毛蕊异黄酮缓解双酚A损伤小鼠睾丸功能的机理研究

doi:10.16431/j.cnki.1671-7236.2025.09.016

摘要/Abstract

摘要： 【目的】毛蕊异黄酮(calycosin，CAL)是从黄芪等植物中提取的天然异黄酮化合物，具有抗氧化、抗炎等多重药理活性。试验旨在探究CAL缓解环境内分泌干扰物双酚A(bisphenol A，BPA)对睾丸功能损伤的作用机制。【方法】选取30只6周龄雄性昆明小鼠作为模型动物，分为空白对照组、BPA组和BPA+CAL组，每组10个重复。BPA、BPA+CAL组小鼠每天灌胃50 mg/kg BPA，1周后BPA+CAL组小鼠每天灌服40 mg/kg CAL，空白组小鼠每天灌胃同等体积玉米油。期间每天测量小鼠体重，灌胃处理后第22天，各组小鼠眼球采血，通过ELISA检测血清中睾酮(testosterone，T)、雌二醇(estradiol，E₂)含量，脱颈处死后分离睾丸组织进行HE染色，提取睾丸组织蛋白样品，对内质网应激相关蛋白(Bip、ATF6、p-PERK、eIF2α和ATF4)表达量进行Western blotting分析。【结果】 ①BPA组小鼠体重在部分试验天数显著低于空白对照组(P＜0.05)，而BPA+CAL组小鼠体重有高于BPA组的趋势(P＞0.05)。②BPA组小鼠睾丸重量显著低于空白对照组(P＜0.05)，CAL显著改善了BPA导致的睾丸重量降低(P＜0.05)。与空白对照组相比，BPA组小鼠睾丸组织中存在明显的损伤性结构变化，但CAL治疗后小鼠睾丸组织结构损伤较BPA组明显改善。③与空白对照组相比，BPA组小鼠血清中T激素水平显著升高(P＜0.05)，而E₂激素水平显著降低(P＜0.05)；与BPA组相比，BPA+CAL组小鼠血清中T激素水平显著降低(P＜0.05)，而E₂激素水平显著升高(P＜0.05)。④Western blotting分析结果显示，BPA组小鼠睾丸组织中Bip、ATF6、p-PERK、eIF2α和ATF4等内质网应激相关蛋白表达水平显著高于空白对照组(P＜0.05)，而BPA+CAL组小鼠上述蛋白表达水平与BPA组相比有下调趋势，表明CAL可能通过抑制内质网应激反应减轻BPA对睾丸组织的生殖损害。【结论】 CAL可缓解BPA暴露导致的小鼠睾丸组织损伤，调节性激素稳态平衡，抑制睾丸组织内质网应激反应，进而改善小鼠生殖功能。

关键词: 毛蕊异黄酮; 双酚A; 睾丸损伤; 内质网应激

Abstract: 【Objective】 Calycosin (CAL),a naturally occurring isoflavone compound derived from medicinal plants such as Astragalus membranaceus,exhibits diverse pharmacological properties,including antioxidant and anti-inflammatory activities.This study was aimed to investigate the mechanism of CAL alleviated testicular dysfunction induced by bisphenol A (BPA),a pervasive environmental endocrine-disrupting chemical. 【Method】 Thirty 6-week-old male Kunming mice were randomly assigned into three groups (n=10 per group):Control group (vehicle-treated with corn oil),BPA group (oral gavage of 50 mg/kg BPA daily),and BPA+CAL group (received 50 mg/kg BPA daily for 1 week followed by 40 mg/kg CAL daily).Body weight was monitored throughout the experiment.On day 22 post-intervention,blood samples were collected via retro-orbital puncture for ELISA quantification of the levels of testosterone (T) and estradiol (E₂) in serum.Mice were euthanized by cervical dislocation,and testicular tissues were collected for histopathological evaluation via HE staining.Protein extracts from testicular tissues were subjected to Western blotting for analyzing endoplasmic reticulum stress markers,including Bip,ATF6,p-PERK,eIF2α,and ATF4. 【Result】 ①Compared with control group,mice in BPA group exhibited a significant reduction of body weight in some experimental days (P＜0.05),while BPA+CAL-treated group showed a non-significant upward trend in body weight relative to BPA group (P＞0.05).②Testicular weight in BPA group was significantly lower than that of control group (P＜0.05),but this decline was significantly ameliorated by CAL intervention (P＜0.05).Compared with control group,there was significant structural damage in testicular tissue of mice in BPA group,but the structural damage was significantly improved after CAL treatment relative to BPA group.③ Compared with control group,the T level of serum of mice in BPA group was significantly increased (P＜0.05),while the E₂ level was significantly decreased (P＜0.05).Compared with BPA group,the T level of serum of mice in BPA+CAL group was significantly decreased (P＜0.05),while the E₂ level was significantly increased (P＜0.05).④Western blotting analysis results demonstrated that the expression of stress-related proteins (Bip,ATF6,p-PERK,eIF2α,and ATF4) in testicular tissue of mice in BPA group were significantly higher than those of control group (P＜0.05).However,CAL treatment attenuated this response,showing a downregulation trend of these proteins compared with BPA group,suggesting that CAL mitigated BPA-induced testicular injury by suppressing endoplasmic reticulum stress pathways. 【Conclusion】 CAL could mitigate BPA-induced testicular injury in mice,regulate sex hormone homeostasis,inhibit endoplasmic reticulum stress response in testicular tissue,and thereby improve reproductive function in mice.

Key words: calycosin; bisphenol A; testicular injury; endoplasmic reticulum stress

中图分类号:

S852.2

魏雨轩, 张家希, 崔楠, 曹琪, 吴琼. 毛蕊异黄酮缓解双酚A损伤小鼠睾丸功能的机理研究[J]. 中国畜牧兽医, 2025, 52(9): 4173-4181.

WEI Yuxuan, ZHANG Jiaxi, CUI Nan, CAO Qi, WU Qiong. Mechanistic Study on Calycosin in Ameliorating Bisphenol A-induced Testicular Dysfunction in Mice[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4173-4181.

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