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目的:研究中药小檗碱对脂多糖(LPS)诱导的COX-2表达的影响。方法:取人外周静脉血分离及培养单个核细胞,分为3组:空白对照组、LPS组、LPS+小檗碱组。分别在培养后30 min,6 h,12 h,24 h提取细胞,行RT-PCR法测定COX-2 mRNA水平,Western blot法测定p38MAPK,p-p38MAPK,ERK,p-ERK及COX-2蛋白水平。同时加入选择性p38MAPK抑制剂(SB203580),以及特异性ERK抑制剂(PD098059)分别测定COX-2 mRNA及蛋白水平。结果:与空白对照组相比,LPS组COX-2 mRNA及蛋白表达明显增强(P<0.01)。与LPS组相比,LPS+小檗碱组COX-2 mRNA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12 h,小檗碱对COX-2抑制作用最强。与LPS组相比,LPS+小檗碱组P38MAPK活性水平差异无统计学意义(P>0.05)。但是与LPS组相比,LPS+小檗碱组ERK活性水平差异有统计学意义(P<0.05)。加入p38MAPK抑制剂以及ERK抑制剂之后,COX-2 mRNA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个核细胞COX-2 mRNA及蛋白水平,并呈浓度依赖性,但对p38MAPK活性蛋白表达无明显抑制作用;小檗碱对ERK活性蛋白表达有明显抑制作用,可能通过ERK信号转导途径抑制COX-2 mRNA及蛋白表达。
Objective: To study the effect of berberine on the expression of COX-2 induced by lipopolysaccharide (LPS). Methods: Peripheral venous blood was collected for separation and culture of mononuclear cells. The cells were divided into three groups: blank control group, LPS group and LPS + berberine group. The cells were harvested at 30 min, 6 h, 12 h and 24 h after culture, respectively. The expression of COX-2 mRNA was detected by RT-PCR and the expressions of p38 MAPK, p-p38 MAPK, ERK, p-ERK and COX- Level. At the same time, selective p38MAPK inhibitor (SB203580) and specific inhibitor of ERK (PD098059) were used to detect COX-2 mRNA and protein levels respectively. Results: Compared with the blank control group, the expression of COX-2 mRNA and protein in LPS group was significantly increased (P <0.01). Compared with LPS group, COX-2 mRNA and protein expression in LPS + berberine group was significantly inhibited (P <0.05), and with increasing concentration, the inhibitory effect was more obvious. At 12 h after administration, 2 inhibit the strongest. Compared with LPS group, the level of P38MAPK activity in LPS + berberine group had no significant difference (P> 0.05). However, compared with LPS group, the level of ERK activity in LPS + berberine group was significantly different (P <0.05). After adding p38MAPK inhibitor and ERK inhibitor, COX-2 mRNA and protein levels decreased significantly (P <0.05). CONCLUSION: Berberine can inhibit COX-2 mRNA and protein levels in human peripheral blood mononuclear cells in a concentration-dependent manner, but has no obvious inhibitory effect on p38MAPK protein expression. Berberine can significantly inhibit the expression of ERK protein , May inhibit the expression of COX-2 mRNA and protein through the ERK signal transduction pathway.