低氧对新生猪G_1/S肾小管上皮细胞能量代谢影响的特点

Effect of Hypoxia on Energy Metabolism of G_1/S Renal Epithelial Cells of Piglets

目的 探讨窒息早期所致低氧对围生期 G1 / S界面肾小管上皮细胞 (RTEs)能量代谢的影响。方法 应用二次过量胸苷同步化法获取 G1 / S界面细胞 (应用流式细胞术进行鉴定 )。将 RTEs分为三组 :采用氰化钠(Na CN)进行 G1 / S界面 RTEs缺氧细胞模型 (缺氧组 ) ,同时设立预防组 (氰化钠 +斑蟊素 )和未处理对照组。反相高效液相检测缺氧前后及恢复 6 0 m in、12 0 min、180 min各组细胞内 ATP含量的变化。结果 二次胸苷同步化法可获取约 (99.5 2± 0 .83) %的 G1 / S界面细胞 ;缺氧组在缺氧后 0 m in和恢复 6 0 m in及 12 0 m in时 RTEs ATP含量均高于预防组和对照组 (P<0 .0 5 ) ,而在恢复 180 min时细胞内 ATP的含量均低于预防组和对照组 (P<0 .0 5 )。结论 体外化学缺氧对 G1 / S界面

Objective To determine whether effect of energy metabolism on G 1/S renal tubular epithelial cells(RTEs) is strongly associated with anoxemia. Methods Neonatal pig's RTEs were synchronized by the Excess Thymidine Block technique and the distribution of G 1/S phases was determined by flow cytometry (FACS). The hypoxic-cell model of G 1-phase RTEs was established by exposure of the RTEs to sodium cyanide (NaCN), and the preventive (NaCN+Cantharidin) group and un-treated control group were set up at the same time. The ATP levels in three RTEs groups were measured by high-performance liquid chromatography. Results FACS measurements revealed that (99.52±0.83)% of the neonatal RTEs were at G 1/S transition. The concentration of ATP in the RTEs of hypoxic group at 0, 60, 120 minutes after with NaCN treatment for one hour was higher than that of the preventive group and un-treated control group (P<0.05). At 180 minutes after NaCN treatment, the RTEs' ATP in hypoxic group was significantly reduced as compared with those of the preventive and un-treated control groups (P<0.05). Conclusion Abnormal energy metabolism in newborn piglet's RTEs is significantly associated with anoxia that may further interfere with the advance of RTEs' cell cycle and Cantharidin can regulate the ATP metabolism of hypoxic RTEs.