Singh Anubha, Kumari Kshama, Tandon Simran, Kaur Tanzeer, Tandon Chanderdeep
School of Biotechnology, Gautam Buddha University, Greater Noida, Uttar Pradesh, India.
Amity University Uttar Pradesh, Noida, India.
Urolithiasis. 2025 Jun 30;53(1):131. doi: 10.1007/s00240-025-01794-1.
Oxidative stress is a key driver of inflammatory injury which predisposes renal cells to nephrolithiatic damage. Recurrent stone formation is associated with cellular dysfunction, oxidative stress, inflammation and renal epithelial cell death. Didymocarpus pedicellata is an important medicinal herb in the ayurvedic system for the treatment of renal afflictions. The phytochemicals present in medicinal plants like D. pedicellata show potential in mitigating these series of cellular responses. Bioactivity guided fractionation, crystallization assays, phytochemical screening and GCMS analysis were performed to assess secondary metabolites present in D. pedicellata. The cytoprotective potential of the extract was evaluated by assessing cell viability, reactive oxygen species (ROS) generation, analyzing cell death, and gene expression changes to determine the extent of damage caused by oxalate exposure. The crystal-cell interface, adhesion dynamics, and morphological alterations were characterized using histological analysis via hematoxylin and eosin staining and ultrastructural examination by scanning electron microscopy. Hyperoxaluric rat model was generated by treating the rats with ethylene glycol and ammonium chloride to induce nephrocalcinosis and the effects of treatment with D. pedicellata were evaluated using renal physiology biomarkers, microscopic analysis of urine and histological studies for crystal deposition and kidney tissue injury. Gene expression analysis of p38 Mitogen-activated protein kinase and Osteopontin was done to assess the expression levels due to hyperoxaluria and treatment with D. pedicellata extract. D. pedicellata extract showed cytoprotective potential through in vitro and in vivo studies. Antilithiatic efficacy was evaluated through crystallization assays which led to metabolic profiling through GCMS analysis exhibiting a wide range of secondary metabolites with antioxidant and anti-inflammatory properties. Our study demonstrated that oxalate injured NRK52-E renal epithelial cells when cotreated with ethanolic extract of D. pedicellata led to an overall increase in cell viability, reduced production of intracellular reactive oxygen species (ROS) which lowered oxidative damage in the cells and decreased apoptosis. Staining the tissues with H&E and SEM imaging revealed modulation of crystal structure and enhanced adhesion to renal epithelial cells. Further evaluation of calcium oxalate deposition in rat model revealed that D. pedicellata alleviated crystal deposition and kidney tissue injury as evaluated by urine analysis and histological studies and decreased expression of inflammatory markers p38MAPK and Osteopontin. This study demonstrates that D. pedicellata exerts cytoprotective effect against calcium oxalate induced nephrocalcinosis by reducing the production of reactive species, oxidative stress, lowering inflammation and apoptosis finally reducing renal injury.
氧化应激是炎症性损伤的关键驱动因素，使肾细胞易患肾结石损伤。复发性结石形成与细胞功能障碍、氧化应激、炎症及肾上皮细胞死亡有关。双距花是阿育吠陀体系中治疗肾脏疾病的重要草药。像双距花这样的药用植物中含有的植物化学物质在减轻这一系列细胞反应方面显示出潜力。进行了生物活性导向分级分离、结晶试验、植物化学筛选和气相色谱-质谱联用分析，以评估双距花中存在的次生代谢产物。通过评估细胞活力、活性氧（ROS）生成、分析细胞死亡以及基因表达变化，来确定草酸盐暴露所造成的损伤程度，从而评估提取物的细胞保护潜力。通过苏木精和伊红染色的组织学分析以及扫描电子显微镜的超微结构检查，对晶体-细胞界面、黏附动力学和形态学改变进行了表征。通过用乙二醇和氯化铵处理大鼠来诱导肾钙质沉着，从而建立高草酸尿大鼠模型，并使用肾脏生理生物标志物、尿液显微镜分析以及晶体沉积和肾组织损伤的组织学研究，来评估双距花治疗的效果。对p38丝裂原活化蛋白激酶和骨桥蛋白进行基因表达分析，以评估高草酸尿症和双距花提取物治疗后的表达水平。通过体外和体内研究，双距花提取物显示出细胞保护潜力。通过结晶试验评估抗结石疗效，该试验通过气相色谱-质谱联用分析进行代谢谱分析，显示出具有抗氧化和抗炎特性的多种次生代谢产物。我们的研究表明，当草酸盐与双距花乙醇提取物共同处理时，草酸盐会损伤NRK52-E肾上皮细胞，而双距花乙醇提取物会使细胞活力总体增加，细胞内活性氧（ROS）生成减少，从而降低细胞内的氧化损伤并减少细胞凋亡。用苏木精和伊红对组织进行染色以及扫描电子显微镜成像显示，晶体结构得到调节，与肾上皮细胞的黏附增强。在大鼠模型中对草酸钙沉积的进一步评估表明，通过尿液分析和组织学研究评估，双距花减轻了晶体沉积和肾组织损伤，并降低了炎症标志物p38丝裂原活化蛋白激酶和骨桥蛋白的表达。这项研究表明，双距花通过减少活性物质的产生、氧化应激、降低炎症和细胞凋亡，最终减轻肾损伤，从而对草酸钙诱导的肾钙质沉着发挥细胞保护作用。
