目的 探讨二精方对顺铂诱导小鼠急性肾损伤（AKI）的影响及机制。方法 将50只SPF级C57BL/6J小鼠随机分为对照组、模型组和二精方低、中、高剂量（335、670、1 340 mg·kg_-1）组，每组10只，ig给药，对照组和模型组ig去离子水，持续20 d。除对照组外，其余各组在第17天单次ip顺铂（15 mg·kg_-1）诱导小鼠AKI模型。72 h后取血和肾脏组织，计算肾脏指数；试剂盒法测定血清中肌酐（CRE）、尿素氮（BUN）水平，测定肾脏组织中丙二醛（MDA）水平和超氧化物歧化酶（SOD）、还原型谷胱甘肽（GSH）、过氧化氢酶（CAT）的活力；试剂盒法检测小鼠血清中白细胞介素1β（IL-1β）和肿瘤坏死因子α（TNF-α）水平、胱抑素C（Cys-C）和肾损伤分子-1（KIM-1）含量；苏木精-伊红（HE）染色法观察肾脏组织的损伤程度； Western blotting法检测肾脏组织中KIM-1、中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、Kelch样ECH相关蛋白1（Keap-1）、核因子E2相关因子2（Nrf2）、血红素加氧酶-1（HO-1）、磷酸化的核因子κB p65（p-NF-κB p65）、TNF-α、IL-6蛋白表达。结果 与模型组比较，二精方组小鼠精神活跃、进食增加、体质量下降减缓；肾脏色泽更红润，肾脏指数显著下降（P＜0.05、0.01）；各剂量组小鼠血清CRE水平显著降低（P＜0.01），中、高剂量组BUN水平显著降低（P＜0.01）；各剂量组血清KIM-1和Cys-C水平均显著降低（P＜0.05、0.01、0.001），中、高剂量组血清IL-1β和TNF-α水平显著降低（P＜0.05、0.01、0.001）；中、高剂量组肾脏中MDA含量显著降低（P＜0.05、0.01），各剂量组GSH、CAT活力显著升高（P＜0.05、0.01），高剂量组SOD活力显著升高（P＜0.01）；各剂量组肾组织结构有改善，肾小管损伤评分显著降低（P＜0.01、0.001）；中、高剂量显著下调KIM-1、NGAL、Keap-1、p-NF-κB p65、IL-6、TNF-α蛋白水平，显著上调HO-1、Nrf2的蛋白水平（P＜0.05、0.01、0.001）。结论 二精方对顺铂诱导的小鼠AKI具有显著保护作用，其机制主要通过双重途径实现：一方面激活Keap-1/Nrf2/HO-1通路增强抗氧化能力，清除活性氧（ROS）并修复线粒体损伤；另一方面抑制NF-κB p65磷酸化及促炎因子释放，打破“氧化应激-炎症”恶性循环。

Objective To investigate the effect and mechanism of Erjing Formula on cisplatin-induced acute kidney injury (AKI) in mice. Methods Fifty SPF-grade C57BL/6J mice were randomly divided into a control group, a model group, and low-, medium-, and high-dose Erjing Formula groups (335, 670, and 1 340 mg·kg^-1, respectively), with 10 mice in each group. The mice were administered ig for 20 d, and the control and model groups were given deionized water. On the 17th day, the model group and the Erjing Formula groups were intraperitoneally (ip) injected with cisplatin (15 mg·kg^-1) to induce AKI. After 72 h, blood and kidney tissues were collected. The kidney index was calculated. The levels of creatinine (CRE) and blood urea nitrogen (BUN) in serum were determined by kits. The levels of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), reduced glutathione (GSH), and catalase (CAT) in kidney tissues were measured. The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in serum, cystatin C (Cys-C), and kidney injury molecule-1 (KIM-1) in serum were detected by kits. The degree of kidney tissue damage was observed by hematoxylin-eosin (HE) staining. The protein expressions of KIM-1, neutrophil gelatinase-associated lipocalin (NGAL), Kelch-like ECH-associated protein 1 (Keap-1), nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase- 1 (HO-1), phosphorylated nuclear factor κB p65 (p-NF-κB p65), TNF-α, and IL-6 in kidney tissues were detected by Western blotting. Results Compared with the model group, the mice in the Erjing Formula groups were more active, had increased food intake, and a slower decrease in body weight. The kidneys were more red and the kidney index was significantly decreased (P < 0.05, 0.01). The serum CRE levels in all dose groups were significantly decreased (P < 0.01), and the BUN levels in the medium- and high-dose groups were significantly decreased (P < 0.01). The serum KIM-1 and Cys-C levels in all dose groups were significantly decreased (P < 0.05, 0.01, 0.001), and the serum IL-1β and TNF-α levels in the medium- and high-dose groups were significantly decreased (P < 0.05, 0.01, 0.001). The MDA content in the kidneys of the medium- and high-dose groups was significantly decreased (P < 0.05, 0.01), and the activities of GSH and CAT in all dose groups were significantly decreased (P < 0.05, 0.01), while the SOD activity in the high-dose group was significantly decreased (P < 0.01). The structure of kidney tissues in all dose groups was improved, and the renal tubular injury score was significantly decreased (P < 0.01, 0.001). The medium- and high-dose groups significantly downregulated the protein levels of KIM-1, NGAL, Keap-1, p-NF-κB p65, IL-6, and TNF-α, and significantly upregulated the protein levels of HO-1 and Nrf2 (P < 0.05, 0.01, 0.001). Conclusion Erjing Formula has a significant protective effect on cisplatin-induced AKI in mice, and its mechanism is mainly achieved through dual pathways: on the one hand, it activates the Keap-1/Nrf2/HO-1 pathway to enhance antioxidant capacity, clear reactive oxygen species (ROS), and repair mitochondrial damage; on the other hand, it inhibits the phosphorylation of NF-κB p65 and the release of pro-inflammatory factors, breaking the "oxidative stress-inflammation" vicious cycle.

R285.5

贵州省科技计划项目[黔科合基础-ZK（2021）一般373]