Acute kidney injury (AKI) is responsible for significant mortality among hospitalized patients that is especially troubling aged people. An effective self-made Chinese medicine formula, Xiaoyu Xiezhuo Drink (XXD), displayed therapeutic effects on AKI. However, the compositions and underlying mechanisms of XXD remain to be elucidated. In this study, we used the ultra-high-performance liquid chromatography method coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) to investigate the chemical components in XXD. Then, the absorbable components of XXD were identified based on the five principles and inputted into the SwissTargetPrediction and STITCH databases to identify the drug targets. AKI-related targets were collected from the GenCLiP 3, GeneCards, and DisGeNET databases. The crossover genes of XXD and AKI were identified for functional enrichment analysis. The protein-protein interaction (PPI) network of crossover genes was constructed, followed by the identification of hub genes. Subsequently, the effects and potential mechanisms of XXD on AKI predicted by the network pharmacology and bioinformatics analyses were experimentally validated in ischemia-reperfusion (I/R) injury-induced AKI aged mouse models. A total of 122 components in XXD were obtained; among them, 58 components were found that could be absorbed in the blood. There were 800 potential drug targets predicted from the 58 absorbable components in AKI which shared 36 crossover genes with AKI-related targets. The results of functional enrichment analysis indicated that crossover genes mostly associated with the response to oxidative stress and the HIF1 signaling pathway. In the PPI network analysis, 12 hub genes were identified, including ALB, IL-6, TNF, TP53, VEGFA, PTGS2, TLR4, NOS3, EGFR, PPARG, HIF1A, and HMOX1. In AKI aged mice, XXD prominently alleviated I/R injury-induced renal dysfunction, abnormal renal pathological changes, and cellular senescence, inflammation, and oxidative damage with a reduction in the expression level of the inflammatory mediator, α-SMA, collagen-1, F4/80, TP53, VEGFA, PTGS2, TLR4, NOS3, EGFR, PPARG, HIF1A, ICAM-1, TGF-β1, Smad3, and p-Smad3 and an increase of nephridial tissue p-H3, Ki67, HMOX1, MMP-9, and Smad7 levels. In summary, our findings suggest that XXD has renoprotective effects against AKI in aged mice via inhibiting the TGF-β1/Smad3 and HIF1 signaling pathways.

中文翻译：

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小鱼泻浊饮通过抑制 TGF-β1/Smad3 和 HIF1 信号通路保护老年小鼠缺血再灌注急性肾损伤

急性肾损伤 (AKI) 是住院患者死亡率显着的原因，尤其是老年人。一种有效的自制中药配方小鱼泻浊饮（XXD）对 AKI 显示出治疗效果。然而，XXD 的组成和潜在机制仍有待阐明。在本研究中，我们使用超高效液相色谱法结合混合三重四极杆飞行时间质谱 (UHPLC-Q-TOF-MS) 来研究 XXD 中的化学成分。然后，根据五个原则确定XXD的可吸收成分并输入SwissTargetPrediction和STITCH数据库以识别药物靶点。AKI 相关目标是从 GenCLiP 3、GeneCards 和 DisGeNET 数据库中收集的。鉴定XXD和AKI的交叉基因用于功能富集分析。构建了交叉基因的蛋白质-蛋白质相互作用（PPI）网络，然后鉴定了枢纽基因。随后，通过网络药理学和生物信息学分析预测的 XXD 对 AKI 的影响和潜在机制在缺血再灌注 (I/R) 损伤诱导的 AKI 老年小鼠模型中得到了实验验证。共获得XXD中的122个组件；其中，58种成分被发现可以被血液吸收。从 AKI 中的 58 种可吸收成分预测了 800 个潜在的药物靶点，这些成分与 AKI 相关靶点共享 36 个交叉基因。功能富集分析结果表明，交叉基因主要与氧化应激反应和HIF1信号通路有关。在 PPI 网络分析中，确定了 12 个中枢基因，包括 ALB、IL-6、TNF、TP53、VEGFA、PTGS2、TLR4、NOS3、EGFR、PPARG、HIF1A 和 HMOX1。在 AKI 老年小鼠中，XXD 显着减轻了 I/R 损伤引起的肾功能障碍、异常肾脏病理变化以及细胞衰老、炎症和氧化损伤，并降低了炎症介质的表达水平，α -SMA、collagen-1、F4/80、TP53、VEGFA、PTGS2、TLR4、NOS3、EGFR、PPARG、HIF1A、ICAM-1、TGF- β1、Smad3和p-Smad3以及肾组织p的增加-H3、Ki67、HMOX1、MMP-9 和 Smad7 水平。总之，我们的研究结果表明，XXD 通过抑制 TGF- β1 /Smad3 和 HIF1 信号通路对老年小鼠的 AKI 具有肾脏保护作用。