绿原酸对小鼠脾脏淋巴细胞体外增殖和分泌功能的影响

为分析绿原酸(CHA)对小鼠脾脏淋巴细胞增殖、分泌功能的影响。无菌操作分离小鼠脾脏，制备脾脏淋巴细胞并用含10%胎牛血清的RPMI 1640培养，在培养液中分别加入诱导剂刀豆蛋白(ConA)和脂多糖(LPS)以及0、40、80、160 μg/mL不同浓度的CHA，经过不同时间培养后，采用MTT法检测T和B淋巴细胞的增殖，ELISA和Griess法分别检测淋巴细胞分泌TNF-α和NO的水平。结果表明：低浓度和中浓度CHA提高ConA/LPS诱导的T、B淋巴细胞增殖和存活，而高浓度CHA明显抑制其细胞增殖转化；CHA促进小鼠脾脏T、B淋巴细胞分泌NO；CHA对T淋巴细胞分泌TNF-α影响不大，中浓度CHA可显著促进脾脏B淋巴细胞分泌TNF-α，高浓度反而抑制其分泌；CHA促进环磷酰胺处理的小鼠脾脏T和B淋巴细胞体外增殖，但对其NO分泌量影响不明显。结果提示CHA可能通过影响淋巴细胞增殖和细胞因子分泌而表现免疫调节作用。

Effects of Chlorogenic Acid on Proliferations and Secretion Function of Mouse Spleen Lymphocytes in vitro

To investigate effects of Chlorgenic Acid（CHA） on proliferations and secretion function of mouse spleen lymphocytes in vitro.Mouse spleen cells were prepared by aseptic separation and in vitro cultured in RPMI 1640 containing 10% fetal blood serum.The stimulant Concanavalin（ConA） and Lipopolysaccharide（LPS） were added in culture medium with different CHA concentrations of 40,80,160 μg/mL.After cells were cultured for different time,MTT assay the proliferations of T and B lymphocytes,ELISA and Griess methods were used to detected TNF-α and NO levels.The results showed that proliferation and survival rate of ConA-induced T lymphocytes and LPS-induced B lymphocytes treated with low and medium CHA concentration were improved,and cellular transformation significantly inhibited at high concentration of CHA.CHA could enhance the NO secretion of T and B lymphocytes treated with CHA;TNF-α secretion of T lymphocytes basically did not be affected.TNF-α secreted from spleen B lymphocytes was promoted at medium concentration of CHA,but inhibited at high CHA concentration.CHA promoted the proliferation of spleen T and B lymphocytes from mice treated with cyclophosphamide,but there was not a significantly effect of CHA on NO secretion.The results suggested that CHA might affect lymphocyte proliferation and cytokine secretion to perform immunomodulatory effects.