Wang Shuo, Huang Xiao-bo, Xu Qing-quan, Ye Hai-yun, Ma Kai, Wang Xiao-feng
Department of Urology, Peking University People's Hospital, Beijing 100034, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2013 Aug 18;45(4):567-74.
To test whether the antioxidants tea polyphenol (TP) can provide protection against oxalate and calcium oxalate monohydrate (COM) crystals toxicity in HK-2 cells.
Four groups were chosen for the study: Negative control group, positive control group (COM+oxalate), TP group (TP+COM+oxalate), VitE group (vitamin E+COM+oxalate). HK-2 cells were exposed for 4, 8, 12 and 24 h. The viability of the cells was assessed by MTT. The cellular injury was assessed by the concentration of lactate dehydrogenase (LDH), hydrogen peroxide and viability of Na+/K+ ATP enzyme. The peroxidation level was assessed by malondialdehyde (MDA) content and viability of superoxide dismutase (SOD). The morphological changes of HK-2 cells after being exposed for 4 and 12 h in each group were observed by transmission electron microscope (TEM).
The effects of TP and vitamin E on oxalate and COM exposed cells were tested. The HK-2 cells exposed to oxalate and COM showed a significant reduction in viability of cells, Na+/K+ ATPase and SOD. LDH release, MDA content and concentration of H2O2 were significantly increased. In TP group, the addition of TP significantly increased viability of cells, activity of Na+/K+ ATPase and SOD while LDH release, MDA content and concentration of H2O2 were significant decreased compared with the positive control group. In the Vitamn E group, compared with the positive control group, viability of cells, and activity of Na+/K+ ATPase were not significantly changed and after addition of vitamin E, SOD activity was restored, LDH release, MDA content and concentration of H2O2 were significant decreased compared with the positive control group. The morphological changes of HK-2 cells were observed by TEM in the positive control group, TP group and VitE group. In the VitE and TP groups, the amount of the cells with vacuoles formed in kytoplasms, mitochondria swelling, karyotheca dissolved and nucleolus disappearing were less than in the positive group. The morphological changes in the TP group were less than in the VitE group.
TP and vitamin E administration may prevent oxalate and COM mediated peroxidative injury and restore intracellular antioxidant enzyme activity. The protection rendered by TP was greater than that of vitamin E.
检测抗氧化剂茶多酚(TP)是否能保护HK-2细胞免受草酸盐和一水合草酸钙(COM)晶体毒性的影响。
研究选取四组:阴性对照组、阳性对照组(COM + 草酸盐)、TP组(TP + COM + 草酸盐)、维生素E组(维生素E + COM + 草酸盐)。将HK-2细胞暴露4、8、12和24小时。通过MTT法评估细胞活力。通过乳酸脱氢酶(LDH)浓度、过氧化氢和Na+/K+ ATP酶活力评估细胞损伤。通过丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力评估过氧化水平。用透射电子显微镜(TEM)观察每组HK-2细胞在暴露4小时和12小时后的形态变化。
检测了TP和维生素E对暴露于草酸盐和COM的细胞的影响。暴露于草酸盐和COM的HK-2细胞显示细胞活力、Na+/K+ ATP酶和SOD显著降低。LDH释放、MDA含量和H2O2浓度显著增加。在TP组中,与阳性对照组相比,添加TP显著提高了细胞活力、Na+/K+ ATP酶活性和SOD活性,而LDH释放、MDA含量和H2O2浓度显著降低。在维生素E组中,与阳性对照组相比,细胞活力和Na+/K+ ATP酶活性没有显著变化,添加维生素E后,SOD活性恢复,与阳性对照组相比,LDH释放、MDA含量和H2O2浓度显著降低。在阳性对照组、TP组和维生素E组中用TEM观察HK-2细胞的形态变化。在维生素E组和TP组中,细胞质中形成空泡、线粒体肿胀、核膜溶解和核仁消失的细胞数量少于阳性组。TP组的形态变化小于维生素E组。
给予TP和维生素E可预防草酸盐和COM介导的过氧化损伤并恢复细胞内抗氧化酶活性。TP提供的保护作用大于维生素E。