Pandey Vivek, Sharma Alka, Tiwari Sonal, Patel Yashvant, Chauhan Jayhind Kumar, Ayesha Safiya, Sahu Alakh N, Gupta Rashmi, Tripathi Anima, Dubey Pawan K
Centre for Genetic Disorders, Institute of Science, Banaras Hindu University, Varanasi 221005, India.
Department of Zoology, MMV, Banaras Hindu University, Varanasi 221005, India.
Reprod Toxicol. 2024 Dec;130:108737. doi: 10.1016/j.reprotox.2024.108737. Epub 2024 Oct 26.
Studies provide notable evidence that oxidative stress (OS) mediated reactive oxygen species (ROS) disturb reproductive health. We have shown in our previous publication that exposure of Di-(2-ethylhexyl) phthalate (DEHP), induces OS mediated ROS generation which inhibits steroid synthesis. In the present study, we demonstrated the ameliorative/protective effects of one of the steroidal saponins, i.e., Shatavarin-IV, isolated from the roots of Asparagus racemosus against DEHP induced OS in rat granulosa cells. Granulosa cells were exposed with DEHP alone (400 μM), Shatavarin-IV alone (8 μg/ml), and a combination of DEHP + Shatavarin-IV (400 μM + 8 μg/ml) in vitro for 24 hrs. Intracellular ROS, OS/hypoxia, mitochondrial membrane potential, steroid-responsive genes expression were analyzed. The results revealed that the effective dose of DEHP (400 µg) significantly increased OS compared to the control by increasing ROS levels, mitochondrial membrane potential, and β-galactosidase activity with a higher level of apoptotic genes (Bax, Caspase-3) expression at mRNA level. Further, DEHP significantly (p < 0.05) reduced mRNA expression of steroidogenic responsive genes (StAR, CYP17A1, and CYP19A1) in granulosa cells treated with above combination compared to control. Interestingly, co-treatment of DEHP + Shatavarin-IV significantly suppressed the DEHP induced OS, ROS, β-galactosidase levels and enhanced steroidogeneic and apoptotic gene expression activities, which suggests that Shatavarin-IV rescued DEHP-induced changes that may useful for the prevention of DEHP- induced reproductive toxicity.
研究提供了显著证据，表明氧化应激（OS）介导的活性氧（ROS）会干扰生殖健康。我们在之前的出版物中表明，邻苯二甲酸二（2-乙基己基）酯（DEHP）的暴露会诱导OS介导的ROS生成，从而抑制类固醇合成。在本研究中，我们证明了从印度天门冬根部分离出的一种甾体皂苷—— Shatavarin-IV对DEHP诱导的大鼠颗粒细胞氧化应激具有改善/保护作用。将颗粒细胞在体外分别单独暴露于DEHP（400μM）、Shatavarin-IV（8μg/ml）以及DEHP + Shatavarin-IV（400μM + 8μg/ml）组合中24小时。分析细胞内ROS、OS/缺氧、线粒体膜电位、类固醇反应基因表达。结果显示，与对照组相比，有效剂量的DEHP（400μg）通过提高ROS水平、线粒体膜电位和β-半乳糖苷酶活性，显著增加了氧化应激，且在mRNA水平上凋亡基因（Bax、Caspase-3）表达水平更高。此外，与对照组相比，上述组合处理的颗粒细胞中，DEHP显著（p < 0.05）降低了类固醇生成反应基因（StAR、CYP17A1和CYP19A1）的mRNA表达。有趣的是，DEHP与Shatavarin-IV联合处理显著抑制了DEHP诱导的氧化应激、ROS、β-半乳糖苷酶水平，并增强了类固醇生成和凋亡基因表达活性，这表明Shatavarin-IV挽救了DEHP诱导的变化，这可能有助于预防DEHP诱导的生殖毒性。
