壮药透骨香抗类风湿关节炎药物活性部位筛选及机制研究
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摘要
目的探讨壮药透骨香抗类风湿关节炎作用的药物活性部位及作用机制。方法体外培养法制备BALB/c小鼠脾淋巴细胞,经刀豆蛋白A(Co A)刺激作用后,以透骨香各药物部位进行干预,细胞计数试剂盒-8(CCK-8)法检测细胞增殖情况,计算刺激指数;体外培养小鼠脾细胞,刀豆蛋白A刺激T淋巴细胞增殖,采用酶联免疫吸附实验(ELISA)法检测观察透骨香活性部位对细胞因子白细胞介素-2(IL-2)、白细胞介素-10(IL-10)含量的影响。结果小鼠脾细胞经刀豆蛋白A刺激后,细胞计数试剂盒-8检测光密度(OD)值与刺激指数显著升高(P<0.05),透骨香正丁醇部位及水溶部位能明显降低光密度值及刺激指数,其中水溶部位(5μg·m L-1)作用最强(P<0.05);刀豆蛋白A刺激脾淋巴细胞增殖后,细胞因子白细胞介素-2、白细胞介素-10分泌显著增强,透骨香水溶部位干预后能明显抑制白细胞介素-2分泌,提高白细胞介素-10含量(P<0.05)。结论透骨香水溶部位对T淋巴细胞增殖具有较强抑制作用,是透骨香抗类风湿关节炎活性部位,其机制可能与调节细胞因子白细胞介素-2、白细胞介素-10分泌有关。
Objective To explore the anti-rheumatoid arthritis component of Zhuang medicine Alyxia levinei Merr.and its mechanism.Methods Splenic lymphocytes of BALB/c mice were cultured in vitro,which were intervened by the different extraction from Alyxia levinei Merr. after the stimulation of Concanavalin A( Co A).Cell proliferation were determined by CCK-8 to calculate stimulation index.The effect of Alyxia levinei Merr.component on IL-2,IL-10 was measured by ELISA after spleen cell of mice were cultured in vitro and the proliferation of T-lymphacye were stimulated by Co A.Results Optical density( OD) and stimulation index of splenocyte,determined by CCK-8,were significantly increased( P< 0.05) after stimulation by Co A.The butanol and water extract of Alyxia levinei Merr.can obviously decrease the OD and stimulation index,especially the water extract( 5 μg·m L-1)( P<0.05).After proliferation of splenic lymphocytes which was stimulated by Co A,excretion of IL-2 and IL-10 enhanced obviously.IL-2 was inhibited and the IL-10 was improved greatly by the water extract of Alyxia levinei Merr.( P<0.05).Conclusion The water extract of Alyxia levinei Merr.is the bioactive component of anti-rheumatoid arthritis which has potent inhibitory effects on T-lymphocytes proliferation.Its mechanism may be related to the regulation of IL-2 and IL-10.
Objective To explore the anti-rheumatoid arthritis component of Zhuang medicine Alyxia levinei Merr.and its mechanism.Methods Splenic lymphocytes of BALB/c mice were cultured in vitro,which were intervened by the different extraction from Alyxia levinei Merr. after the stimulation of Concanavalin A( Co A).Cell proliferation were determined by CCK-8 to calculate stimulation index.The effect of Alyxia levinei Merr.component on IL-2,IL-10 was measured by ELISA after spleen cell of mice were cultured in vitro and the proliferation of T-lymphacye were stimulated by Co A.Results Optical density( OD) and stimulation index of splenocyte,determined by CCK-8,were significantly increased( P< 0.05) after stimulation by Co A.The butanol and water extract of Alyxia levinei Merr.can obviously decrease the OD and stimulation index,especially the water extract( 5 μg·m L-1)( P<0.05).After proliferation of splenic lymphocytes which was stimulated by Co A,excretion of IL-2 and IL-10 enhanced obviously.IL-2 was inhibited and the IL-10 was improved greatly by the water extract of Alyxia levinei Merr.( P<0.05).Conclusion The water extract of Alyxia levinei Merr.is the bioactive component of anti-rheumatoid arthritis which has potent inhibitory effects on T-lymphocytes proliferation.Its mechanism may be related to the regulation of IL-2 and IL-10.
