Park C S, Jang H J, Lee J H, Oh M Y, Kim H J
Department of Surgery, Ulsan University, College of Medicine, Gangneung Asan Hospital, Gangneung, South Korea.
Department of Surgery, Ulsan University, College of Medicine, Gangneung Asan Hospital, Gangneung, South Korea.
Transplant Proc. 2018 Nov;50(9):2854-2859. doi: 10.1016/j.transproceed.2018.03.031. Epub 2018 Mar 16.
Calcineurin inhibitors are effective immunosuppressive agents, but associated adverse effects such as nephrotoxicity may limit efficacy. Tacrolimus (FK506) is an immunosuppressive drug used mainly to lower the risk of organ rejection after allogeneic organ transplant. Adverse effects of FK-506 can prompt patients to end treatment despite the efficacy. In the present study, we investigated the protective effect and mechanism of tetrahydrocurcumin (THC) on FK506-induced renal damage, apoptosis, and oxidative stress to evaluate its possible use for kidney protection.
The effect of THC on FK506-induced kidney cell damage was investigated in LLC-PK1 cells. LLC-PK1 cells were pretreated with THC at concentrations of dose for 2 hours followed by addition of FK506 for 24 hours. LLC-PK1 cells were treated with FK506 and THC, and cell viability and glutathione was measured. The number of apoptotic cells was measured using an annexin V/propidium iodide staining with flow cytometry. The effect of apoptosis by THC in LLC-PK1 cells was determined by measuring the caspase-9, caspase-3, B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein levels using Western blotting analyses.
FK506-induced LLC-PK1 renal cell damage was markedly ameliorated by THC treatment. THC protected LLC-PK1 cells by preventing FK506-induced glutathione decrease. THC protects against FK506-induced apoptosis in LLC-PK1 cells. Apoptosis was significantly decreased, and Bcl-2 was elevated in the THC-treated group. Bcl-2-associated X protein, caspase-3, and caspase-9 were decreased in the THC-treated group.
These results collectively provide therapeutic evidence that THC ameliorates the FK506-induced renal damage via antioxidant effect and apoptosis inhibition.
钙调神经磷酸酶抑制剂是有效的免疫抑制剂，但诸如肾毒性等相关不良反应可能会限制其疗效。他克莫司（FK506）是一种免疫抑制药物，主要用于降低同种异体器官移植后器官排斥的风险。尽管FK-506有疗效，但其不良反应可能促使患者终止治疗。在本研究中，我们研究了四氢姜黄素（THC）对FK506诱导的肾损伤、细胞凋亡和氧化应激的保护作用及机制，以评估其在肾脏保护方面的潜在用途。
在LLC-PK1细胞中研究THC对FK506诱导的肾细胞损伤的影响。LLC-PK1细胞先用不同浓度的THC预处理2小时，然后加入FK506处理24小时。对LLC-PK1细胞进行FK506和THC处理，并检测细胞活力和谷胱甘肽水平。使用膜联蛋白V/碘化丙啶染色通过流式细胞术检测凋亡细胞数量。通过蛋白质免疫印迹分析测量caspase-9、caspase-3、B细胞淋巴瘤-2（Bcl-2）和Bcl-2相关X蛋白水平，以确定THC对LLC-PK1细胞凋亡的影响。
THC处理可显著改善FK506诱导的LLC-PK1肾细胞损伤。THC通过防止FK506诱导的谷胱甘肽减少来保护LLC-PK1细胞。THC可保护LLC-PK1细胞免受FK506诱导的细胞凋亡。THC处理组细胞凋亡显著减少，Bcl-2水平升高。THC处理组中Bcl-2相关X蛋白、caspase-3和caspase-9水平降低。
这些结果共同提供了治疗证据表明，THC通过抗氧化作用和抑制细胞凋亡改善FK506诱导的肾损伤。
