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客户采用我司G-NH2氨基磁珠偶联黄芩素钓取作用蛋白在Journal of Hazardous Materials发表高分论文

2023-12-5 20:58:19点击:

Na Sun, et al. Scutellarin targets Wnt5a against zearalenone-induced apoptosis in mouse granulosa cells in vitro and in vivo. Journal of Hazardous Materials, Volume 464, 15 February 2024, 132917. https://doi.org/10.1016/j.jhazmat.2023.132917  (G-NH2磁珠偶联羧基化合物)


Scutellarin targets Wnt5a against zearalenone-induced apoptosis in mouse granulosa cells in vitro and in vivo


Abstract
Zearalenone (ZEA) poses severe reproductive toxicity to both humans and animals. Scutellarin has been demonstrated to rescue ZEA-induced apoptosis in mouse ovarian granulosa cells (GCs), but its specific targets remain unclear. In the present study, the potential targets of scutellarin were determined to clarify the mechanisms of scutellarin against ZEA-induced ovarian damage. 287 targets of scutellarin in mouse ovarian GCs were obtained by magnetic nano-probe-based fishing assay and liquid chromatography-tandem mass spectrometry. Wnt5a had the lowest binding free energy with scutellarin at − 8.3 kcal/mol. QRT-PCR and western blot showed that scutellarin significantly increased the Wnt5a and β-catenin expression compared with the ZEA-treated group, and cleaved-caspase-3 expression was significantly increased in the scutellarin-treated group after interfering with the expression of Wnt5a. The affinity constant (KD) of Wnt5a and scutellarin was 1.7 × 10−5 M. The pull-down assay also demonstrated that scutellarin could specifically bind to Wnt5a protein. Molecular docking results showed that scutellarin could form hydrogen bonds with TRY52, GLN56, and SER90 on Wnt5a protein, and western blot assay confirmed SER90 was an important site for the binding. Scutellarin significantly increased Wnt5a and β-catenin expression and decreased cleaved-caspase-3 expression in ovarian tissues of mice. In conclusion, scutellarin exerted anti-apoptotic effects on ZEA-induced mouse ovarian GCs by targeting Wnt5a.


摘要
玉米赤霉烯酮(ZEA)对人类和动物都具有严重的生殖毒性。黄芩素已被证明可以挽救 ZEA 诱导的小鼠卵巢颗粒细胞 (GC) 细胞凋亡,但其具体靶点尚不清楚。本研究确定了黄芩素的潜在靶点,以阐明黄芩素对ZEA诱导的卵巢损伤的机制。采用磁性纳米探针法和液相色谱-串联质谱法,获得小鼠卵巢GCs中黄芩素的287个靶标。Wnt5a与黄芩素的结合自由能最低,为−8.3 kcal/mol,QRT-PCR和Western blot结果显示,黄芩素与ZEA处理组相比,黄芩蛋白显著增加Wnt5a和β-catenin表达,黄芩素处理组在干扰Wnt5a表达后,裂解半胱天冬酶-3表达显著升高。Wnt5a和黄芩素的亲和常数(KD)为1.7 × 10−5 M。pull-down 试验还表明,黄芩素可以特异性地与 Wnt5a 蛋白结合。分子对接结果显示,黄芩素可与TRY52、GLN56和SER90在Wnt5a蛋白上形成氢键,Western blot试验证实SER90是该蛋白结合的重要位点。黄芩素显著增加小鼠卵巢组织中Wnt5a和β-catenin的表达,降低cleaved-caspase-3的表达。综上所述,黄芩素通过靶向Wnt5a对ZEA诱导的小鼠卵巢GCs发挥抗凋亡作用。


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