三黄茵赤汤对D-氨基半乳糖诱导人胚胎肝细胞系LO2坏死的影响  

Impact of Sanhuang Yinchi Decoction (三黄茵赤汤) on Human Embryonic Liver Cell Line LO2 Necrosis Induced by D-galactosamine

作　　者：李海叶[1] 刁建新[1] 代欢[1] 黄伟[1] 杨运高[1] 

机构地区：[1]南方医科大学中医药学院

出　　处：《中医杂志》2017年第9期782-786,共5页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81673820;81603606)

摘　　要：目的探讨三黄茵赤汤治疗急性肝损伤的可能作用机制。方法将人胚胎肝细胞系LO2分为正常组、模型组、对照组及三黄茵赤汤高、中、低剂量组,通过D-氨基半乳糖(D-Gal N)诱导LO2细胞坏死,三黄茵赤汤高、中、低剂量组分别给予三黄茵赤汤3、2、1 mg/ml,对照组给予受体相互作用蛋白1抑制剂(Nec-1)60 mmol/L,采用MTT法测定各组细胞存活率,Hoechst、PI染色分析细胞是否发生凋亡、坏死,流式细胞仪检测细胞凋亡率及细胞内活性氧(ROS)水平,Western blot法检测各组细胞Casepase-3、受体相互作用蛋白1(RIP1)、受体相互作用蛋白3(RIP3)表达情况。结果与模型组比较,三黄茵赤汤中、低剂量组均能提高细胞生存率和ROS的清除率,三黄茵赤汤中剂量组能降低Hoechst、PI染色荧光强度,三黄茵赤汤高剂量组亦能提高ROS的清除率(P<0.05)。三黄茵赤汤中剂量组降低细胞晚期凋亡率、提高生存率的效果最好。三黄茵赤汤各剂量组均能减少RIP1及RIP3表达(P<0.05);与三黄茵赤汤各剂量组相比,对照组效果更加明显(P<0.01)。结论三黄茵赤汤通过抑制RIP1及RIP3的表达而对D-Gal N诱导LO2细胞坏死产生保护作用。Objective To explore the possible mechanism of Sanhuang Yinchi Decoction（ 三黄茵赤汤） in treating acute liver injury. Methods Human embryonic liver cell line LO2 were divided into normal group,model group,control group and high,medium,and low dose Sanhuang Yinchi Decoction groups. D-galactosamine（ DGal N） was used to induce LO2 cells necrosis. The high,medium,and low dose Sanhuang Yinchi Decoction groups were given Sanhuang Yinchi Decoction 3,2,1 mg/ml respectively. The control group was given receptor interacting protein 1 inhibitor（ Nec-1） 60 mmol/L. The survival rate of cells in each group was determined by MTT method.Apoptosis and necrosis were analyzed by Hoechst and propidium iodide（ PI） staining. The apoptosis rate and the level of intracellular reactive oxygen species（ ROS） were detected by flow cytometry. The expressions of Casepase-3,receptor interacting protein1（ RIP1） and receptor interacting protein3（ RIP3） were detected by Western blot method.Results Compared with the model group,the survival rate of cells and ROS clearance in the medium and low dose Sanhuang Yinchi Decoction groups increased. The fluorescence intensity of Hoechst and PI staining in the mediumdose Sanhuang Yinchi Decoction group decreased. The ROS clearance in the high-dose Sanhuang Yinchi Decoction group increased（ P〈0. 05）. The effect of decreasing the late cell apoptosis rate and increasing the survival rate in the medium-dose Sanhuang Yinchi Decoction group was the best. The expressions of RIP1 and RIP3 in all dose Sanhuang Yinchi Decoction groups decreased（ P〈0. 05）. Compared with all dose Sanhuang Yinchi Decoction groups,the effect in the control group was more obvious（ P〈0. 01）. Conclusion Sanhuang Yinchi Decoction might have protection on LO2 cells necrosis induced by D-Gal N by inhibiting the expressions of RIP1 and RIP3.

关 键 词：急性肝损伤 三黄茵赤汤 D-氨基半乳糖 细胞坏死 受体相互作用蛋白1 受体相互作用蛋白3 

分 类 号：R285[医药卫生—中药学]