蓟黄素抗炎和免疫调节作用的研究

Study on the anti-inflammatory and immunomodulatory effects of cirsimaritin

目的 研究蓟黄素的抗炎及和免疫调节作用。方法 采用环氧合酶(COX 1/2)和5-脂氧酶(5-LO)抑制剂筛选试剂盒检测蓟黄素对COX 1/2和5-LO的抑制作用。采用细菌脂多糖(LPS)刺激小鼠巨噬细胞RAW 264.7细胞建立炎症细胞模型,经蓟黄素处理后,采用Griess试剂检测一氧化氮(NO)的含量;2′,7′-氯荧光黄双乙酸酯荧光探针法检测细胞内活性氧(ROS)的释放情况;酶联免疫法(ELISA)检测促炎症因子的水平;RT-qPCR法测定炎症因子mRNA的表达水平。采用MTS法检测蓟黄素对小鼠静息期脾淋巴细胞、刀豆蛋白A(ConA)诱导的T淋巴细胞和LPS诱导的B淋巴细胞增殖的影响;ELISA法检测蓟黄素对T淋巴细胞诱生细胞因子的影响。结果 蓟黄素对COX-1和COX-2的半数抑制浓度分别为192.06、57.21μg·mL,蓟黄素对5-LO的活性无明显影响。蓟黄素能明显抑制LPS诱导RAW264.7细胞中NO、ROS、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、单核细胞趋化蛋白-1和前列腺素E的生成,明显下调TNF-α、IL-6和一氧化氮合酶的mRNA的表达。蓟黄素可明显减少ConA/LPS诱导的T/B淋巴细胞增殖,并明显降低T淋巴细胞分泌的细胞因子。结论 蓟黄素具有良好的抗炎活性,并能抑制ConA/LPS诱导的T/B淋巴细胞的增殖及细胞因子的分泌。

OBJECTIVE To investigate anti-inflammatory and immunomodulatory effects of cirsimaritin.METHODS Cyclooxygenase 1/2(COX 1/2) and 5-lipoxygenase(5-LO) inhibitor screening were employed to evaluate the inhibitory effects of cirsimaritin on COX 1/2 and 5-LO.RAW264.7 cells were pretreated with cirsimaritin, and then the inflammatory models established by stimulating with lipopolysaccharide(LPS).NO level was assessed using Griess reagent.The level of intracellular ROS was detected by 2′,7′-dichlorodihydrofluorescein diacetate fluorescence probe.The contents of inflammatory cytokines were detected by ELISA.The expression of inflammatory cytokines mRNA was determined by RT-qPCR.The proliferative effects of cirsimaritin on mouses spleen lymphocytes, T and B lymphocytes induced by conA and LPS were tested by MTS.The effects of cirsimaritin on secretion of cytokines in T lymphocytes were assayed by ELISA.RESULTS The ICvalues of cirsimaritin to COX-1 and COX-2 were 192.06 and 57.21 μg·mL,respectively.Cirsimaritin had no effect on 5-LO.Cirsimaritin significantly inhibited LPS-induced NO,ROS,TNF-α,IL-6,MCP-1 and PGEcytokines production in RAW264.7 cells, and inhibited the mRNA expression of TNF-α,IL-6 and iNOS.Cirsimaritin could reduce T and B lymphocytes proliferation and the secretion of cytokines in T lymphocytes.CONCLUSION Cirsimaritin possesses excellent anti-inflammatory activity.Cirsimaritin can inhibit T and B lymphocytes proliferation and the secretion of cytokines.