引文
[1]李凌汉,麦培根,陈宝红.雷公藤多甙联合免疫抑制剂治疗类风湿关节炎疗效及对炎性因子的影响[J].现代中西医结合杂志,2017,26(10):1088-1090.
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[8]戎瑞雪,王中傲,李晓婵,等.绒白乳菇中提取的乳菇菌素-D的免疫抑制作用[J].科学技术与工程,2018,18(1):187-191.
[9] MARIASELVAM C M,AOKI M,SALAH S,et al.Cytokineexpression and cytokine-based T cell profiling in SouthIndian rheumatoid arthritis[J]. Immunobiology,2014,219(10):772-777.
[10]孟岩,李鸣远,张新玉,等.类风湿关节炎患者血清TNF-α、IL-2及IL-13变化的价值研究[J].海南医学院学报,2018,24(2):188-190,194.
[11]杨荣敏,段相国,陈建,等.血清IL-2、TNF-α及IL-13在类风湿关节炎诊断和治疗中的临床意义[J].现代免疫学,2017,37(1):44-49.
[12] XYSTRAKIS E,BOSWELL S E,HAWRYLOWICZ C M.Tregulatory cells and the control of allergic disease[J].Ex-pert Opin Biol Ther,2006,6(2):121-133.
[13]周大兵,王瑞林,徐瑾,等.类风湿关节炎滑膜组织中IL-17和IL-10的表达及意义[J].临床与实验病理学杂志,2014,30(5):527-530.
[2] PRADHAN P,BHARDWAJ A,VENKATACHALAM V P.Bilateral cricoarytenoid arthritis:a cause of recurrentupper airway obstruction in rheumatoid arthritis[J].Malays J Med Sci,2016,23(3):89-91.
[3]张颖,宋志钊,刘元,等.壮药透骨香的质量标准研究[J].中国民族民间医药,2017,26(19):9-11.
[4]庞宇舟,林辰.实用壮医内科学[M].南宁:广西科学技术出版社,2011:202.
[5]赵玲,胡耀华,柳润辉,等.中药治疗类风湿性关节炎及临床应用的研究进展[J].药学实践杂志,2011,29(6):416-420,425.
[6]黄安,徐晶,陈筱悦,等.类风湿关节炎的壮医治疗研究进展[J].中国中医基础医学杂志,2016,22(8):1144-1146.
[7]李巧霞,丛斌,单保恩.T细胞与类风湿性关节炎[J].国际免疫学杂志,2011,34(1):1-5.
[8]戎瑞雪,王中傲,李晓婵,等.绒白乳菇中提取的乳菇菌素-D的免疫抑制作用[J].科学技术与工程,2018,18(1):187-191.
[9] MARIASELVAM C M,AOKI M,SALAH S,et al.Cytokineexpression and cytokine-based T cell profiling in SouthIndian rheumatoid arthritis[J]. Immunobiology,2014,219(10):772-777.
[10]孟岩,李鸣远,张新玉,等.类风湿关节炎患者血清TNF-α、IL-2及IL-13变化的价值研究[J].海南医学院学报,2018,24(2):188-190,194.
[11]杨荣敏,段相国,陈建,等.血清IL-2、TNF-α及IL-13在类风湿关节炎诊断和治疗中的临床意义[J].现代免疫学,2017,37(1):44-49.
[12] XYSTRAKIS E,BOSWELL S E,HAWRYLOWICZ C M.Tregulatory cells and the control of allergic disease[J].Ex-pert Opin Biol Ther,2006,6(2):121-133.
[13]周大兵,王瑞林,徐瑾,等.类风湿关节炎滑膜组织中IL-17和IL-10的表达及意义[J].临床与实验病理学杂志,2014,30(5):527-530.