Department of Nephrology, Shenzhen Traditional Chinese Medicine Hospital, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, No.1, Fuhua Road, Futian District, Shenzhen, Guangdong, 518033, China.
BMC Complement Med Ther. 2024 Feb 24;24(1):101. doi: 10.1186/s12906-024-04366-9.
Acute kidney injury (AKI), characterized by necroptosis and activation of MAPK pathway, causes sudden declines in renal function. To date, efficacious treatments are lacking. JianPiYiShen Formula (JPYSF) has a protective effect on the kidneys. The aim of this study is to explore the mechanism of JPYSF in cisplatin-induced AKI.
Male C57/BL6J mice were divided into control group, cisplatin group and cisplatin + JPYSF group. Before establishing the model, the cisplatin + JPYSF group was administered JPYSF (18.35 g/kg/day) by gavage for 5 consecutive days. A single intraperitoneal injection of cisplatin (20 mg/kg) was used to establish AKI model. Measurement of renal function and H&E staining were performed to assess renal damage. WB, PCR, TUNEL staining and immunohistochemistry were used to detect related indicators of mitochondrial function, oxidative stress, necroptosis, inflammation and MAPK pathway. And one-way analysis of variance was used to compare group differences.
Compared with the cisplatin group, JPYSF can attenuate AKI, reflected by the decrease in Scr and BUN levels, the improvement of renal tubular injury, and the downregulation of NGAL and KIM1. Cisplatin can induce mitochondrial dysfunction and oxidative stress, triggering necroptosis. In this study, JPYSF improved mitochondrial dysfunction to enhance oxidative stress, as manifested by upregulation of OPA1, PGC-1α, SOD and CAT, and downregulation of DRP1 and MFF. Then JPYSF showed a significant protective effect in necroptosis, as embodied by reduced number of TUNEL-positive cells, decreased the gene expression of RIPK3 and MLKL, as well as downregulation the proteins expression of P-RIPK1, P-RIPK3, and P-MLKL. Moreover, necroptosis can aggravate inflammation. JPYSF ameliorated inflammation by improving inflammatory and anti-inflammatory indexes, including downregulation of TNF-α, IL-6, MCP-1 and LY6G, and upregulation of IL-10. In addition, JPYSF also inhibited MAPK pathway to improve necroptosis by decreasing the expression of P-JNK and P-ERK.
Our data showed that JPYSF prevents cisplatin-induced AKI by improving necroptosis through MAPK pathway, which is related to the improvement of mitochondrial dysfunction, oxidative stress, and inflammation.
急性肾损伤（AKI）的特征是坏死和 MAPK 通路的激活，导致肾功能突然下降。迄今为止，缺乏有效的治疗方法。健脾益肾方（JPYSF）对肾脏有保护作用。本研究旨在探讨 JPYSF 对顺铂诱导的 AKI 的作用机制。
雄性 C57/BL6J 小鼠分为对照组、顺铂组和顺铂+JPYSF 组。在建立模型之前，顺铂+JPYSF 组连续 5 天通过灌胃给予 JPYSF（18.35 g/kg/天）。单次腹腔注射顺铂（20 mg/kg）建立 AKI 模型。测定肾功能和 H&E 染色评估肾损伤。采用 WB、PCR、TUNEL 染色和免疫组化检测线粒体功能、氧化应激、坏死、炎症和 MAPK 通路相关指标。采用单因素方差分析比较组间差异。
与顺铂组相比，JPYSF 可减轻 AKI，表现为 Scr 和 BUN 水平降低，肾小管损伤改善，NGAL 和 KIM1 下调。顺铂可诱导线粒体功能障碍和氧化应激，引发坏死。在本研究中，JPYSF 通过上调 OPA1、PGC-1α、SOD 和 CAT，下调 DRP1 和 MFF，改善线粒体功能，增强氧化应激，从而改善线粒体功能障碍。然后，JPYSF 通过减少 TUNEL 阳性细胞数、降低 RIPK3 和 MLKL 基因表达以及下调 P-RIPK1、P-RIPK3 和 P-MLKL 蛋白表达，在坏死中表现出显著的保护作用。此外，坏死可加重炎症。JPYSF 通过改善 TNF-α、IL-6、MCP-1 和 LY6G 下调和 IL-10 上调，改善炎症和抗炎指标，从而改善炎症。此外，JPYSF 还通过降低 P-JNK 和 P-ERK 的表达，抑制 MAPK 通路，改善坏死。
我们的数据表明，JPYSF 通过 MAPK 通路改善坏死来预防顺铂诱导的 AKI，这与改善线粒体功能障碍、氧化应激和炎症有关。
