研究了氧化应激对肾细胞-Vero细胞的损伤及EGCG的保护作用。用MTT、吖啶橙染色和DNA凝胶电泳等方法研究了H2O2和Cr6+应激对Vero细胞的损伤,及EGCG对凋亡细胞的保护作用及其机理。结果表明,H2O2和Cr6+剂量效应地抑制Vero细胞活性,IC50分别为175.6和9.8mg·L-1;其中50 mg·L-1 H2O2和400 mg·L-1/2h Cr6+可诱导Vero细胞凋亡。0~60 mg·L-1 EGCG有效抑制H2O2和Cr6+应激引起的细胞活性下降,且40 mg·L-1 EGCG显著抑制细胞凋亡。对于Cr6+所诱导的细胞凋亡,EGCG的保护作用与EDTA和Vc的协同作用效果相当,表明清除活性氧和络合金属离子都有助于减轻Cr6+应激损伤。可见,EGCG通过清除活性氧和络合离子有效地保护了肾细胞免受应激损伤,这对易遭受活性氧损伤的肾脏无疑具有一定的临床价值。
This study was conducted to identify the protection of antioxidant--epigallocatechin-3-gallate(EGCG) to the renal damage by oxidant-stress. H2O2 and Cr6+ were used to induce apoptosis of renal cells-----Vero cells in vitro. With these models of cell apoptosis, the effect of EGCG on apoptosis was investigated through Staining and DNA electrophoresis. As the results, H2O2 and Cr6+ decreased the viability of Vero cells in a dose-dependent manner, and the IC50 was 175.6 and 9.8 mg·L- respectively. Both 50 mg·L-1 H2O2 and 400 mg·L-/2h Cr6+ caused the apoptosis of Vero cells. 40 mg·L- EGCG markedly inhibited the apoptosis of Vero cells, and the effect was the same as that of Vc and EDTA in combination, indicating the simultaneous effect of antioxidation and chelating. So, EGCG protected Vero cells from damage through both ROS scavenging and ion-chelating, and this kind of effect is valuable to kidney usually exposed to ROS in clinic.
[1] 王海燕. 肾脏病学[M]. 人民卫生出版社,北京,1996(第二版).
[2] 王叔兰. 活性氧自由基损伤与肾小球疾病[J]. 中级医刊,1996,31(6):19-21.
[3] Sonal A, Kajal M, Smita K, Rao CV.A subtoxic interactive toxicity study of ethanol and chromium in male Wistar rats[J]. Alcohol, 2001, 23: 99-108.
[4] Dartsch PC, Hildenbrand S, Kimmel R, Schmahl FW.Investigations on the nephrotoxicity and hepatotoxicity of trivalent and hexavalent chromium compounds[J]. Occupational Health and Industrial Medicine, 1999, 40(1):12.
[5] Jiang B, Zhang YL, Zhou DY.Common Experimental Method of molecule Biology (分子生物学常用实验方法)[M]. Beijing: People’s Surgeon Publishing House,1996. 45.
[6] Zhao WH.Apoptosis of Cells(细胞凋亡)[M]. Henan: Medical University’s Publishing House in Henan, 1997. 132-133.
[7] Suzuki Y J, Forman HJ,Sevanian A.Oxidants as stimulators of signal transduction. Free Radic. Biol. Med, 1997, 22:269-285.
[8] Leonard S, Wang SW, Zang LY, Vallyathan VC and Shi XL. Role of molecular oxygen in the generation of hydroxyl and superoxide anion radicals during enzymatic Cr(VI) reduciton and its implication to Cr(VI)-induced carcinogenesis[J]. Journal of Enviromental Pathology, Toxicology and Oncology, 2000, 19(1&2):49-60.
浙公网安备 33019902000101号 
