连翘酯苷对小鼠脾脏淋巴细胞体外增殖与分泌功能的影响

摘要

Objective To investigate the effects of forsythoside (FS) on immune regulatory mechanism of proliferation and secretion of NO and TNF-ot of mouse spleen T and B lymphocytes in vitro. Methods Mouse spleen and spleno-cytes were prepared by aseptic separation and the cells were suspended with RPMI 1640 and cultured in RPM1 1640 containing 10% fetal bovine serum in vitro and stimulant concanavalin ( Con A ) and lipopolysaccharide ( LPS) . Different concentrations of 40, 80 and 160 μg/mL FS were added separately in culture medium at different times. MTT was used to assay the changes of optical absorbance of T and B lymphocytes. ELISA and Criess methods were used to detect the TNF-α and NO levels. Results Proliferation and survival rate of ConA-induced T lymphocytes treated with FS at low and medium concentration were significantly increased at 24 h and 48 h, and cell transformation was inhibited after FS induction for 72 h. LPS-induced splenic B cell proliferation and survival were significantly improved by FS at low concentration after 24 h. NO secretion of T and B lymphocytes was enhanced by FS at low and medium concentrations. TNF-α secretion of FS-treated B and T lymphocytes treated with FS at medium concentration was promoted, but TNF-α secretion of T lymphocytes was inhibited by FS at high concentration. In addition, FS had a significant effect on proliferation of splenic lymphocytes in vitro from mouse injected with cyclophosphamide, but no significant effect on the lymphocyte NO secretion. Conclusion Our findings suggest that FS may regulate immune cell function via lymphocyte proliferation and cytokine secretion.%目的 分析连翘酯苷(FS)对小鼠脾脏T和B淋巴细胞增殖、分泌NO和TNF-α的影响,初步探讨其免疫调节作用机制.方法 无菌操作分离小鼠脾脏,制备脾脏细胞并用含10％胎牛血清的RPMI 1640培养,在培养液中分别加入刺激刺刀豆蛋白( ConA)和脂多糖(LPS)以及不同浓度40、80、160 μg/mL的FS共培养不同时间,采用MTT法检测T和B淋巴细胞的吸光度变化,ELISA和Griess法分别检测细胞分泌TNF-α和NO的水平.结果低浓度和中浓度FS对ConA诱导T淋巴细胞24 h和48 h后细胞增殖和存活率明显提高,诱导时间延长至72 h后FS明显抑制细胞转化;低浓度FS对LPS诱导脾脏B淋巴细胞24 h后细胞增殖和生存率显著提高;FS促进小鼠脾脏T和B淋巴细胞分泌NO;FS促进B淋巴细胞分泌TNF-α,中浓度FS促进T淋巴细胞分泌TNF-α而高浓度反而抑制其分泌.此外,FS对环磷酰胺(CY)处理小鼠的脾脏淋巴细胞体外增殖有明显影响,对细胞NO分泌影响不显著.结论 结果提示FS可能通过影响小淋巴细胞增殖和细胞因子分泌而调节免疫细胞功能.