目的研究香加皮中提取的羽扇豆烷乙酸酯(CPLA)对正常人外周血淋巴细胞和吞噬细胞免疫调节功能的影响。方法无菌分离人外周血淋巴细胞和巨噬细胞,加入PHA和不同质量浓度的CPLA,MTT法检测其对淋巴细胞增殖及巨噬细胞对食管癌细胞株ECA-109生长的影响;RT-PCR法检测CPLA对γ-干扰素(INF-γ)mRNA表达的影响;ELISA法检测CPLA作用后淋巴细胞和巨噬细胞培养上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)等细胞因子分泌的变化。结果5～40μg/mLCPLA能明显增强PHA活化的人外周血淋巴细胞的增殖(P<0.01);20、40μg/mL的CPLA能使淋巴细胞IFN-γ mRNA的表达水平明显升高(P<0.01);用CPLA处理后的巨噬细胞对肿瘤细胞的杀伤功能明显增强(P<0.01),并证实了该作用可能与CPLA促进巨噬细胞分泌TNF-α增多有关;CPLA能促进淋巴细胞产生IL-2和TNF-α,并呈现浓度依赖性(P<0.01)。结论CPLA能增强人外周血淋巴细胞和巨噬细胞的免疫功能,发挥抗肿瘤作用。

Objective Effect of Corlex Periplocae lupane acetate (CPLA) on immunoregulation of human peripheral blood lymphocytes was studied. Methods Lymphocytes from human peripheral blood were isolated and co-incubated with phytohemagglutin (PHA) and different concentration of CPLA. The effect of CPLA on the proliferation of lymphocytes and phagocytosis of macrophage to the growth of human esophageal carcinoma cells ECA-109 were measured by MTT method. Effect of CPLA on IFN-γ mRNA expression was determined by using RT-PCR. TNF-α and IL-2 secretion in supernatant of lymphocytes and macrophage culture after CPLA treatment were determined with ELISA assay. Results The proliferation of lymphocytes was enhanced by 5—40 μg/mL CPLA after activation of the proliferation of lymphocytes from human peripheral blood with PHA in a dose dependent manner (P0.01). The expression of IFN-γ mRNA in lymphocytes was enhanced by 20 and 40 μg/mL CPLA (P0.01). CPLA could strengthen the macrophage to inhibit the proliferation of esophageal carcinoma cells significantly (P0.01), which may be related with TNF-α production. The production of IL-2 and TNF-α by lymphocytes could be augmented by CPLA treatment in a dose dependent manner (P0.01). ConclusionCPLA could regulate the immune response of lymphocytes and macrophage from human peripheral blood, which may be related with killing carcinoma cells.

国家自然科学基金资助项目(30371753);河北省自然科学基金资助项目(C2004000610);河北省高校强势特色项目