论文部分内容阅读
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 investigate the possible mechanism of Sanhuang Yinchi Decoction in treating acute liver injury. Methods Human embryonic liver cell line LO2 was divided into three groups: normal group, model group, control group and Sanhuang Yinchuang high, medium and low dose groups, D-Gal N induced LO2 cell necrosis, The Huangyinchitang high, medium and low dose groups were given Sanhuang Yinchuan 3,2,1 mg / ml, the control group was given receptor interacting protein 1 inhibitor (Nec-1) 60 mmol / L, MTT Method was used to determine the cell viability in each group. Hoechst and PI staining was used to detect the apoptosis and necrosis of the cells. The apoptosis rate and intracellular reactive oxygen species (ROS) levels were detected by flow cytometry. The expressions of Casepase-3 , Receptor interacting protein 1 (RIP1) and receptor interacting protein 3 (RIP3). Results Compared with the model group, Sanhuang Yinchu medium and low dose groups could improve cell survival rate and ROS clearance rate, Sanhuang Yinchuan medium dose group could reduce the fluorescence intensity of Hoechst and PI staining, High-dose group also increased ROS clearance (P <0.05). Sanhuang Yin Chuan decoction medium dose group to reduce apoptosis rate of late cells to improve the survival rate of the best. Compared with the Sanhuang Yinchu decoction group, the effect of the control group was more obvious (P <0.01). Conclusion Sanhuang Yinchuang can protect D-Gal N-induced LO2 cell death by inhibiting the expression of RIP1 and RIP3.