甘草总黄酮对阿司匹林损伤人胃黏膜上皮细胞的保护作用及机制研究

Mechanism of Licoflavone in Protecting Human Gastric Mucosal Epithelial Cells from Aspirin-induced Injury

目的探讨甘草总黄酮对阿司匹林损伤人胃黏膜上皮细胞(GES-1)的保护作用及可能机制。方法 2017年3—11月,选取GES-1细胞株,分为6组,对照组:加DMEM培养基;阿司匹林组:阿司匹林18.5 mmol/L与GES-1共同培养24 h;甘草总黄酮低组、甘草总黄酮中组、甘草总黄酮高组:GES-1分别以甘草总黄酮(3、6、12mg/L)预处理2 h后再加阿司匹林18.5 mmol/L共同培养24 h;PD98059组:GES-1以甘草总黄酮12 mg/L预处理2 h后,加PD98059 10μmol/L处理1 h,再加阿司匹林18.5 mmol/L共同培养24 h。采用MTT法检测细胞增殖抑制率,采用流式细胞术检测细胞凋亡率,检测乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量,采用Western blotting法检测细胞磷酸化细胞外调节蛋白激酶(p-ERK1/2)、B淋巴细胞癌2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达。结果与对照组比较,阿司匹林组、甘草总黄酮低组、甘草总黄酮中组、甘草总黄酮高组、PD98059组细胞增殖抑制率、细胞凋亡率、LDH活性、MDA含量、p-ERK1/2、Bax表达升高,SOD活性、Bcl-2表达降低(P<0.05);与阿司匹林组比较,甘草总黄酮中组、甘草总黄酮高组、PD98059组细胞增殖抑制率、细胞凋亡率、LDH活性、MDA含量、p-ERK1/2、Bax表达降低,SOD活性、Bcl-2表达升高(P<0.05);与甘草总黄酮低组、甘草总黄酮中组、甘草总黄酮高组比较,PD98059组细胞增殖抑制率、细胞凋亡率、LDH活性、MDA含量、p-ERK1/2、Bax表达降低,SOD活性、Bcl-2表达升高(P<0.05)。结论甘草总黄酮可通过促增殖、抗凋亡、抗氧化应激等途径减轻阿司匹林诱导的GES-1损伤,其机制可能与抑制ERK1/2信号通路有关。

Objective To investigate the effect of licoflavone on aspirin-induced human gastric mucosal epithelial(GES-1)cell injury and its mechanism.Methods The study was conducted from March to November 2017.GES-1 cells were divided into 6 groups,control group(incubated with DMEM),aspirin group(incubated with 18.5 mmol/L aspirin for 24 h),low-dose,medium-dose and high-dose licoflavone groups(incubated with 18.5 mmol/L aspirin for 24 h after being pre-treated with 3,6,12 mg/L licoflavone for 2 h,respectively),PD98059 group(incubated with 18.5 mmol/L aspirin for 24 h after being pre-treated with 12 mg/L licoflavone for 2 h and with 10μmol/L PD98059 for 1 h successively).Proliferation inhibition rate was detected by MTT assay,apoptosis rate was measured by flow cytometry,lactate dehydrogenase(LDH),superoxide dismutase(SOD)activities,and malondialdehyde(MDA)content were determined by colorimetry,the protein expressions of p-ERK1/2,Bcl-2 and Bax were measured by western blotting.Results The control group showed significantly decreased proliferation inhibition rate,apoptosis rate,LDH activity,MDA content,and p-ERK1/2 and Bax expressions,but obviously increased SOD activity and Bcl-2 expression compared with other groups,as did the medium-dose and high-dose licoflavone groups and PD98059 group when compared with the aspirin group,and the PD98059 group when compared with the 3 licoflavone groups(P<0.05).Conclusion Licoflavone alleviates aspirin-induced GES-1 cell injury through pro-proliferation,antiapoptosis and anti-oxidative stress via the inhibition of ERK1/2 signaling pathway.