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磁珠偶聯(lián)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）對(duì)人類和動(dòng)物都具有嚴(yán)重的生殖毒性。黃芩素已被證明可以挽救 ZEA 誘導(dǎo)的小鼠卵巢顆粒細(xì)胞 （GC） 細(xì)胞凋亡，但其具體靶點(diǎn)尚不清楚。本研究確定了黃芩素的潛在靶點(diǎn)，以闡明黃芩素對(duì)ZEA誘導(dǎo)的卵巢損傷的機(jī)制。采用磁性納米探針法和液相色譜-串聯(lián)質(zhì)譜法，獲得小鼠卵巢GCs中黃芩素的287個(gè)靶標(biāo)。Wnt5a與黃芩素的結(jié)合自由能最低，為?8.3 kcal/mol，QRT-PCR和Western blot結(jié)果顯示，黃芩素與ZEA處理組相比，黃芩蛋白顯著增加Wnt5a和β-catenin表達(dá)，黃芩素處理組在干擾Wnt5a表達(dá)后，裂解半胱天冬酶-3表達(dá)顯著升高。Wnt5a和黃芩素的親和常數(shù)（KD）為1.7 × 10?5 M。pull-down 試驗(yàn)還表明，黃芩素可以特異性地與 Wnt5a 蛋白結(jié)合。分子對(duì)接結(jié)果顯示，黃芩素可與TRY52、GLN56和SER90在Wnt5a蛋白上形成氫鍵，Western blot試驗(yàn)證實(shí)SER90是該蛋白結(jié)合的重要位點(diǎn)。黃芩素顯著增加小鼠卵巢組織中Wnt5a和β-catenin的表達(dá)，降低cleaved-caspase-3的表達(dá)。綜上所述，黃芩素通過靶向Wnt5a對(duì)ZEA誘導(dǎo)的小鼠卵巢GCs發(fā)揮抗凋亡作用。


氨基磁珠請(qǐng)參考：氨基磁珠|高密度氨基磁珠|聚合物氨基磁性微球-生物磁珠專家 (purimagbead.com)