Gaikwad Anil Bhanudas, Hajare Aditya Dipakrao, Kulkarni Hrushikesh, Bhadange Rohan
Department of Pharmacy, Birla Institute of Technology and Science, Pilani, Pilani Campus, Vidya Vihar, Pilani, Rajasthan 333031, India.
Department of Pharmacy, Birla Institute of Technology and Science, Pilani, Pilani Campus, Vidya Vihar, Pilani, Rajasthan 333031, India.
Biochem Biophys Res Commun. 2025 May 26;760:151711. doi: 10.1016/j.bbrc.2025.151711. Epub 2025 Mar 26.
Diabetic kidney disease (DKD) remains a leading cause of end-stage kidney disease worldwide, necessitating novel therapeutic approaches. This study investigated the renoprotective potential of recombinant human α-Klotho (α-Klotho; KL) as an adjunctive therapy to telmisartan (TEL) in streptozotocin-induced DKD rats and high glucose and exogenous TGF-β1 (rh-TGF-β1) -exposed NRK-52E cells. The combination therapy significantly preserved kidney function parameters, including reduced blood urea nitrogen, serum creatinine, and kidney weight/body weight ratio compared to monotherapy or untreated DKD groups. Histopathological analyses revealed that the combination therapy markedly ameliorated glomerular hypertrophy, tubular atrophy, and interstitial fibrosis as evidenced by H&E, PAS, and Masson's trichrome staining. Immunohistochemical evaluation demonstrated significant downregulation of AT1R and TGF-β1 expression in the combination group. In-vitro studies confirmed that co-treatment enhanced cell viability and preserved normal epithelial morphology of NRK-52E cells under high glucose conditions. Mechanistically, the combination therapy modulated the apoptotic pathway by upregulating Bcl-2 and downregulating Bax expression while simultaneously reducing phosphorylated NF-κB levels, suggesting suppression of inflammation-mediated tubular apoptosis. Additionally, the combination therapy effectively reduced extracellular matrix (ECM) deposition by decreasing fibronectin levels (ELISA) and collagen expressions (RT-PCR). Immunofluorescence analysis demonstrated reduced vimentin expression, indicating a lower degree of epithelial-mesenchymal transition (EMT), along with an increase in α-SMA levels in the animal tissues. These findings demonstrate that adjunctive α-Klotho therapy with telmisartan exerts synergistic renoprotective effects through anti-fibrotic, anti-inflammatory, and anti-apoptotic mechanisms. This combination approach represents a promising therapeutic strategy for DKD management that warrants further clinical investigation.
糖尿病肾病（DKD）仍是全球终末期肾病的主要病因，因此需要新的治疗方法。本研究探讨了重组人α-klotho（α-Klotho；KL）作为替米沙坦（TEL）辅助治疗药物，对链脲佐菌素诱导的DKD大鼠以及高糖和外源性转化生长因子-β1（rh-TGF-β1）处理的NRK-52E细胞的肾脏保护作用。与单一疗法或未治疗的DKD组相比，联合治疗显著保留了肾功能参数，包括降低的血尿素氮、血清肌酐以及肾重/体重比。组织病理学分析显示，联合治疗显著改善了肾小球肥大、肾小管萎缩和间质纤维化，苏木精-伊红染色（H&E）、过碘酸-雪夫染色（PAS）和马松三色染色均证实了这一点。免疫组化评估显示联合组中血管紧张素Ⅱ1型受体（AT1R）和转化生长因子-β1表达显著下调。体外研究证实，联合处理可提高细胞活力，并在高糖条件下维持NRK-52E细胞的正常上皮形态。从机制上讲，联合治疗通过上调Bcl-2和下调Bax表达来调节凋亡途径，同时降低磷酸化核因子-κB水平，提示抑制炎症介导的肾小管凋亡。此外，联合治疗通过降低纤连蛋白水平（酶联免疫吸附测定）和胶原蛋白表达（逆转录-聚合酶链反应）有效减少了细胞外基质（ECM）沉积。免疫荧光分析显示波形蛋白表达降低，表明上皮-间质转化（EMT）程度较低，同时动物组织中α-平滑肌肌动蛋白（α-SMA）水平升高。这些发现表明，α-Klotho与替米沙坦联合治疗通过抗纤维化、抗炎和抗凋亡机制发挥协同肾脏保护作用。这种联合方法代表了一种有前景的DKD治疗策略，值得进一步的临床研究。
