番茄红素缓解双酚A致小鼠生殖毒性的研究

doi:10.16431/j.cnki.1671-7236.2024.11.037

摘要/Abstract

摘要： 【目的】研究番茄红素对双酚A(bisphenol A,BPA)致小鼠生殖毒性的缓解作用。【方法】将30只昆明鼠孕鼠随机分为3组，每组10只。对照组孕鼠妊娠第1～7天灌胃生理盐水同时正常饲喂；BPA处理组孕鼠妊娠第1～7天灌胃生理盐水，妊娠第8～14天混饲500 mg/kg BPA；番茄红素处理组孕鼠妊娠第1～7天灌胃20 mg/kg番茄红素，妊娠第8～14天混饲500 mg/kg BPA，待孕鼠自然分娩，观察记录F1代小鼠存活数。F1代小鼠正常饲喂至8周剖杀，采集睾丸、卵巢组织，通过HE染色观察睾丸、卵巢组织结构变化，利用ELISA法检测F1代小鼠血清睾酮(T)、促卵泡素(FSH)、雌二醇(E₂)及黄体生成素(LH)水平；使用免疫组化法和免疫荧光法检测F1代小鼠睾丸及卵巢中B淋巴细胞瘤-2(Bcl-2)和B淋巴细胞瘤-2相关X蛋白(Bax)的表达。【结果】 BPA处理后，F1代小鼠存活率为62.50%，明显低于对照组(83.65%)和番茄红素处理组(81.25%)。与对照组相比，BPA处理组F1代小鼠血清中生殖激素T、FSH、LH水平均显著降低(P＜0.05)，E₂水平显著升高(P＜0.05)；小鼠睾丸组织曲细精管中细胞排列疏松、紊乱，间质细胞数量减少、管腔变大、管壁变薄；卵巢组织中颗粒细胞过度凋亡，细胞排列疏松。与BPA处理组相比，番茄红素处理组F1代小鼠血清中T、FSH、LH水平显著升高(P＜0.05)，E₂水平显著降低(P＜0.05)；小鼠睾丸组织结构完整，生精小管内各级生精细胞排列紧密、整齐有序；卵巢组织中可见不同发育阶段的卵泡，颗粒细胞排列整齐，少量颗粒细胞发生调亡。免疫组化和免疫荧光检测结果显示，Bax、Bcl-2蛋白表达于睾丸间质细胞及卵巢颗粒细胞中。与对照组相比，BPA处理组F1代小鼠卵巢组织中Bax蛋白表达量显著升高(P＜0.05)，睾丸和卵巢组织中Bcl-2蛋白表达量均显著降低(P＜0.05)。与BPA处理组相比，番茄红素处理组F1代小鼠卵巢组织中Bax蛋白表达量显著降低(P＜0.05)，睾丸和卵巢组织中Bcl-2蛋白表达量均显著升高(P＜0.05)。【结论】番茄红素通过缓解BPA引起的生殖激素水平紊乱，改善睾丸及卵巢结构，平衡凋亡与抗凋亡水平，从而颉颃BPA对小鼠的生殖损伤，研究结果为番茄红素的临床应用提供了试验依据。

关键词: 番茄红素; 双酚A(BPA); 小鼠; 生殖毒性

Abstract: 【Objective】 The aim of this study was to investigate the alleviation effect of lycopene on reproductive toxicity induced by bisphenol A (BPA) in mice. 【Method】 30 pregnant Kunming mice were randomly divided into 3 groups with 10 mice in each group.Mice in control group were gavaged with normal saline from days 1 to 7 of gestation and received conventional feeds.Mice in BPA treatment group were gavaged with normal saline on days 1 to 7 of gestation,and 500 mg/kg BPA on days 8 to 14 of gestation.Mice in lycopene treatment group were gavaged with 20 mg/kg lycopene on days 1 to 7 of gestation,and 500 mg/kg BPA on days 8 to 14 of gestation，with natural birth allowed.The survival number of F1 generation mice was observed and recorded.F1 generation mice were normally fed to 8 weeks and dissected.Testis and ovary were collected,and the structural changes of testicular and ovarian tissues were observed by HE staining.Testosterone (T),follicle-stimulating hormone (FSH),estradiol (E₂) and luteinizing hormone (LH) levels of serum in F1 generation mice were detected by ELISA.The expression of B lymphoblastoma-2 (Bcl-2) and B lymphoblastoma-2-associated X protein (Bax) in testis and ovaries of F1 generation mice were detected by immunohistochemistry and immunofluorescence techniques. 【Result】 After BPA treatment,the survival rate of F1 generation mice was 62.50%,which was significantly lower than that of control group (83.65%) and lycopene treatment group (81.25%).Compared with control group,the levels of reproductive hormones T,FSH and LH in F1 generation mice in BPA treatment group were significantly decreased (P＜0.05),and E₂ level was significantly increased (P＜ 0.05).The arrangement of cells in seminiferous tubule of mouse testis was loose and disordered,the number of interstitial cells was reduced,the lumen was enlarged and the tube wall was thin.The granulosa cells in ovarian were overapoptosis and the cell arrangement was loose.Compared with BPA treatment group,T,FSH and LH levels in F1 generation mice in lycopene treatment group were significantly increased (P＜0.05),while E₂ level was significantly decreased (P＜ 0.05).The structure of mouse testis was complete,and spermatogenic cells at all levels of spermatogenic tubule were arranged closely and neatly.Follicles at different developmental stages were observed in ovary，the granulosa cells were arranged neatly,and a few granulosa cells were regulated and died.The results of immunohistochemistry and immunofluorescence showed that Bax and Bcl-2 proteins were expressed in testicular stromal cells and ovarian granulosa cells.Compared with control group,the expression of Bax protein in ovary of F1 generation mice in BPA treatment group was significantly increased (P＜ 0.05),and the expression of Bcl-2 protein in testes and ovaries was significantly decreased (P＜0.05).Compared with BPA treatment group,the expression of Bax protein in ovaries of F1 generation mice in lycopene treatment group was significantly decreased (P＜0.05)，and the expression of Bcl-2 protein in testes and ovaries was significantly increased (P＜0.05). 【Conclusion】 Lycopene could alleviate the disturbance of reproductive hormone level caused by BPA,improve the structure of testis and ovary,balance apoptosis and anti-apoptosis levels,and thus antagonize the reproductive damage of mice caused by BPA.The results provided experimental basis for the clinical application of lycopene.

Key words: lycopene; bisphenol A (BPA); mice; reproductive toxicity

中图分类号:

S859.7

陈玉明, 张晶晶, 赵媛慧, 付国爽, 王彤, 马双. 番茄红素缓解双酚A致小鼠生殖毒性的研究[J]. 中国畜牧兽医, 2024, 51(11): 5023-5032.

CHEN Yuming, ZHANG Jingjing, ZHAO Yuanhui, FU Guoshuang, WANG Tong, MA Shuang. Alleviation of Lycopene on Reproductive Toxicity Induced by Bisphenol A in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(11): 5023-5032.

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