小檗碱减轻幽门螺杆菌诱导的人胃黏膜上皮细胞损伤

目的:探讨小檗碱(Ber)对幽门螺杆菌(Hp)诱导的人胃黏膜上皮细胞GES-1损伤的保护作用及机制。方法:采用Hp感染GES-1细胞,加入小檗碱(5,10和20μmol/L)和细胞外调节蛋白激酶(ERK1/2)抑制剂PD98059(20μmol/L)共培养。MTT法检测细胞活力,流式细胞术检测细胞凋亡,ELISA法检测细胞白细胞介素(IL)-1β及IL-8的水平,比色法检测细胞乳酸脱氢酶(LDH)活性并以Western blot法检测细胞Bax、Bcl-2、p-ERK1/2的蛋白水平。结果:与对照组比较,Hp可抑制GES-1细胞活力、促进GES-1细胞凋亡、诱导GES-1细胞分泌IL-1β和IL-8、增加LDH活性、增加GES-1细胞内p-ERK1/2及Bax蛋白水平并降低Bcl-2蛋白水平,差异均有统计学显著性(P0.05);与Hp感染组比较,中、高剂量小檗碱均可逆转Hp对GES-1细胞活力、细胞凋亡、细胞IL-1β及IL-8分泌、LDH活性和GES-1细胞内p-ERK1/2、Bax及Bcl-2蛋白水平的影响,差异均有统计学显著性(P0.05);与高剂量小檗碱组比较,PD98059联合小檗碱可进一步逆转Hp对GES-1细胞上述指标的影响,差异均有统计学显著性(P0.05)。结论:小檗碱可通过抗炎、增加细胞活力和抗凋亡来减轻Hp诱导的GES-1细胞损伤,其机制可能与抑制ERK1/2通路有关。

Effect of berberine on Helicobacter pylori-induced human gastric epithe-lial cells injury

AIM: To investigate the effect of berberine (Ber) on Helicobacter pylori (Hp)-induced human gastric epithelial cells (GES-1) injury and the underlying mechanism. METHODS: Berberine (5, 10 and 20 μmol/L) and PD98059 (20 μmol/L), a selective inhibitor of extracellular regulated protein kinases (ERK)1/2 signaling pathway, were added to Hp-infected GES-1 cells. The cell activity and apoptosis, the levels of interleukin (IL)-1β and IL-8, lactic dehydrogenase (LDH) activity and the protein levels of Bax, Bcl-2 and p-ERK1/2 in the GES-1 cells were determined by MTT assay, flow cytometry, ELISA, colorimetry and Western blot, respectively. RESULTS: Compared with control group, Hp significantly inhibited the cell activity, increased the apoptotic rate, LDH activity, IL-1β and IL-8 levels, the Bax and p-ERK1/2 protein levels but decreased the Bcl-2 protein level in GES-1 cells (P<0.05). However, these effects of Hp were reversed by berberine at medium-dose and high-dose, as compared with the Hp-infected GES-1 cells (P<0.05). Moreover, the protective effects of berberine were significantly enhanced by the co-incubation of berberine with PD98059, as compared with the berberine at higher dose (P<0.05). CONCLUSION: Berberine may attenuate Hp-induced human gastric epithelial GES-1 cells injury by anti-inflammation, promoting cell growth and anti-apoptosis via the inhibition of ERK1/2 signaling pathway.