氧化苦参碱对淋巴细胞增殖和调节性T细胞（Tr）数量的影响

目的： 分析氧化苦参碱（OMT）对小鼠外周血调节性T细胞（Tr细胞）数量及其对刀豆蛋白A(Con A) 刺激的小鼠淋巴结T细胞增殖的影响, 探讨OMT治疗ACD的免疫学机制。方法： 建立DNFB诱发的小鼠ACD模型，腹腔注射（ip）不同剂量的OMT、PBS、氢化可的松(HCT)，在实验的第1 d、7 d、14 d、21 d、28 d小鼠尾静脉采血, 应用抗-CD3、抗-CD4、抗-CD25单抗进行免疫荧光标记, 流式细胞术检测各组CD4+CD25+T细胞数量。利用羧基荧光素乙酰乙酸(CFDA-SE)染色，流式细胞术检测OMT对小鼠淋巴结T细胞增殖的影响。结果： 500、125和31 mg/L OMT对小鼠淋巴结T细胞增殖呈剂量依赖性抑制，而16、8、4、2 mg/L OMT可促进小鼠淋巴结T细胞增殖，但剂量依赖关系不明显。腹腔注射OMT能明显提高小鼠外周血中CD4+CD25+T细胞数量, 与HCT组、PBS组比较（P<0.01）。结论: OMT对小鼠淋巴结T细胞增殖呈双向作用；腹腔注射OMT能明显提高小鼠外周血中CD4+CD25+T细胞数量；提示:OMT是一种双向免疫调节剂。

Effects of oxymatrine on lymphocyte proliferation and the quantity of regulatory T cells

AIM:To analyze the effects of oxymatrine(OMT) on the quantity of murine regulatory T cells(Tr cells) in the peripheral blood and mouse lymphocyte proliferation stimulated by Con A,and to probe into the immunological mechanism that OMT treats allergic contact dermatitis(ACD).METHODS:An ACD mouse model stimulated by dinitrofluorobenzene(DNFB) was established.Different dosages of OMT,PBS and hydrocortisone(HCT) were intraperitoneally injected(IP) into the mice.Blood samples were collected at 1 d,7 d,14 d,21 d and 28 d,then the T cells were isolated and marked with anti-CD3,anti-CD4,anti-CD25 three-colored immune fluorescence antibody to detect the quantity of CD4 CD25 T cells with flow cytometry.The fluorescence intensity changes of lymphocytes which were isolated from mouse's lymph node and co-stimulated by polyclonal stimulator Con A and OMT were examined by carboxyfluorescein diacetate succinimidyl ester(CFDA-SE) staining and flow cytometry.RESULTS:OMT at concentrations of 500,125 and 31 mg/L had the ability to restrain the proliferation of lymphocytes from lymph node in a dose dependent manner.However,OMT at concentrations of 16,8,4 and 2 mg/L promoted the proliferation of T lymphocytes from lymph node,but was not obviously dependent on its concentration.Intraperitoneal injection of OMT increased the numbers of CD4 CD25 T cell in peripheral blood obviously(P<0.01).CONCLUSION:The effects of OMT on the proliferation of T lymphocytes from mouse's lymph node cells are observed,OMT also increases the CD4 CD25 T cells in the peripheral blood,implying that OMT is a kind of immunoregulator with dual effects.