Saikosaponin-D attenuates heat stress-induced oxidative damage in LLC-PK1 cells by increasing the expression of anti-oxidant enzymes and HSP72.
Saikosaponin-D attenuates heat stress-induced oxidative damage in LLC-PK1 cells by increasing the expression of anti-oxidant enzymes and HSP72.
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DOI：
10.1142/s0192415x14500797
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发表时间：
2014-09
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影响因子：
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通讯作者：
Baoren Zhang;Xiaodan Guo;Jian-Wei Chen;Yuan Zhao;X. Cong;Zhongling Jiang;Rongfeng Cao;Kai Cui;Shan-song Gao;W. Tian
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文献类型：
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作者：
Baoren Zhang;Xiaodan Guo;Jian-Wei Chen;Yuan Zhao;X. Cong;Zhongling Jiang;Rongfeng Cao;Kai Cui;Shan-song Gao;W. Tian
热应激刺激活性氧（ROS）的产生，导致肾脏的氧化损伤。本研究旨在阐明柴胡皂苷d（Saikosaponin-d，SSd）对热应激猪肾近端小管（LLC-PK 1）细胞氧化损伤的保护机制。如3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑（MTT）测定所证明的，SSd单独在1或3 μg/mL浓度下没有细胞毒性。为了评估SSd对热应激诱导的细胞损伤的影响，LLC-PK 1细胞用各种浓度的SSd预处理，在42°C下热应激1小时，然后返回到37°C持续9小时。DNA Ladder和MTT实验表明，与未处理的细胞相比，SSd有助于防止热应激诱导的细胞损伤。此外，与对照组相比，SSd预处理增加超氧化物歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GPx）的活性，但降低丙二醛（MDA）的浓度呈剂量依赖性。实时荧光定量PCR和Western blot分析结果表明，SSd可显著提高铜锌超氧化物歧化酶（SOD-1）、过氧化氢酶（CAT）、GPx-1和热休克蛋白72（HSP 72）在mRNA和蛋白水平的表达。总之，这些结果是第一次证明，SSd通过调节LLC-PK 1细胞中抗氧化酶和HSP 72的活性来改善热应激诱导的氧化损伤。
Heat stress stimulates the production of reactive oxygen species (ROS), which cause oxidative damage in the kidney. This study clarifies the mechanism by which saikosaponin-d (SSd), which is extracted from the roots of Bupleurum falcatum L, protects heat-stressed pig kidney proximal tubular (LLC-PK1) cells against oxidative damage. SSd alone is not cytotoxic at concentrations of 1 or 3 μg/mL as demonstrated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To assess the effects of SSd on heat stress-induced cellular damage, LLC-PK1 cells were pretreated with various concentrations of SSd, heat stressed at 42°C for 1 h, and then returned to 37°C for 9 h. DNA ladder and MTT assays demonstrated that SSd helped to prevent heat stress-induced cellular damage when compared to untreated cells. Additionally, pretreatment with SSd increased the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) but decreased the concentration of malondialdehyde (MDA) in a dose-dependent manner when compared to controls. Furthermore, real-time PCR and Western blot analysis demonstrated that SSd significantly increased the expression of copper and zinc superoxide dismutase (SOD-1), CAT, GPx-1 and heat shock protein 72 (HSP72) at both the mRNA and protein levels. In conclusion, these results are the first to demonstrate that SSd ameliorates heat stress-induced oxidative damage by modulating the activity of anti-oxidant enzymes and HSP72 in LLC-PK1 cells.