妇科千金胶囊对小鼠脾淋巴细胞免疫活性的研究

Study on Immunological Activity of Fuke Qianjin Capsule to Mice's Splenic Lymphocytes

目的研究妇科千金胶囊对小鼠脾淋巴细胞体外增殖及免疫调节的影响。方法分离制备小鼠脾淋巴细胞,将细胞分为空白组(不做任何处理),刀豆蛋白A组(刀豆蛋白A干预,ConA组),LPS组(脂多糖干预,LPS组)及不同浓度的妇科千金胶囊组(FKQ),作用时间分别为24 h、48 h和72 h。CCK8法检测FKQ对小鼠脾淋巴细胞体外增殖的影响;MTT法检测ConA及LPS对脾淋巴细胞增殖的影响;流式细胞术检测各组小鼠脾淋巴细胞中CD4+和CD8+的比例并计算CD4+/CD8+的比值;ELISA检测各组细胞上清中IFN-γ、IL-2、IL-4和IL-10的表达。结果FKQ对小鼠脾淋巴细胞有显著的增殖作用,以浓度为30、40、50μg/mL时其作用越明显(P<0.01);FKQ能对ConA诱导的T淋巴细胞及LPS诱导的B淋巴细胞增殖有着明显的促进作用,以浓度为50μg/mL最显著(P<0.01);中、高浓度的FKQ协同ConA能升高CD4^+的比例,降低CD8^+的比例,显著升高CD4^+/CD8^+的比值(P<0.01)。此外,不同浓度的FKQ均能明显增加IFN-γ及IL-2的表达(P<0.01,P<0.05),降低IL-4和IL-10的水平,以中、高浓度最为显著(P<0.01),调节Th1/Th2平衡。结论妇科千金胶囊可有效促进脾淋巴细胞增殖,升高CD4+和CD8+比例,促进Th1细胞因子的分泌,使Th1/Th2亚群向“Th1漂移”,调节Th1/Th2平衡,提高细胞免疫应答。

Objective To study the effects of Fuke Qianjin Capsule(FKQ)on the proliferation and immunoregulation of mouse splenic lymphocytes in vitro.Methods After isolation and culture of mouse spleen lymphocytes,cells were assigned into a control group(no treatment),a ConA group(intervention with ConA),a LPS group(intervention with LPS)and Fuke Qianjin Capsule of different concentrations(FKQ).They were treated for 24 h,48 h and 72 h respectively.CCK8 assay was used to test the effects of FKQ on proliferation of spleen lymphocytes in vitro;MTT method was used to detect the effect of ConA and LPS on the proliferation of splenic lymphocytes;Flow cytometry was used to detect the proportion of CD4^+ and CD8^+ and calculate CD4^+/CD8^+ ratio.Moreover,IFN-γ,IL-2,IL-4 and IL-10 were detected by ELISA.Results FKQ at the concentrations of 30,40 and 50μg/mL significantly promoted the proliferation of splenic lymphocytes(P<0.01).FKQ could significantly promote the proliferation of T lymphocytes induced by ConA and B lymphocytes induced by LPS,with the concentration of 50μg/mL being the most significant(P<0.01).The medium and high concentrations of FKQ cooperated with ConA increased the ratio of CD4^+,decreased the ratio of CD8^+,and significantly increased the ratio of CD4^+/CD8^+(P<0.01).Different concentrations of FKQ could significantly increase the expression of IFN-γ and IL-2(P<0.01,P<0.05),decreasethe levels of IL-4 and IL-10(P<0.01),to regulated the balance of Th1/Th2.Conclusion FKQ can promote the proliferation of splenic lymphocytes effectively,increase the proportion of CD4^+ and CD8^+,and promote the secretion of Th1 cytokines,thus shifting the Th1/Th2 subpopulation to"Th1",thereby regulating the Th1/Th2 balance and improving cellular immune response.