D-半乳糖诱导心肌细胞非酶糖基化对钙调节蛋白的影响  

Influence of non-enzymatic glycosylation induced by D-galactose on the expression of calmodulin in cultured cardiomyocytes

作　　者：郭长青[1] 郭妍[1] 陈相健[2] 

机构地区：[1]南京医科大学第一附属医院老年医学科,江苏省南京市210029 [2]南京医科大学第一附属医院心肌研究所,江苏省南京市210029

出　　处：《实用老年医学》2010年第3期200-202,212,共4页Practical Geriatrics

基　　金：国家自然科学基金项目(30772781)

摘　　要：目的研究D-半乳糖诱导衰老的心肌细胞中的非酶糖基化反应,及其对心肌细胞内钙调节蛋白水平的影响。方法实验用5g/LD-半乳糖干预培养心肌细胞建立衰老模型。显微镜下观察心肌细胞的形态和搏动情况。以β-半乳糖苷酶染色观察细胞衰老,ELISA法检测心肌细胞晚期糖基化终产物(AGEs)的含量,采用western blot检测肌浆网Ca2+-ATP酶(SERCA,心脏主要是SERCA2a)及受磷蛋白(PLN)的含量。结果 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高于正常对照组[(75.6±4.9)%和(17.15±2.9)%,P<0.01)];细胞内AGEs较正常对照组明显增高[(702.58±32.16)pg/ml和(93.22±26.14)pg/ml,P<0.01];SER-CA2a含量明显下降与正常对照组比值为0.39±0.05(P<0.01);PLN含量明显下降与正常对照组比值为0.27±0.03(P<0.01);同时伴有心肌细胞形态和搏动的异常。结论 D-半乳糖可通过心肌细胞的非酶糖基化诱导的心肌细胞老化,同时降低心肌细胞内的SERCA2a、PLN蛋白含量,影响心肌细胞的舒缩活动。Objective To investigate the non-enzymatic reactions effects of D-galactose induced on cardiomyocytes aging and the influence on the calmodulin. Methods The cultured neonatal rat cardiomyocytes were induced by D-galactose（5g/L）.Cell aging was detected by β-galactosidase staining and the quantity analysis of advanced glycation end pro-ducts（AGEs） by ELISA. The appearance and beating of cardiac myocytes were observed under an invert microscope. Levels of essential sarcoplasmic reticulum Ca^2＋-ATPase （SERCA2a） and phospholamban （PLN）were assessed by Western blotting. Results D-galactose decreased trend to induce significantly increased positive β-galactosidase staining （75.6±4.9）% in cardiac myocytes,concomitant with enhanced AGEs （702.58±32.16 pg/ml,P〈0.01）. SERCA2a in contrast with the normal ratio is 0.39±0.05（P〈0.01）,and PLN in contrast with normal ratio is 0.27±0.03（P〈0.01）. Abnormal cardiomyocyte modality and beat rate can also be observed. Conclusions D-galactose may induce cardiomyocyte cell aging through non-enzymatic reactions,decrease the expression of SERCA2a and PLN,and thus influence the appearance and beat of cardiac myocytes.

关 键 词：晚期糖基化终产物 心肌老化 肌浆网Ca2+-ATP酶 受磷蛋白 

分 类 号：Q255[生物学—细胞生物学]