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该文主要研究4种黄连生物碱对小鼠腹腔巨噬细胞的影响,初步探讨其调节机制。MTT法检测黄连生物碱对巨噬细胞活力的影响;采用中性红实验和NBT法分别测定生物碱对巨噬细胞吞噬和呼吸爆发活性的影响;通过实时荧光定量PCR检测黄连生物碱处理后巨噬细胞呼吸爆发相关基因的表达量;用荧光分光光度计检测生物碱对巨噬细胞膜蛋白构象的影响。结果显示,4种黄连生物碱对小鼠腹腔巨噬细胞的吞噬功能都有不同程度的提高,小檗碱效果最好;小檗碱、黄连碱和巴马汀对佛波酯(PMA)刺激的小鼠腹腔巨噬细胞呼吸爆发活性具有正向调节的作用,并且能够在一定程度上增加巨噬细胞中PKC,p40phox或p47phox mRNA的表达量,而表小檗碱对呼吸爆发没有显著性影响;黄连生物碱能够改变巨噬细胞膜蛋白的构象,其中小檗碱作用最强。结果表明黄连生物碱可能通过改变巨噬细胞膜蛋白构象从而激活巨噬细胞,增强其吞噬和呼吸爆发功能,而且,生物碱对巨噬细胞呼吸爆发的调节机制还可能与相关基因PKC,p40phox和p47phox mRNA的表达有关。
This paper mainly studies the effects of four kinds of Coptis alkaloids on murine peritoneal macrophages and initially explores the regulatory mechanism. The effect of Coptis alkaloids on the viability of macrophages was detected by MTT assay. The effect of alkaloids on macrophage phagocytosis and respiratory burst activity was determined by neutral red assay and NBT assay. The expression of genes related to respiratory burst in macrophages was measured. The effect of alkaloids on the conformation of membrane proteins of macrophages was detected by fluorescence spectrophotometer. The results showed that the four kinds of Coptis alkaloids on phagocytosis of mouse peritoneal macrophages have varying degrees of improvement, the best berberine; berberine, Coptis and palmatine stimulation of phorbol ester (PMA) Of mouse peritoneal macrophages respiratory burst activity has a positive regulatory role, and to a certain extent, increase the expression of macrophages in PKC, p40phox or p47phox mRNA, and epiberberine on respiratory burst did not have a significant effect Coptis alkaloids can change the conformation of macrophage membrane protein, of which berberine has the strongest effect. The results showed that Coptis alkaloids may activate the macrophages by altering the membrane protein conformation of macrophages and enhance their phagocytosis and respiratory burst function. Moreover, the regulatory mechanism of alkaloids on respiratory burst of macrophages may be related to the related genes PKC, p40phox and p47phox mRNA expression.