Kaushik Jyoti, Tandon Simran, Gupta Varun, Nayyar Jasamrit, Singla Surinder Kumar, Tandon Chanderdeep
Amity Institute of Biotechnology, Amity University, Noida, India.
Amity Institute of Molecular Medicine & Stem Cell Research, Amity University, Noida, India.
PLoS One. 2017 Aug 28;12(8):e0183218. doi: 10.1371/journal.pone.0183218. eCollection 2017.
Tribulus terrestris has significant antilithiatic efficacy established via both in vitro as well as in vivo studies and is used in numerous anti-urolithiatic herbal formulations viz. Cystone, Uriflow, Uritone and Neeri. However, to fully utilize its antilithiatic potential, the influence of different extraction parameters on antilithiatic ability of T. terrestris aqueous extract needs elucidation. Thus, the current study was undertaken using statistically optimized extraction conditions for aqueous extract preparation. Response surface methodology was employed to observe the influence of three variables i.e. temperature (°C), time (h) and solid: liquid ratio (S: L) on the extraction yield (%) and protein content (mg/g) of T. terrestris aqueous extract. RSM results revealed that the high S:L ratio, low temperature and reduced incubation time were optimal conditions for aqueous extraction. Under such extraction conditions the protein content reached the value of 26.6±1.22 mg/g and the obtained extraction yield was 27.32±1.62%. The assessment of antilithiatic activity of 4 selected extracts (AE1-4), revealed enhanced nucleation and aggregation inhibition of calcium oxalate crystals with AE1 and AE2, which in addition significantly altered the size and morphology of calcium oxalate monohydrate (COM) crystals compared to AE3 and AE4. In vitro cell culture based studies on renal epithelial cells (MDCK, NRK-52E and PK 15) proved that the AE1 showed higher cytoprotective potency by increasing cell viability as compared to the oxalate treated group. The free radical scavenging activity of aqueous extract lowered the reactive oxygen specie's induced damage and potentially reduced the signals of programmed cell death due to oxalate injury. In addition, modulation of the COM crystal morphology was enhanced by AE1 as compared to AE2. The FTIR and GC-MS analysis of AE1, showed the presence of biomolecules which could aid in the attenuation of lithiatic process. In the light of these results the utility of the RSM approach to fully optimize the antilithiatic potential of T. terrestris cannot be undermined.
刺蒺藜经体外和体内研究已证实具有显著的抗结石功效，并被用于多种抗尿路结石的草药配方中，如消石素、尿石通、尿通宁和尼里。然而，为了充分发挥其抗结石潜力，需要阐明不同提取参数对刺蒺藜水提取物抗结石能力的影响。因此，本研究采用统计优化的提取条件制备水提取物。采用响应面法观察温度（℃）、时间（h）和固液比（S:L）这三个变量对刺蒺藜水提取物提取率（%）和蛋白质含量（mg/g）的影响。响应面法结果表明，高S:L比、低温和缩短孵育时间是水提取的最佳条件。在这种提取条件下，蛋白质含量达到26.6±1.22 mg/g，提取率为27.32±1.62%。对4种选定提取物（AE1-4）的抗结石活性评估显示，AE1和AE2增强了草酸钙晶体的成核和聚集抑制作用，与AE3和AE4相比，它们还显著改变了一水合草酸钙（COM）晶体的大小和形态。基于肾上皮细胞（MDCK、NRK-52E和PK 15）的体外细胞培养研究证明，与草酸盐处理组相比，AE1通过提高细胞活力表现出更高的细胞保护效力。水提取物的自由基清除活性降低了活性氧诱导的损伤，并可能减少了草酸盐损伤导致的程序性细胞死亡信号。此外，与AE2相比，AE1增强了对COM晶体形态的调节。AE1的傅里叶变换红外光谱（FTIR）和气相色谱-质谱联用（GC-MS）分析表明，存在有助于减轻结石形成过程的生物分子。鉴于这些结果，响应面法在充分优化刺蒺藜抗结石潜力方面的实用性不可忽视。
