半胱胺对肝细胞代谢油酸过程中ROS生成和ATPase活性的影响

EFFECT OF CYSTEAMINE ON THE LEVEL OF ROS AND ATPASE IN THE METABOLISM OF OLEIC ACID IN HEPATOCYTES

目的研究了半胱胺(CS)对油酸(OA)代谢过程中自由基(ROS)生成和ATP酶(ATPase)活性的影响。方法通过培养体外小鼠原代肝细胞,添加不同浓度的OA作用24h,通过luminol化学发光法,TBA法、DTNB法、无机磷比色法分别测定ROS、丙二醛(MDA)、谷胱甘肽(GSH)和Na+-K+-ATP酶活性;选取OA(0,150,250μM/L)三个梯度,添加CS(0,50,100,150,200,250,300,350,400,450,500μM/L),测定相同的指标。结果添加OA大于150μM/L时,ROS、MDA生成量急剧升高,ATPase活性显著降低(P<0.05)。添加CS能提高GSH,减少ROS和MDA的生成,提高ATPase的活性,且作用的效果显著(P<0.05)。但大量添加CS(350-500μM/L)则效果相反。结论大量添加OA会引起肝细胞氧化应激;添加CS能清除ROS,提高ATPase活性;并伴随OA添加量的增加,CS的最佳需要量也增加。

Objective To study the effect of cysteamine （CS） on the level of ROS and ATP during the metabolism of oleic acid （OA）. Methods Primary hepatic cells were treated with OA （0, 50, 150, 200, 250μM/L） for 48h; levels of MDA, GSH and ROS were measured in the hepatic cells by chemilumi- nescence assay using TBA, DTNB and luminal respectively; the activity of the Na＋-K＋-ATPase was tested by inorganic phosphorus spectrophotometer. In addition, the hepatic cells treated with OA （0, 150, 250μM/L） were analysed in the presense of CS （0, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500μM/L） for 24h. Results High concentration of OA （150--250μM/L） induced hepatocyte oxidation accompanied by marked increase in ROS and MDA levels and decrease in the activity of ATPase （P〈0. 05）. CS could raise GSH and activity of ATPase, and reduce the production of MDA and ROS. However, high concentrations of CS （350-500μM/L） acted inversely. Conclusion High concentrations of OA can induce lipid oxidation; CS can inhibit it by eliminating ROS and elevating ATPase activity in a dose-dependent manner.