银杏叶提取物EGB761对D-半乳糖诱导的心肌细胞老化的保护作用

Evaluation of prevention effects of Extract of Ginkgo Biloba(EGB761)on aging cardiomyocytes cell induced by D-galactose

目的:探讨银杏叶提取物EGB761对D-半乳糖诱导的心肌细胞老化的影响及其可能机制。方法:用5 g/L D-半乳糖处理培养心肌细胞建立衰老模型,并以不同浓度EGB761(5、10、20μg/mL)分别干预48 h。以β-半乳糖苷酶染色判断细胞衰老,ELISA法检测心肌细胞晚期糖基化终产物(advanced glycation end products,AGEs)的含量。显微镜下观察心肌细胞的形态和搏动情况。结果:与正常对照组相比,D-半乳糖诱导的心肌细胞内β-半乳糖苷酶染色阳性率显著提高(75.6%±4.9%vs 17.2%±2.9%,P<0.01);细胞内AGEs含量明显增高[(703±32)vs(93±26)pg/mL,P<0.01];同时出现细胞形态学和搏动的异常。与D-半乳糖组比较,不同浓度(5、10、20μg/mL)EGB761干预组β-半乳糖苷酶染色阳性率显著下降(各组分别为57.7%±7.9%、49.7%±9.2%、34.0%±6.6%,P<0.01),同时AGEs含量显著降低[分别为(404±33)、(357±25)、(249±77)pg/mL,P<0.01],心肌细胞的搏动明显增加。结论:EGB761可通过抑制心肌细胞的非酶糖基化,预防D-半乳糖诱导的心肌细胞老化。

AIM：To investigate the preventive effects of Extract of Ginkgo Biloba （EGB761） on the cell aging of cardiomyocytes induced by D-galactose and its latent mechanism. METHODS：Cell aging of cultured neonatal rat cardiomyocytes was created by 5 g/L D -galactose.EGB761 was added to cultured cardiomyocytes at different concentration（5,10 and 20μg/mL,respectively） for 48 hours.Cell aging was determined by a senescentβ-galactosidase staining.Advanced glycation end products （AGEs） were quantitative analyzed by ELISA. The morphology and beat of cardiac myocytes were observed under an invert microscope.RESULTS： D-galactose resulted in a significantly increasedβ-galactosidase positive cardiomyocytes compared with the controls（75.6%±4.9%vs 17.15%±2.9%,P〈0.01）,concomited with elevated AGEs[（703＋32）vs（93±26） pg/mL, P〈0.01],abnormal cellular morphology and beat rate.EGB761 treated groups showed significant decrease in bothβ-galactosidase positive cells（57.7%±7.9%,49.7%±9.2%,34.0%±6.6%,P〈0.01） and AGEs levels[（404±33）, （357±25）,（249±77） pg/mL,respectively,all P value were less than 0.01],accompanied by relative normal morphology and beat rate.CONCLUSION： EGB761 has anti-aging actions on cardiomyocytes through inhibit non-enzymatic reactions.