朝天罐总酚对脂多糖诱导的小鼠巨噬细胞炎症反应的调控作用

Regulatory effect of Osbeckia opipara total phenols on lipopolysaccharide-induced inflammatory response in mouse macrophages

目的:探讨朝天罐总酚对脂多糖(LPS)诱导小鼠巨噬细胞(RAW264.7)炎症反应的调控作用。方法:培养小鼠RAW264.7巨噬细胞,以LPS诱导构建细胞炎症模型,采用MTT法测定不同浓度(1.88μg/mL、3.75μg/mL、7.5μg/mL、15μg/mL、30μg/mL、60μg/mL)朝天罐总酚对细胞增殖的影响,中性红吞噬实验检测朝天罐总酚对RAW264.7细胞吞噬能力的影响。根据细胞存活率和吞噬功能检测结果,设为朝天罐总酚低、中、高剂量组,Griess法检测细胞上清液一氧化氮(NO)含量,酶联免疫吸附试验(ELISA)法检测细胞上清液中肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6水平。结果:朝天罐总酚浓度≤15μg/mL时,其对RAW264.7细胞存活率无显著影响(P>0.05)。与空白对照比较,3.75~15μg/mL朝天罐总酚可增强RAW264.7细胞细胞吞噬能力(P<0.01)。与空白对照组比较,模型组NO、TNF-α、IL-6水平升高(P<0.01);与模型组比较,朝天罐总酚中、高剂量组NO、TNF-α水平降低,朝天罐总酚各剂量组IL-6水平降低(均P<0.01)。结论:朝天罐总酚可能通过抑制炎症因子产生、增强细胞吞噬功能对LPS诱导的RAW264.7巨噬细胞发挥抗炎作用。

Objective:To investigate the regulatory effect of total phenols from Osbeckia opipara on lipopolysaccharide(LPS)-induced inflammatory response in mouse macrophages RAW264.7.Methods:Mouse RAW264.7macrophages were cultured,and a cellular inflammation model was constructed with LPS induction.The effects of different concentrations(1.88μg/mL,3.75μg/mL,7.5μg/mL,15μg/mL,30μg/mL,60μg/mL)of Osbeckia opipara total phenols on cell proliferation were determined by MTT assay,and the neutral red phagocytosis assay was performed to detect the effects of Osbeckia opipara total phenols on the phagocytic ability of RAW264.7cells.According to the results of cell viability and phagocytosis assay,low,medium and high dose groups were set up.The level of nitric oxide(NO)in cell supernatant was detected by Griess method.The tumor necrosis factor(TNF)-αand interleukin(IL)-6 in cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Results:Osbeckia opipara total phenols had no significant effect on the survival rate of RAW264.7cells at concentrations≤15μg/mL(P>0.05).Compared with the blank control,3.75-15μg/mL of Osbeckia opipara total phenols enhanced the phagocytic ability of RAW264.7 cells(P<0.01).Compared with the blank control group,NO,TNF-αand IL-6 levels were increased in the model group(P<0.01).Compared with the model group,NO and TNF-αlevels were decreased in the middle and high dose groups of Osbeckia opipara total phenols,and IL-6 level was decreased in each dose group(P<0.01).Conclusion:Osbeckia opipara total phenols may exert anti-inflammatory effects on LPS-induced RAW264.7 macrophages by inhibiting inflammatory factors production and enhancing the phagocytosis.