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目的:观察淫羊藿苷(ICA)对脂蛋白多糖诱导的巨噬细胞RAW264.7细胞M1/M2炎症表型转化作用。方法:采用M1/M2经典刺激方法后,Real-time PCR法检测M1型巨噬细胞标志基因IL-6、TNF-α和M2型巨噬细胞标志基因IL-10、Arg等的m RNA表达情况。并观察ICA不同浓度(50μg/m L、100μg/m L)以及以不同时间(6h、12h、24h)分别干预巨噬细胞M1及M2相关基因的表达变化情况。在将巨噬细胞诱导M1型后,通过Real-time PCR法检测ICA在最适浓度和最适时间干预下,M1及M2标志基因表达变化,并分析ICA对M1型巨噬细胞向M2亚型转化的诱导作用。结果:M1/M2经典方法刺激后,M1型巨噬细胞标志分子IL-6、TNF-αm RNA与M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05)。且ICA浓度为100μg/m L时,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),诱导巨噬细胞向M2方向分化。100μg/m L ICA干预12h后,对M1/M2不同标志分子m RNA表达变化产生明显影响(P<0.05)。100μg/m L ICA作用于LPS+IFN-γ诱导的M1巨噬细胞后,M2型巨噬细胞标志分子IL-10、Arg m RNA表达明显升高(P<0.05),M1型巨噬细胞标志分子IL-6 m RNA表达下降(P<0.05)。结论:ICA可以诱导体外培养巨噬细胞RAW264.7向抗炎亚型M型方向分化,并可以诱导已经分化的促炎亚型M1巨噬细胞向抗炎亚型M2方向转化。
Objective: To observe the effect of icariin (ICA) on lipopolysaccharide-induced M1 / M2 inflammatory phenotype transformation in RAW264.7 macrophages. Methods: After M1 / M2 classic stimulation, Real-time PCR was used to detect the mRNA expression of IL-6, TNF-α and M2 markers of macrophage marker type M1 . The expressions of M1 and M2 related genes in macrophages were observed under different concentrations of ICA (50μg / ml, 100μg / ml) and at different time (6h, 12h, 24h) respectively. After induced by macrophages, the expression of M1 and M2 markers in ICA were detected by Real-time PCR and the effects of ICA on M1 subtype to M2 subtype Induction of transformation. Results: After M1 / M2 stimulation, the expressions of IL-6, TNF-αmRNA and IL-10 and Arg m RNA of M1 macrophage markers were significantly increased (P <0.05). When the ICA concentration was 100 μg / mL, the expression of IL-10 and Arg m RNA of M2 macrophages was significantly increased (P <0.05), which induced the differentiation of macrophages to M2. After 12 h of ICA treatment at 100 μg / mL, the expression of m RNA in different molecular markers of M1 / M2 significantly changed (P <0.05). The expression of IL-10 and Arg m RNA of M2 macrophages was significantly increased after treated with 100 μg / mL ICA for LPS + IFN-γ-induced M1 macrophages (P <0.05) IL-6 mRNA expression decreased (P <0.05). CONCLUSION: ICA can induce macrophage RAW264.7 to differentiate into anti-inflammatory subtype M in vitro and induce the differentiation of already differentiated pro-inflammatory subtype M1 macrophages to anti-inflammatory subtype M2.