二妙散对巨噬细胞分化的调控作用研究

Effect of Ermiao Recipe on Regulating Macrophage Polarization

关键词:  二妙散  巨噬细胞  分化  炎症

Key words:  Ermiao Recipe  macrophage  polarization  inflammation

摘要:

目的 研究二妙散对脂多糖（lipopolysaccharide，LPS）诱导巨噬细胞分化的影响。方法 噻唑蓝（methyl thiazolyltetrazolium，MTT）法测定二妙散水煎液对细胞活力的影响；将LPS （100 ng·mL^-1）及不同浓度（0.1，1.0，10 mg·mL^-1）的二妙散水煎液共同作用RAW264.7细胞后，酶联免疫吸附法（enzyme-linked immune sorbent assay，ELISA）检测上清液中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白介素-10（interleukin 10，IL-10）的含量，实时荧光定量PCR （Real-timefluorescent quantitative PCR，qPCR）检测RAW264.7细胞中白介素-1β（Interleukin 1β，IL-1β），IL-6的mRNA水平，Westernblot检测RAW264.7细胞NOS2的蛋白表达情况。结果 二妙散水煎液在0.01~100 mg·mL^-1内对RAW264.7细胞无明显毒性。与LPS诱导的模型组相比，二妙散水煎液能明显降低细胞上清液中TNF-α的含量（P<0.05）、升高IL-10的含量（P<0.05），且可显著抑制RAW264.7细胞中M1分化标记物IL-1β、IL-6的mRNA水平（P<0.05）及NOS2的蛋白表达（P<0.05）。结论 二妙散可通过抑制巨噬细胞向M1促炎方向分化，从而发挥抗炎作用。

Abstract:

OBJECTIVE To investigate the effects of Ermiao Recipe on macrophage polarization induced by LPS. METHODS The cell viability was detected by MTT assay, and the contents of TNF-α and IL-10 in supernatant were determined by ELISA, the mRNA expression of IL-1β and IL-6 in RAW 264.7 were detected by qPCR and the protein expression of NOS2 in RAW 264.7 was detected by Western blot. RESULTS Ermiao Recipe water decoction significantly decreased the content of proinflammatory factor TNF-α and increased the content of anti-inflammatory factor IL-10 in cell supernatant, and also inhibited the protein expression of NOS2 and the mRNA level of IL-1β, IL-6, which were M1 polarization markers in RAW 264.7 cells. CONCLUSION Ermiao Recipe can inhibit the polarization of macrophage into proinflammatory prescription and thus exert an anti-inflammatory effect.