CeNPs对DOX诱导的心肌损伤的作用及其对PGC1-α的影响

Effect of CeNPs on DOX-induced Myocardial Injury and its Effect on PGC1-α

目的探讨CeNPs对DOX诱导的心肌损伤的作用及具体机制。方法培养H9C2心肌细胞,将生长良好的H9C2心肌细胞随机分为5组:正常组、DOX组、CeNPs组、CeNPs+DOX组、DEX+DOX组,倒置显微镜下观察细胞形态。CCK-8实验测定各组的OD值,检测细胞活力;流式细胞术检测细胞凋亡率;Western Blot检测PGC1-α及凋亡相关蛋白Bax、Bcl-2表达。结果(1)细胞形态学:DOX处理后的细胞大量凋亡,CeNPs+DOX处理后的H9C2心肌细胞可使凋亡程度减轻。(2)CCK-8法显示:①DOX组较正常组细胞活力明显下降;②DOX与不同浓度CeNPs组相比,CeNPs为10μg/mL时,OD值最高,细胞活力增高;③以浓度为10μg/mL CeNPs为最适保护心肌细胞用药浓度,后续实验均使用该浓度;④CeNPs及DEX处理后可显著增加H9C2心肌细胞活力。(3)流式细胞术显示:与正常组相比,DOX组的细胞凋亡率升高;与DOX组相比,CeNPs+DOX组和DEX+DOX组的细胞凋亡率降低且差异具有统计学意义(P<0.0001)。(4)Western Blot结果:①与正常组相比,DOX组Bax表达量显著升高,Bcl-2、PGC1-α表达显著降低,且Bcl-2/Bax比值两两比较后差异均有统计学意义(P<0.05);②与DOX组相比,CeNPs+DOX组和DEX+DOX组的Bax表达量降低,Bcl-2、PGC1-α的表达量升高,Bcl-2/Bax比值升高,CeNPs+DOX组和DEX+DOX组的Bax、Bcl-2、PGC1-α的表达水平相比较,差异无统计学意义(P>0.05)。结论(1)CeNPs通过抑制凋亡保护DOX诱导的心肌细胞损伤。(2)PGC1-α在CeNPs保护DOX心肌细胞损伤中高表达。

Objective To explore the role and specific mechanism of CeNPs in DOX-induced myocardial injury.Methods H9C2 cardiomyocytes were cultured and randomly divided into five groups:normal group,DOX group,CeNPs group,CeNPs+DOX group and DEX+DOX group.The cell morphology was observed under the in⁃verted microscope.CCK-8 experiment was used to determine the OD value and cell viability of each group.Apop⁃tosis rate was detected by flow cytometry.Western-Blot method was used to detect the expression of PGC1-αand apoptosis-related proteins Bax and Bcl-2.Results(1)Cell morphology:A large number of cells were apoptotic after DOX treatment,and H9C2 cardiomyocytes treated with CeNPs+DOX could reduce the degree of apoptosis.(2)CCK-8 method showed that:①compared with the normal group,the cell viability of DOX group decreased significantly;Compared with different concentrations of CeNPs;DOX had the highest OD value and increased cell vi⁃ability when CeNPs was 10 ug/mL.③The concentration of 10ug/mLCeNPs was the optimal concentration for pro⁃tecting myocardial cells,which was used in subsequent experiments.④CeNPs and DEX can significantly increase the viability of H9C2 cardiomyocytes.(3)Flow cytometry showed that the apoptosis rate of DOX group was higher than that of normal group.Compared with DOX group,the apoptosis rate of CeNPs+DOX group and DEX+DOX group decreased with statistical significance.(4)Western Blot results:①Compared with the normal group,the ex⁃pression of Bax in DOX group was significantly increased,while the expressions of Bcl-2 and PGC1-αwere sig⁃nificantly decreased,and the ratio of Bcl-2/Bax was statistically significant;②Compared with DOX group,the ex⁃pression of Bax in CeNPs+DOX group and DEX+DOX group decreased,the expression of Bcl-2 and PGC1-αincreased,and the ratio of Bcl-2/Bax increased.Conclusion(1)CeNPs protects cardiomyocytes from DOX-in⁃duced injury by inhibiting apoptosis.(2)PGC1-αis highly expressed in the myocardial cell injury of DOX protec⁃ted by CeNPs.