Department of Urology, Ningbo Urology and Nephrology Hospital, Ningbo University, Ningbo, Zhejiang 315100, P.R. China.
Ningbo Institute of Medical Sciences, Ningbo University, Ningbo, Zhejiang 315100, P.R. China.
Mol Med Rep. 2018 Apr;17(4):6038-6044. doi: 10.3892/mmr.2018.8591. Epub 2018 Feb 12.
This study explored the potential value of curcumin, a natural product, in the protection of CsA‑induced nephrotoxicity. The aim of the present study was to investigate the effects of curcumin on Cyclosporine A (CsA)‑induced renal oxidative stress and determine the potential underlying molecular mechanisms of the renal protective effects of Cur. HK‑2 human renal cells were co‑treated with CsA and various doses of Cur. Cell survival rate was determined by an MTT assay, total cellular protein was collected and oxidative stress in cell homogenates was evaluated by determining the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH‑Px) and catalase (CAT), the levels of malondialdehyde (MDA) and reactive oxygen species (ROS), and total antioxidant capacity. Furthermore, Bcl‑2 and Bcl‑2‑associated X (Bax) protein expression was measured by western blot analysis. In addition, a CsA‑induced nephrotoxicity (CAN) rat model was also established. Renal function was analyzed by measuring creatinine (Crea) and blood urea nitrogen (BUN) in the serum of rats, and histopathological examination was performed on renal tissues using hematoxylin and eosin staining, periodic acid‑Schiff staining and nuclear factor‑κB (NF‑κB) immunostaining. The results demonstrated that treatment of HK‑2 cells with CsA significantly increased ROS and MDA levels, and decreased the activities of SOD, GSH‑Px and CAT, compared with the control group. However, these effects of CsA were dose‑dependently improved by treatment with Cur. In addition, Cur treatment increased Bcl‑2 and decreased Bax protein in HK‑2 cells, compared with cells treated with CsA alone. In the CAN rat model CsA (30 mg/kg) treatment significantly elevated serum Crea levels and BUN, but lowered endogenous Crea clearance rate, compared with the control group. Co‑administration of Cur with CsA significantly reversed the effects of CsA on serum Crea levels, BUN and Crea clearance rate (Ccr). Additionally, Cur alleviated CsA‑induced renal cell injury, as less vacuolar degeneration of glomerular cells was observed compared with the CsA alone group. In conclusion, Cur may increase renal antioxidant capacity and reduce the Bax/Bcl‑2 ratio, subsequently improving CsA‑induced renal failure and renal tubular deformation and cell vacuolization.
本研究探讨了姜黄素（一种天然产物）在保护环孢素 A（CsA）诱导的肾毒性中的潜在价值。本研究旨在探讨姜黄素对环孢素 A（CsA）诱导的肾氧化应激的影响，并确定姜黄素的肾保护作用的潜在分子机制。将 HK-2 人肾细胞与 CsA 和不同剂量的姜黄素共同孵育。通过 MTT 测定法测定细胞存活率，收集细胞总蛋白，并通过测定超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）的活性、丙二醛（MDA）和活性氧（ROS）的水平以及总抗氧化能力来评估细胞匀浆中的氧化应激。此外，通过 Western blot 分析测定 Bcl-2 和 Bcl-2 相关 X（Bax）蛋白的表达。此外，还建立了环孢素 A 诱导的肾毒性（CAN）大鼠模型。通过测定大鼠血清中的肌酐（Crea）和血尿素氮（BUN）来分析肾功能，并通过苏木精和伊红染色、过碘酸-希夫染色和核因子-κB（NF-κB）免疫染色对肾组织进行组织病理学检查。结果表明，与对照组相比，CsA 处理 HK-2 细胞可显著增加 ROS 和 MDA 水平，并降低 SOD、GSH-Px 和 CAT 的活性。然而，Cur 的处理可剂量依赖性地改善这些 CsA 的作用。此外，与单独用 CsA 处理的细胞相比，Cur 处理可增加 HK-2 细胞中的 Bcl-2 并降低 Bax 蛋白。在 CAN 大鼠模型中，与对照组相比，CsA（30mg/kg）处理可显著升高血清 Crea 水平和 BUN，但降低内源性 Crea 清除率。Cur 与 CsA 联合给药可显著逆转 CsA 对血清 Crea 水平、BUN 和 Crea 清除率（Ccr）的影响。此外，Cur 可减轻 CsA 诱导的肾细胞损伤，与单独用 CsA 处理的组相比，肾小球细胞的空泡变性减少。总之，姜黄素可能通过增加肾抗氧化能力和降低 Bax/Bcl-2 比值，从而改善环孢素 A 诱导的肾衰竭和肾小管变形以及细胞空泡化。
