School of Pharmaceutical Science and Technology, Faculty of Medicine, Tianjin University, Tianjin 300072, China.
School of Medicine, Nankai University, Tianjin 300071, China.
Phytomedicine. 2024 Dec;135:156145. doi: 10.1016/j.phymed.2024.156145. Epub 2024 Oct 11.
The recurrent nature and socioeconomic burden of nephrolithiasis demand effective treatments. Delineating the crosstalk between inflammatory processes and the endogenous oxalate metabolism pathway, which underpins nephrolithiasis pathogenesis, is essential for advancing treatment strategies.
We aim to screen therapeutic Chinese herbal remedies and their bioactive constituents for kidney stone treatment using a fruit fly model, followed by efficacy and mechanism validation in a rodent nephrolithiasis model as well as in vitro human cell culture model.
We developed a fruit fly model to screen for efficient traditional Chinese medicine herbs and their active compounds for kidney stone treatment. Candidate active compounds from efficient herbs were separated and identified by solid-phase chromatography coupled with LC-MS analysis. Fruit fly genetic tools were employed to manipulate the expression of related genes to explore the therapeutic mechanisms of the Lycii Cortex and kukoamine A (KuA). To confirm the therapeutic effects and mechanisms of KuA for mammalian nephrolithiasis, mouse model of glyoxylate-induced kidney stone and human renal tubular cells were used. The therapeutic role of kukoamine A in nephrolithiasis was evaluated by assessing tubular injury, crystal deposition, and adhesion. The level of expression and phosphorylation in cells and mice was assessed using RT-qPCR and western blot.
Our findings indicate that Lycii Cortex potently inhibits calcium oxalate stone formation via activation of the JNK/Upd3/JAK/STAT signaling cascade, resulting in diminished endogenous oxalate synthesis by downregulating D-amino acid oxidase (DAO) gene expression, predominantly in fruit fly Malpighian tube stellate cells. KuA was identified as the principal bioactive constituent mediating these effects. Both mouse models and human cell assays confirmed KuA's efficacy in preventing calcium oxalate nephrolithiasis in mammals, through hepatic JAK/STAT3 pathway activation and upregulation of IL-6, culminating in reduced urinary crystal deposition.
Our research underscores the potential of kukoamine A as a lead compound in treating nephrolithiasis and reveals the interplay between the IL-6/JAK/STAT3 inflammatory pathway and endogenous oxalate metabolism in nephrolithiasis pathogenesis. Additionally, it highlights the utility of the fruit fly model as a powerful tool for deciphering the therapeutic mechanisms of traditional Chinese herbs.
肾结石的反复发作和巨大的社会经济负担需要有效的治疗方法。阐明炎症过程与内源性草酸代谢途径之间的相互作用，对于推进肾结石发病机制的治疗策略至关重要。
我们旨在使用果蝇模型筛选治疗肾结石的中药及其生物活性成分，并在肾结石的啮齿动物模型以及体外人细胞培养模型中验证其疗效和机制。
我们开发了一种果蝇模型，用于筛选治疗肾结石的有效中药及其活性化合物。通过固相色谱与 LC-MS 分析分离和鉴定有效草药中的候选活性化合物。使用果蝇遗传工具来操纵相关基因的表达，以探索枸杞皮和苦柯胺 A（KuA）的治疗机制。为了确认 KuA 对哺乳动物肾结石的治疗效果和机制，使用了乙醛酸盐诱导的小鼠肾结石模型和人肾小管细胞。通过评估肾小管损伤、晶体沉积和黏附，来评估 KuA 在肾结石中的治疗作用。使用 RT-qPCR 和 Western blot 评估细胞和小鼠中表达和磷酸化水平。
我们的研究结果表明，枸杞皮通过激活 JNK/Upd3/JAK/STAT 信号级联反应，强烈抑制草酸钙结石的形成，从而通过下调 D-氨基酸氧化酶（DAO）基因的表达，减少内源性草酸的合成，主要在果蝇的马尔皮基氏管星状细胞中。鉴定出 KuA 是介导这些作用的主要生物活性成分。小鼠模型和人细胞实验均证实 KuA 可通过激活肝脏 JAK/STAT3 通路和上调白细胞介素 6（IL-6）来预防哺乳动物的草酸钙肾结石，从而减少尿晶体沉积。
本研究强调了苦柯胺 A 作为治疗肾结石的潜在先导化合物的重要性，并揭示了 IL-6/JAK/STAT3 炎症通路与肾结石发病机制中内源性草酸代谢之间的相互作用。此外，它还突出了果蝇模型作为破译中药治疗机制的有力工具的实用性。
