂志,2013,19(16):259-263.
[15]杨磊,黄洋,龙珍.蛹虫草多糖抑制刀豆蛋白诱导的T细胞增殖[J].贵州医药,2017,41(8):810-813.
[16]谭政,龚非力.T细胞功能亚群[J].生命科学,2010,22(6):499-505.
[17]LAHN M,KALATARADI H,MITTELSTADT P, et al.Early preferential stimulation of gamma delta T cells by TNF-alpha[J].Journal of Immunology,1998,160(11):5221-5230.
[18]王平.T细胞表面分子CD28研究进展[J].国外医学免疫学分册,1995(3):113-116.
[19]吕欢,万龙,姜明燕.复方HD对环磷酰胺致免疫抑制小鼠免疫功能的影响[J].药物评价研究,2018,41(4):577-581.
[20]陈健明,黄健英,秦璐,等.黄芪皂苷对不同证型C57BL/6小鼠血清IL-2、IL-4、IgG1、IgG2水平的影响[J].儿科药学杂志,2018,24(8):1-4.
[2]郑壮丽,黄春花,梅彩英,等.蛹虫草国内外研究的新进展[J].环境昆虫学报,2011,33(2):225-233.
[3]孟泽彬,陈林会,韩近雨,等.蛹虫草化学活性成分的研究进展[J].分子植物育种,2015,13(9):2147-2154.
[4]余伯成,唐永范,唐亮,等.虫草素的药理作用研究进展[J].现代药物与临床,2011,26(5):349-352.
[5]陈春娟,朱振元,陈璐.低分子量蛹虫草多糖降血糖活性的研究[J].现代食品科技,2017,33(4):25-30.
[6]NAKAMURA K,KONOHA K,YAMAGUCHI Y,et al.Combined effects of Cordyceps sinensis and methotrexate on hematogenic lung metastasis in mice[J].Receptors Channels,2003,9(5):329-334.
[7]黄小莉,朱培欣,胡远亮,等.蛹虫草多糖对急性酒精性肝损伤改善作用的研究[J].菌物学报,2017,36(2):242-250.
[8]朱振元,刘瑶,薛春茂.蛹虫草色素提取及其抗氧化活性的研究[J].食品科技,2016,41(6):236-240.
[9]王米,张丽芳,费陈忠,等.蛹虫草多糖对小鼠腹腔巨噬细胞免疫功能的影响[J].中国生化药物杂志,2015,35(4):10-12.
[10]张圣方,赵龙玉,赵凤春,等.泰山蛹虫草多糖对免疫抑制小鼠肠道菌群及分泌型免疫球蛋白A的影响[J].食品科学,2015,36(5):148-152.
[11]樊慧婷,林洪生.蛹虫草化学成分及药理作用研究进展[J].中国中药杂志,2013,38(15):2549-2552.
[12]刘春发,胡建新,屈新辉.中药制剂对免疫功能促进作用的研究进展[J].中国医药导报,2013,10(28):27-33.
[13]王米,张丽芳,杨锐乐,等.蛹虫草多糖对小鼠脾淋巴细胞增殖及IL-2和IFN-γ分泌的影响[J].中国兽医科学,2015,45(8):848-853.
[14]高青,龙军,简立信,等.复方蛹虫草颗粒对小鼠免疫功能以及脾淋巴细胞增殖的影响[J].中国实验方剂学杂志,2013,19(16):259-263.
[15]杨磊,黄洋,龙珍.蛹虫草多糖抑制刀豆蛋白诱导的T细胞增殖[J].贵州医药,2017,41(8):810-813.
[16]谭政,龚非力.T细胞功能亚群[J].生命科学,2010,22(6):499-505.
[17]LAHN M,KALATARADI H,MITTELSTADT P, et al.Early preferential stimulation of gamma delta T cells by TNF-alpha[J].Journal of Immunology,1998,160(11):5221-5230.
[18]王平.T细胞表面分子CD28研究进展[J].国外医学免疫学分册,1995(3):113-116.
[19]吕欢,万龙,姜明燕.复方HD对环磷酰胺致免疫抑制小鼠免疫功能的影响[J].药物评价研究,2018,41(4):577-581.
[20]陈健明,黄健英,秦璐,等.黄芪皂苷对不同证型C57BL/6小鼠血清IL-2、IL-4、IgG1、IgG2水平的影响[J].儿科药学杂志,2018,24(8):1-4.
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