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为观察葡萄糖和胰岛素对血管平滑肌细胞磷脂酶活性的影响 ,在培养基中分别加入一定浓度的葡萄糖或 (和 )胰岛素与大鼠血管平滑肌细胞共同孵育 72h后 ,分别测定培养上清液中磷脂酶A2 活性和Na+-K+依赖式ATP酶活性 ,磷脂酶A2 活性检测采用 [3 H]—油酸标记法 ,Na+-K+依赖式ATP酶活性检测采用以ATP·2Na为底物的比色法。结果发现大鼠血管平滑肌细胞暴露于高糖或 (和 )高胰岛素环境中 ,上清液中磷脂酶A2 活性增加 ,而Na+-K+依赖式ATP酶活性下降 ,并在一定浓度范围内呈剂量依赖性。葡萄糖浓度由 5 .6mmol L增加到 33.0mmol L ,上清液中磷脂酶A2 活性由 ( 4.2± 0 .2 )× 10 1 2 (min·L)增加到 ( 9.2± 0 .2 )× 10 1 2 (min·L) ;而Na+-K+依赖式ATP酶活性由 4.8± 1.0 2 μmol (g·min)下降到 2 .0 6± 0 .99μmol (g·min)。胰岛素浓度由 5 .0Mu L增加到 10 0 0Mu L ,上清液中磷脂酶A2 活性由 ( 3 .8± 0 .4)× 10 1 2 (min·L)增加到 ( 11.8± 0 .6 )× 10 1 2 (min·L) ;而Na+-K+依赖式ATP酶活性由 4.0± 1.6 8μmol (g·min)下降到 2 .19± 1.0 2 μmol (g·min) ,且进一步发现这一效应在作用 2 4h后才显著 ,高糖和高胰岛素有协同作用。结果提示磷脂酶A2 活性增高与糖尿病时动脉粥样硬化形成有关 ,?
In order to observe the effect of glucose and insulin on the phospholipase activity of vascular smooth muscle cells, a certain concentration of glucose or (and) insulin and rat vascular smooth muscle cells were incubated for 72 hours in culture medium respectively, and the phospholipase A2 activity and Na + -K + -dependent ATPase activity, phospholipase A2 activity by [3 H] -oleic acid labeling method and Na + -K + -dependent ATPase activity assay by ATP · 2Na substrate assay. The results showed that rat vascular smooth muscle cells exposed to high glucose or (and) high insulin environment, the phospholipase A2 activity in the supernatant increased, while the Na + -K + -dependent ATPase activity decreased and in a certain concentration range was dose-dependent Sex. The glucose concentration increased from 5.6 mmol L to 33.0 mmol L, and the phospholipase A2 activity in the supernatant increased from (4.2 ± 0.2) × 10 12 (min · L) to (9.2 ± 0.2) × 10 1 2 (min · L), while Na + -K + -dependent ATPase activity decreased from 4.8 ± 1.0 2 μmol (g · min) to 2.06 ± 0.99 μmol (g · min). Insulin concentration increased from 5.0 μM to 1.0 μM L, phospholipase A2 activity in supernatant increased from (3.8 ± 0.4) × 10 12 (min · L) to (11.8 ± 0.6) × 10 1 2 (min · L), while the activity of Na + -K + -dependent ATPase decreased from 4.0 ± 1.6 8μmol (g · min) to 2.19 ± 1.0 2 μmol (g · min), and this effect was further found In the role of 24 h after significant, high glucose and insulin have synergistic effects. The results suggest that increased phospholipase A2 activity is associated with the development of atherosclerosis in diabetes.