Department of Pharmacy, The Second Xiangya Hospital of Central South University, Changsha, 410011, China; Hunan Provincial Engineering Research Center of Translational Medical and Innovative Drug, The Second Xiangya Hospital of Central South University, Changsha, Hunan, 410011, China.
Department of Geriatrics, The Second Xiangya Hospital of Central South University, Changsha, Hunan, 410011, China.
Biochem Biophys Res Commun. 2021 Sep 10;569:132-138. doi: 10.1016/j.bbrc.2021.06.097. Epub 2021 Jul 7.
Cisplatin-induced acute kidney injury (AKI) is associated with high morbidity and mortality worldwide, but the underlying mechanisms are not fully understood. Downstream-of-kinase 3 (Dok3), a member of the Dok family of adaptor proteins plays a critical role in inflammatory response and immune regulation; however, the role of Dok3 in cisplatin-induced AKI remains unclear. This study explored the effect and potential molecular mechanisms of Dok3 in cisplatin-induced AKI using Dok3 knockout (Dok3) and control mice (129S) with or without administration of a single intraperitoneal injection of cisplatin. Apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, lactate dehydrogenase (LDH) release, and Hoechst staining. Inflammatory factors were measured using ELISA kits. Protein and gene expression levels were measured by western blot analysis and real-time PCR, respectively. The results showed that Dok3 was expressed in renal tubular epithelial cells. Dok3 expression was decreased in kidneys of mice treated with cisplatin and cisplatin-treated HK2 cells. Dok3 mice showed lower creatinine levels and NGAL expression, and increased survival rates compared to 129S mice. Cisplatin-induced production of TNF-α and IL-6, and renal tubular cell apoptosis was attenuated in Dok3 mice. In vitro experiments demonstrated that HK2 cells overexpressing Dok3 exhibited exacerbated cisplatin-induced apoptosis and production of TNF-α and IL-6. These findings demonstrate that Dok3 regulates cisplatin-induced AKI by regulating apoptosis and inflammation.
顺铂诱导的急性肾损伤（AKI）在全球范围内与高发病率和死亡率相关，但潜在机制尚未完全阐明。激酶 3 下游（Dok3）是衔接蛋白家族的一个成员，在炎症反应和免疫调节中起着关键作用；然而，Dok3 在顺铂诱导的 AKI 中的作用尚不清楚。本研究使用 Dok3 敲除（Dok3）和对照（129S）小鼠，或在单次腹腔注射顺铂后，探讨了 Dok3 在顺铂诱导的 AKI 中的作用及其潜在的分子机制。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）检测、乳酸脱氢酶（LDH）释放和 Hoechst 染色评估细胞凋亡。通过 ELISA 试剂盒检测炎症因子。通过 Western blot 分析和实时 PCR 分别测量蛋白和基因表达水平。结果表明，Dok3 在肾小管上皮细胞中表达。顺铂处理的小鼠肾脏中 Dok3 表达减少，顺铂处理的 HK2 细胞中 Dok3 表达减少。与 129S 小鼠相比，Dok3 小鼠的肌酐水平和 NGAL 表达降低，存活率增加。Dok3 小鼠中顺铂诱导的 TNF-α和 IL-6 产生以及肾小管细胞凋亡减少。体外实验表明，过表达 Dok3 的 HK2 细胞中顺铂诱导的细胞凋亡和 TNF-α和 IL-6 的产生加剧。这些发现表明，Dok3 通过调节细胞凋亡和炎症来调节顺铂诱导的 AKI。
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