二氢杨梅素对H2O2诱导PC12细胞损伤的保护作用研究

Protective Effect of Dihydromyricetin on PC12 Cell Injury Induced by H2O2

目的研究二氢杨梅素(dihydromyricetin,DHM)对H 2O 2诱导的PC12细胞凋亡的影响及机制。方法建立H 2O 2诱导的PC12细胞氧化损伤模型,以不同浓度二氢杨梅素(10、20、40μmol/L)处理细胞,采用噻唑蓝(MTT)比色法检测二氢杨梅素对PC12细胞活性的影响,ELISA法检测细胞上清液乳酸脱氢酶的活性,Hoechst 33342染色检测细胞凋亡,免疫印迹法检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和天冬氨酸蛋白水解酶(caspase-3)的表达情况。结果MTT和LDH结果显示H 2O 2100μmol/L显著损伤PC12细胞,二氢杨梅素(10、20、40μmol/L)可保护H 2O 2诱导的细胞损伤。Hoechst染色结果显示,与模型组比较,二氢杨梅素(10、20、40μmol/L)显著抑制细胞的凋亡率(P<0.01)。Western blot法结果显示,与模型组比较,二氢杨梅素显著抑制H 2O 2诱导的PC12细胞Bax和caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论二氢杨梅素通过抑制细胞凋亡保护H 2O 2诱导的PC12细胞氧化损伤。

Objective To investigate the effect and mechanism of dihydromyricetin(DHM)on cell apoptosis of PC12 cells induced by H 2O 2.Methods PC12 cells are treated with H 2O 2 to induce cell injury,administrated with different concentrations of DHM(10,20,40μmol/L).PC12 cell viability was evaluated by MTT detection assay,and the activity of lactate dehydrogenase(LDH)in cell supernatant was detected by ELISA detection assay.Hoechst 33342 staining was used to detect cell apoptosis.The expression of Bcl-2,Bcl-2-related X protein(Bax)and aspartate proteolytic enzyme(caspase-3)were detected by Western blot.Results MTT and LDH results showed that H 2O 2100μmol/L significantly damaged PC12 cells,and DHM(10,20,40μmol/L)could protect the PC12 cells from H 2O 2-induced cell injury.Hoechst 33342 staining showed that DHM(10,20,40μmol/L)significantly decreased the apoptotic rate of PC12 cells compared with model group(P<0.01).Western blot results showed that DHM significantly inhibited the up-regulation of Bax and caspase-3 protein expression and the down-regulation of Bcl-2 protein expression in PC12 cells compared with model group.Conclusion Dihydromyricetin can protect PC12 cells against cellular damage induced by H 2O 2 through inhibiting cell apoptosis.