微管干预剂对缺氧心肌细胞糖酵解途径关键酶的影响

目的 了解微管干预剂对缺氧心肌细胞糖酵解途径关键酶的影响. 方法体外培养心肌细胞,分为单纯缺氧组、缺氧+微管解聚剂(秋水仙碱)组、缺氧+低浓度微管稳定剂组、缺氧+中浓度微管稳定剂组、缺氧+高浓度微管稳定剂组,后3组加入的微管稳定剂为紫杉醇,浓度分别为5、10、15 μmol/L.采用激光共聚焦显微镜观察微管形态学变化,检测细胞活力及己糖激酶、丙酮酸激酶、磷酸果糖激酶、乳酸脱氢酶的活性. 结果缺氧+微管解聚剂组和缺氧+高浓度微管稳定剂组心肌细胞微管结构破坏显著,细胞活力亦明显下降缺氧1.0 h,2组细胞活力分别为(89.99±3.47)%、(84.56±6.61)%,明显低于单纯缺氧组(97.44±1.76)%(P<0.01)],前3种酶的活性亦明显降低;缺氧+低浓度微管稳定剂组前3种酶的活性与单纯缺氧组基本近似;缺氧+中浓度微管稳定剂组微管结构损伤显著轻于其他组,前3种酶的活性于缺氧6.0 h内高于单纯缺氧组.缺氧后各组乳酸脱氢酶活性升高. 结论适当浓度的微管稳定剂能明显减轻微管结构损伤,促使缺氧早期心肌细胞糖酵解酶活性增强.

The influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia

OBJECTIVE: To investigate the influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia. METHODS: The primary passage of cultured myocardial cells from neonatal rats were divided into A group (with hypoxia), B group (with hypoxia and administration of l0 micromol/L colchicine), C group (with hypoxia and administration of 5 micromol/L taxol), D group (with hypoxia and administration of 10 micromol/L taxol), E group (with hypoxia and administration of 15 micromol/L taxol). The morphology of microtubule was observed with laser scanning microscope (LSM). The cell vitality was assayed by cell counting kit (CCK). The activities of hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) were assayed with colorimetry. RESULTS: In group B and E, the microtubule structure was damaged heavily, and the cell vitality was decreased significantly The cell vitality was (89.99 +/- 3.47)% in B group and (84.56 +/- 6.61)% in E group, respectively, at 1.0 post hypoxia hour (PHH), and hoth values were obviously lower than that in A group (97.44 +/- 1.76)%, P < 0.01]. The HK, PK and PFK activities decreased obviously. The activities of HK, PK and PFK in group C were similar to those of the A group. Compared with that in other groups, the degree of damage of microtubule structure in D group was milden. The activities of HK, PK and PFK in D group during 0.5 - 6.0 PHH were significantly higher than those in A group. The activity of LDH in each group was increased after hypoxia. CONCLUSION: Proper concentration of microtubule-stabilizing drugs can alleviate the damages to microtubule structure, and enhance the activity of glycolytic key enzymes of myocardial cells at early stage of hypoxia.