Oligomeric proanthocyanidins protect against HK-2 cell injury induced by oxalate and calcium oxalate monohydrate crystals
Oligomeric proanthocyanidins protect against HK-2 cell injury induced by oxalate and calcium oxalate monohydrate crystals
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DOI：
10.1007/s00240-015-0826-9
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发表时间：
2016-06-01
期刊：
影响因子：
3.1
通讯作者：
Yang, Yong
中科院分区：
文献类型：
--
作者：
Wang, Shuo;Du, Peng;Yang, Yong
本研究旨在检测抗氧化剂低聚原花青素(OPC)对一水草酸钙晶体(COM)对HK-2细胞毒性的保护作用。实验分为4组：阴性对照组、阳性对照组(COM+草酸盐)、OPCS组(OPCS+COM+草酸盐)、VitE组(VitE+COM+草酸盐)。分别培养HK-2细胞4、8、12、24 h，用四甲基偶氮唑盐比色法检测HK-2细胞活性。用Na+/K+-ATP酶活性评价细胞损伤程度。以培养上清液中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性为指标评价过氧化程度。在透射电子显微镜下观察各组HK-2细胞暴露4h和12h后的形态变化。检测OPC和VitE对草酸和COM染毒细胞的影响。经草酸盐和COM晶体处理后，细胞活性、Na+/K+-ATP酶和SOD酶活性显著降低，培养液中丙二醛含量显著升高。OPCS组：与阳性对照组相比，OPCS组细胞活性、超氧化物歧化酶和Na+/K+-ATP酶活性显著升高，丙二醛含量显著降低。VitE组：与阳性对照组相比，加入VitE后，HK-2细胞的Na+/K+-ATP酶活性和Na+/K+-ATP酶活性无明显变化，而SOD活性恢复，丙二醛含量显著降低，经高浓度COM晶体和草酸盐处理后，细胞形态结构发生明显变化。加入OPC或VitE后，细胞质内空泡形成、核膜溶解、核仁消失的细胞数较阳性对照组减少。OPCs组的形态结构改变较VitE组轻。OPC和维生素E的应用可以防止草酸和COM介导的过氧化损伤，恢复细胞内抗氧化酶活性。OPC提供的保护作用强于维生素E。
The purpose of the study was to test whether the antioxidants oligomeric proanthocyanidins (OPCs) could provide protection against oxalate and calcium oxalate monohydrate crystals (COM) toxicity in HK-2 cells. Four groups were chosen for the study: negative control group, positive control group (COM + oxalate), OPCs group (OPCs + COM + oxalate), Vit E group (Vit E + COM + oxalate). HK-2 cells were exposed for 4, 8, 12 and 24 h. The activity of HK-2 cell was assessed by MTT. Cellular injury was assessed by activity of Na+/K+ ATP enzyme. Peroxidation level was assessed by malondialdehyde (MDA) content in medium and activity of superoxide dismutase (SOD). Morphological changes of HK-2 cell after exposed for 4 and 12 h in each group were observed under Transmission electron microscope (TEM). The effects of OPCs and VitE on oxalate- and COM-exposed cells were tested. After exposed to oxalate and COM crystals, activity of cells, Na+/K+ ATP enzyme and SOD enzyme showed a significant reduction, and MDA content in medium was significantly increased. OPCs group: the addition of OPCs significantly increased activity of cell, SOD and Na+/K+ ATP enzyme while MDA content was significantly decreased compared with the positive control group. VitE group: compared with the positive control group, activity of HK-2 cell, Na+/K+ ATP enzyme was not significantly changed while SOD activity was restored, and MDA content was significantly decreased after the addition of Vit E. Morphological structure of HK-2 cell was extremely changed as observed under TEM after exposure to high level of COM crystals and oxalate. After the addition of OPCs or Vit E, amounts of cells with vacuoles formed in cytoplasms, karyotheca dissolved and nucleolus disappeared were less than in positive control group. The morphological structure changing in OPCs group was slighter than that in Vit E group. OPCs and vitamin E administration may prevent oxalate- and COM-mediated peroxidative injury, restoring intracellular antioxidant enzyme activity. The protection rendered by OPCs was greater than that of vitamin E.