杨正丽, 陶功华, 肖萍, 洪新宇. 5-氨基乙酰丙酸对持续性高糖致内皮细胞氧化损伤的干预作用研究[J]. 环境与职业医学, 2020, 37(7): 701-705, 718. DOI: 10.13213/j.cnki.jeom.2020.19863
引用本文: 杨正丽, 陶功华, 肖萍, 洪新宇. 5-氨基乙酰丙酸对持续性高糖致内皮细胞氧化损伤的干预作用研究[J]. 环境与职业医学, 2020, 37(7): 701-705, 718. DOI: 10.13213/j.cnki.jeom.2020.19863
YANG Zheng-li, TAO Gong-hua, XIAO Ping, HONG Xin-yu. Intervention of 5-aminolevulinic acid on oxidative damage of endothelial cells induced by persistent hyperglycemia[J]. Journal of Environmental and Occupational Medicine, 2020, 37(7): 701-705, 718. DOI: 10.13213/j.cnki.jeom.2020.19863
Citation: YANG Zheng-li, TAO Gong-hua, XIAO Ping, HONG Xin-yu. Intervention of 5-aminolevulinic acid on oxidative damage of endothelial cells induced by persistent hyperglycemia[J]. Journal of Environmental and Occupational Medicine, 2020, 37(7): 701-705, 718. DOI: 10.13213/j.cnki.jeom.2020.19863

5-氨基乙酰丙酸对持续性高糖致内皮细胞氧化损伤的干预作用研究

Intervention of 5-aminolevulinic acid on oxidative damage of endothelial cells induced by persistent hyperglycemia

  • 摘要: 背景

    5-氨基乙酰丙酸(5-ALA)是一种天然存在的氨基酸,广泛存在于植物和动物中。目前研究发现,5-ALA具有增强胰岛素敏感性和改善糖耐量水平的作用,而其在高糖诱导的内皮细胞损伤中的作用及其机制尚不清楚。

    目的

    探讨5-ALA在持续性高糖致人脐静脉内皮细胞(HUVECs)氧化损伤中的作用。

    方法

    设置阴性对照组、持续性高糖组、5-ALA干预组(1.0、10.0、50.0、100.0、200.0mmol·L-1)。用5-ALA预处理HUVECs 24 h,而后在细胞培养液中加入葡萄糖至终浓度为30 mmol·L-1,持续性高糖组细胞在含30 mmol·L-1葡萄糖的培养基中培养,阴性对照组细胞在正常培养基中培养,各培养48h。处理结束后,用CCK-8检测细胞活性,用相应试剂盒检测活性氧(ROS)水平、抗氧化酶总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量。

    结果

    与阴性对照组相比,持续性高糖组HUVECs活性降低(t=6.620,P < 0.01),HUVECs内ROS的水平增加(t=-10.671,P < 0.01),HUVECs内抗氧化酶T-SOD、CAT和GSH-Px的活性降低,MDA的含量增加(t=11.348、14.302、14.951、-12.562,均P < 0.01)。经5-ALA预处理后,与持续性高糖组相比,干预组HUVECs活性在不同程度上增加,其中50.0 mmol·L-1 5-ALA组增加最明显(t=-5.032,P < 0.01);HUVECs内ROS的水平降低,其中200 mmol·L-1 5-ALA组降低最明显(t=11.232,P < 0.01)。5-ALA可增加抗氧化酶的活性,并降低MDA的含量。对于T-SOD、GSH-Px和MDA而言,50.0 mmol·L-1 5-ALA干预效果最佳(t=-8.562、-14.273、12.526,均P < 0.01);对于CAT而言,100.0mmol·L-1 5-ALA干预组升高最明显(t=-21.467,P < 0.01)。

    结论

    持续性高糖可增加内皮细胞内ROS的水平,降低T-SOD、CAT和GSH-Px的活性和增加MDA的含量,而5-ALA在一定程度上减轻持续性高糖引起的氧化损伤。

     

    Abstract: Background

    5-aminolevulinic acid (5-ALA) is a naturally occurring amino acid found in plants and animals. Current studies have found that 5-ALA can enhance insulin sensitivity and improve glucose tolerance, but its role and mechanism in hyperglycemia-induced endothelial cell injury are still unclear.

    Objective

    This experiment investigates the role of 5-ALA in the oxidative injury of human umbilical vein endothelial cells (HUVECs) induced by persistent hyperglycemia.

    Methods

    The experiment was designed to include a negative control group, a persistent hyperglycemia group, and 1.0, 10.0, 50.0, 100.0, and 200.0 mmol·L-1 5-ALA intervention groups. The treatment protocol was that the cells in the 5-ALA intervention groups were pretreated with 5-ALA for 24 h, and then glucose was added to the cell culture solution to a final concentration of 30 mmol·L-1 for 48 h; the cells in the persistent hyperglycemia group were cultured in a medium with a glucose concentration of 30 mmol·L-1 for 48 h; the cells in the negative control group were cultured in regular medium for 48h. After the treatment, the cell viability was detected by CCK-8, and the reactive oxygen species (ROS) level, the activities of intracellular antioxidant enzymestotal superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) content were detected with corresponding kits.

    Results

    Compared with the negative control group, the persistent hyperglycemia group showed decreased HUVECs viability (t=6.620, P < 0.01), increased ROS level (t=-10.671, P < 0.01), decreased activities of antioxidant enzymes T-SOD, CAT, and GSH-Px, and increased content of MDA (t=11.348, 14.302, 14.951, and -12.562, respectively, P < 0.01). After the designed 5-ALA intervention, compared with the persistent hyperglycemia group, the intervention groups showed increased HUVECs viability to varying degrees, of which the 50.0 mmol·L-1 5-ALA intervention group was the most obvious (t=-5.032, P < 0.01); the intervention groups showed reduced ROS levels in HUVECs to different degrees, of which the 200 mmol·L-1 5-ALA group had the sharpest reduction (t=11.232, P < 0.01). The 5-ALA intervention also increased the antioxidant enzyme activities and reduced the MDA content. For T-SOD, GSH-Px, and MDA, the effect in the 50.0 mmol·L-1 5-ALA intervention was the best (t=-8.562, -14.273, and 12.526, respectively, P < 0.01); for CAT, the 100.0 mmol·L-1 5-ALA intervention had the best effect (t=-21.467, P < 0.01).

    Conclusion

    Persistent hyperglycemia can increase the level of ROS in endothelial cells, reduce the activities of T-SOD, CAT and GSH-Px, and increase the content of MDA; 5-ALA can extenuate the oxidative injury induced by persistent hyperglycemia to a certain extent.

     

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