锌补充通过上调金属硫蛋白缓解OTA 诱导的 MDCK 细胞氧化应激和凋亡。
Zinc supplementation alleviates OTA-induced oxidative stress and apoptosis in MDCK cells by up-regulating metallothioneins.
机构信息
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China; Institute of Nutritional and Metabolic Disorders in Domestic Animals and Fowls, Nanjing Agricultural University, Nanjing 210095, China; MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine Nanjing Agricultural University Nanjing, China.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China; Institute of Nutritional and Metabolic Disorders in Domestic Animals and Fowls, Nanjing Agricultural University, Nanjing 210095, China; MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine Nanjing Agricultural University Nanjing, China.
出版信息
Life Sci. 2019 Oct 1;234:116735. doi: 10.1016/j.lfs.2019.116735. Epub 2019 Aug 5.
AIMS
The present study was to investigate the protective effects of Zn supplementation in OTA-induced apoptosis of Madin-Darby canine kidney (MDCK) epithelial cells and explore the potential mechanisms. Aiming to provides a new insight into the treatment strategy of OTA-induced nephrotoxicity by nutritional regulation.
MAIN METHODS
Initially, through MTT and LDH assay revealed that Zn supplementation significantly suppressed OTA-induced cytotoxicity in MDCK cells. Then, the production of reactive oxygen species (ROS) was detected by using a DCFH-DA assay. Annexin V-FITC/PI, Hoechst 33258 staining and Flow cytometry were used to detect the apoptosis. The expressions of apoptosis-related molecules were determined by RT-PCR, Western blotting. Interestingly, OTA treatment slightly increased the levels of Metallothionein-1 (MT-1) and Metallothionein-2 (MT-2) by using RT-PCR, Western blotting assay; while Zn supplementation further improved the increase of MT-1 and MT-2 induced by OTA. However, the inhibitive effects of Zn supplementation were significantly blocked after double knockdown of MT-1 and MT-2 by using Small Interfering RNA (siRNA) Transfection method.
KEY FINDINGS
Our study provides supportive data for the potential roles of Zn in reducing OTA-induced oxidative stress and apoptosis in MDCK cells.
SIGNIFICANCE
Zn is one of the key structural components of many proteins, which plays an important role in several physiological processes such as cell survival and apoptosis. This metal is expected to contribute to the conservative and adjuvant treatment of kidney disease and should therefore be investigated further.
目的
本研究旨在探讨锌补充对OTA 诱导的 Madin-Darby 犬肾（MDCK）上皮细胞凋亡的保护作用，并探讨其潜在机制。旨在为通过营养调节治疗 OTA 诱导的肾毒性提供新的思路。
主要方法
首先，通过 MTT 和 LDH 测定法发现，锌补充可显著抑制 OTA 诱导的 MDCK 细胞毒性。然后，通过 DCFH-DA 测定法检测活性氧（ROS）的产生。使用 Annexin V-FITC/PI、Hoechst 33258 染色和流式细胞术检测细胞凋亡。通过 RT-PCR、Western blot 测定法测定凋亡相关分子的表达。有趣的是，通过 RT-PCR、Western blot 测定法发现，OTA 处理轻度增加了 Metallothionein-1（MT-1）和 Metallothionein-2（MT-2）的水平；而锌补充进一步提高了 OTA 诱导的 MT-1 和 MT-2 的增加。然而，在用小干扰 RNA（siRNA）转染法进行 MT-1 和 MT-2 的双重敲低后，锌补充的抑制作用显著被阻断。
主要发现
我们的研究为锌在降低 MDCK 细胞中 OTA 诱导的氧化应激和细胞凋亡中的潜在作用提供了支持性数据。
意义
锌是许多蛋白质的关键结构成分之一，在细胞存活和凋亡等多种生理过程中发挥重要作用。这种金属有望为肾脏疾病的保守和辅助治疗做出贡献，因此应进一步研究。
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