Singh Anubha, Tandon Simran, Nandi Shoma Paul, Kaur Tanzeer, Tandon Chanderdeep
Amity Institute of Biotechnology, Amity University, Noida, India.
Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Noida, India.
J Ethnopharmacol. 2021 Jul 15;275:114104. doi: 10.1016/j.jep.2021.114104. Epub 2021 Apr 6.
In the Indian traditional system of medicine, Bergenia ligulata (Wall.) Engl. has been used for treatment of urolithiasis. Its efficacious nature has led to its incorporation in various commercial herbal formulations such as Cystone and Neeri which are prescribed for kidney related ailments.
To assess whether ethanolic extract of B. ligulata can mitigate the cascade of inflammatory responses that cause oxidative stress and ultimately cell death in renal epithelial cells exposed to hyperoxaluric conditions.
Bioactivity guided fractionation using solvents of varying polarities was employed to evaluate the potential of the extracts of B. ligulata to inhibit the crystallization process. Modulation of crystal morphology was visualized through Scanning electron microscopy (SEM) analysis. Cell death was assessed using flow cytometry based assays. Alteration in the inflammatory mediators was evaluated using real time PCR and immunocytochemistry. Phytochemical characterization of the ethanolic extract was carried out using FTIR, LC-MS and GC-MS.
Bioactivity guided fractionation for the assessment of antilithiatic activity revealed dose dependent inhibition of nucleation and aggregation process of calcium oxalate crystals in the presence of various extracts, however ethanolic extract showed maximum inhibition and was chosen for further experiments. Studies on renal epithelial NRK-52E cells showed, cytoprotective efficacy of B. ligulata extract against oxalate injury. SEM anaysis further revealed the potential of the extract to modulate the crystal structure and adhesion to renal cell surface. Exposure of the renal cells to the extract led to conversion of the calcium oxalate monohydrate (COM) crystals to the less injurious calcium oxalate dihydrate (COD) form. Expression analysis for oxidative stress and inflammatory biomarkers in NRK-52E cells revealed up-regulation of Mitogen activated protein kinase (MAPK), Osteopontin (OPN) and Nuclear factor- ĸB (NF-ĸB), in response to calcium oxalate insult; which was drastically reduced in the presence of B. ligulata extract. Flow cytometric evaluation pointed to caspase 3 mediated apoptotic cell death in oxalate injured cells, which was attenuated by B. ligulata extract.
Considering the complex multifactorial etiology of urolithiasis, ethanolic extract from B. ligulata can be a promising option for the management of kidney stones, as it has the potential to limit inflammation and the subsequent cell death.
在印度传统医学体系中，岩白菜（Bergenia ligulata (Wall.) Engl.）被用于治疗尿石症。其显著疗效使其被纳入各种商业草药配方中，如用于治疗肾脏相关疾病的Cystone和Neeri。
评估岩白菜乙醇提取物是否能够减轻在高草酸环境下肾上皮细胞中引发氧化应激并最终导致细胞死亡的一系列炎症反应。
采用不同极性溶剂进行生物活性导向分级分离，以评估岩白菜提取物抑制结晶过程的潜力。通过扫描电子显微镜（SEM）分析观察晶体形态的变化。使用基于流式细胞术的检测方法评估细胞死亡情况。利用实时聚合酶链反应（PCR）和免疫细胞化学技术评估炎症介质的变化。使用傅里叶变换红外光谱（FTIR）、液相色谱 - 质谱联用（LC - MS）和气相色谱 - 质谱联用（GC - MS）对乙醇提取物进行植物化学特征分析。
用于评估抗结石活性的生物活性导向分级分离显示，在各种提取物存在的情况下，草酸钙晶体的成核和聚集过程呈现剂量依赖性抑制，然而乙醇提取物显示出最大抑制作用，并被选用于进一步实验。对肾上皮NRK - 52E细胞的研究表明，岩白菜提取物对草酸损伤具有细胞保护作用。SEM分析进一步揭示了该提取物调节晶体结构以及与肾细胞表面粘附的潜力。将肾细胞暴露于该提取物导致一水草酸钙（COM）晶体转变为危害性较小的二水草酸钙（COD）形式。对NRK - 52E细胞中氧化应激和炎症生物标志物的表达分析显示，在草酸刺激下，丝裂原活化蛋白激酶（MAPK）、骨桥蛋白（OPN）和核因子 - κB（NF - κB）上调；而在岩白菜提取物存在的情况下，这种上调显著降低。流式细胞术评估表明，草酸损伤细胞中存在半胱天冬酶3介导白勺凋亡细胞死亡，而岩白菜提取物可减轻这种死亡。
考虑到尿石症复杂的多因素病因，岩白菜乙醇提取物可能是治疗肾结石的一个有前景的选择，因为它有潜力限制炎症及随后的细胞死亡。
