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目的探讨小檗碱是否抑制脂多糖诱导的COX-2mRNA及蛋白表达,以及小檗碱是否通过ERK信号转导途径抑制COX-2的表达。方法取健康志愿者外周血,分离及培养单核细胞,分为五组,分别为对照组;脂多糖(Lipopolysaccharide,LPS)组;LPS+小檗碱25μmol/L组;LPS+小檗碱50μmol/L组;LPS+小檗碱100μmol/L组。分别在培养后30min,6h,12h,24h提取细胞,行RT-PCR法测定COX-2mRNA水平,行westernblot法测定ERK、p-ERK及COX-2蛋白水平。同时加入选择性ERK抑制剂,分别测定COX-2mRNA及蛋白水平。结果与对照组相比,LPS组COX-2mRNA及蛋白表达明显增强(P<0.01)。与LPS组相比,小檗碱组COX-2mRNA及蛋白表达明显抑制(P<0.05),且随着浓度增加,抑制作用更明显,在给药后12h,小檗碱对COX-2抑制作用最强。与LPS组相比,小檗碱组ERK活性水平有明显统计学差异(P<0.05)。加入ERK抑制剂之后,COX-2mRNA及蛋白水平降低明显(P<0.05)。结论小檗碱能抑制人外周血单核细胞COX-2mRNA及蛋白水平,其作用程度呈浓度依赖性,小檗碱对ERK活性蛋白表达有明显抑制作用。小檗碱可能通过ERK信号转导途径抑制人外周血单核细胞COX-2mRNA及蛋白表达。
Objective To investigate whether berberine can inhibit COX-2 mRNA and protein expression induced by lipopolysaccharide and whether berberine can inhibit the expression of COX-2 through ERK signal transduction pathway. Methods Peripheral blood was collected from healthy volunteers. Monocytes were isolated and cultured and divided into five groups: control group, lipopolysaccharide (LPS) group, LPS + berberine 25μmol / L group, LPS + berberine 50μmol / L Group; LPS + berberine 100μmol / L group. The cells were harvested at 30min, 6h, 12h and 24h after culture, respectively. The levels of COX-2 mRNA were determined by RT-PCR and the levels of ERK, p-ERK and COX-2 were detected by Western blotting. At the same time, selective ERK inhibitors were added to determine the COX-2 mRNA and protein levels respectively. Results Compared with the control group, the expression of COX-2 mRNA and protein in LPS group was significantly increased (P <0.01). Compared with LPS group, COX-2mRNA and protein expression of berberine group was significantly inhibited (P <0.05), and with increasing concentration, the inhibitory effect was more obvious. At 12h after administration, Berberine inhibited COX-2 The strongest Compared with LPS group, the level of ERK activity in berberine group was significantly different (P <0.05). After addition of ERK inhibitor, COX-2 mRNA and protein levels decreased significantly (P <0.05). Conclusion Berberine can inhibit the expression of COX-2 mRNA and protein in human peripheral blood mononuclear cells in a concentration-dependent manner. Berberine can significantly inhibit the expression of ERK protein. Berberine may inhibit COX-2 mRNA and protein expression in human peripheral blood mononuclear cells by ERK signal transduction pathway.