Toxic effects of perfluorooctanoic acid on mouse oocytes
-
摘要:目的
研究全氟辛酸(Perfuorooctanoic aid，PFOA)对小鼠卵母细胞的毒性影响。
方法选用6周龄体质量相近的昆明雌性小鼠160只，分为4组，每组5个重复，每个重复8只小鼠。在适应环境7 d后开始对小鼠灌服不同剂量的PFOA，其中，对照组、低剂量组、中剂量组和高剂量组的每日剂量分别为0、 5、 10和20 mg/kg，连续灌服14 d，然后利用孕马血清促性腺激素(Pregnant mare serum gonadotropin，PMSG)和人绒毛促性腺激素(Human chorionic gonadotropin，hCG)对各组小鼠进行超数排卵，获取卵母细胞进行检测。
结果与对照组相比，PFOA中、高剂量组小鼠的卵母细胞成熟率分别下降了14.28%、28.17%；卵母细胞内活性氧含量分别升高了135%、177%；而且卵母细胞β-tubulin形态异常，染色体非整齐排列的比例分别升高了65.06%、75.60%。此外，低、中、高剂量组PFOA处理后小鼠的卵母细胞内磷酸化组蛋白H2AX(Phospho-histone H2A.X，P-H2A.X)比例分别比对照组升高了47%、133%和171%。
结论灌服PFOA可诱导小鼠卵母细胞内活性氧升高，DNA双链断裂和细胞骨架损伤，降低卵母细胞成熟率。
Abstract:ObjectiveThe purpose of this study was to investigate the toxic effect of perfuorooctanoic aid(PFOA) on mouse oocytes.
MethodOne hundred and sixty 6-week-old Kunming female mice with similar body weight were divided randomly into four groups, with five replicates in each group and eight mice in each replicate. After 7 days of adaptation, the mice were fed with different doses of PFOA for 14 days. The daily doses for control, low, medium, and high dose groups were 0, 5, 10 and 20 mg/kg respectively. Then the mice were superovulated with injection of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG), and the oocytes were obtained for detection.
ResultCompared to the control group, the oocyte maturation rates of mice in the middle and high dose groups decreased by 14.28% and 28.17% respectively, while the contents of reactive oxygen species in oocytes increased by 135% and 177% respectively. The proportions of β-tubulin with abnormal morphology and irregular chromosomes in oocytes increased by 65.06% and 75.60% respectively. In addition, the proportions of (Phospho-histone H2A.X, P-H2A.X) in oocytes of low, medium and high dose groups increased by 47%, 133% and 1.71% respectively compared with the control.
ConclusionMice fed with PFOA may decrease their oocyte maturation rate by inducing the increase of reactive oxygen species, DNA double strand break and cytoskeleton damage in oocytes.
-
Keywords:
- Perfuorooctanoic aid /
- Oocyte /
- Reactive oxygen species /
- DNA damage /
- Cytoskeleton
-
-
![]()
图 1 不同剂量 PFOA暴露的卵母细胞内活性氧(ROS)荧光图像
Figure 1. Fluorescence images of reactive oxygen species (ROS) in oocytes exposed to different doses of PFOA
a: 0 (CK)；b: 5 mg/kg；c: 10 mg/kg；d: 20 mg/kg
![]()
图 2 不同剂量 PFOA暴露的卵母细胞内活性氧(ROS)荧光强度
柱子上方的不同小写字母表示差异显著(P<0.05, LSD法)
Figure 2. The fluorescence intensities of reactive oxygen species (ROS) in oocytes exposed to different doses of PFOA
Different lowercase letters on bars indicate significant differences(P< 0.05, LSD method)
![]()
图 3 不同剂量PFOA暴露的卵母细胞内P-H2A.X荧光图像
Figure 3. Fluorescence images of P-H2A.X in oocytes exposed to different doses of PFOA
![]()
图 4 不同剂量PFOA暴露的卵母细胞DNA损伤比例
柱子上方的不同小写字母表示差异显著（P<0.05, LSD法）
Figure 4. DNA damage rates of oocytes exposed to different doses of PFOA
Different lowercase letters on bars indicate significant differences (P< 0.05, LSD method)
![]()
图 5 不同剂量PFOA暴露的卵母细胞骨架荧光图像
Figure 5. Fluorescence images of cytoskeleton of oocytes exposed to different doses of PFOA
![]()
图 6 不同剂量PFOA暴露的卵母细胞骨架异常比例
柱子上方的不同小写字母表示差异显著（P<0.05, LSD法）
Figure 6. The proportions of abnormal cytoskeleton in oocytes exposed to different doses of PFOA
Different lowercase letters on bars indicate significant differences (P< 0.05, LSD method)
表 1 PFOA对小鼠卵母细胞成熟率的影响
Table 1 Effect of PFOA on maturation of mouse oocytes
每日剂量/(mg·kg−1)
Daily dose细胞总数
Total cell count第一极体排出率1)/%
Discharge rate of first polar body0(CK) 108 91.23±0.010a 5 111 85.50±0.123a 10 104 78.20±0.015b 20 101 65.53±0.372c 1) 同列数据后的不同小写字母表示差异显著(P<0.05, LSD法)
1)Different lowercase letters in the same column indicate significant differences(P<0.05, LSD method)
下载: 导出CSV
-
[1] GIESY J P, KANNAN K, JONES P D. Global biomonitoring of perfluorinated organics[J]. Scientific WorldJournal, 2001, 1: 627-629.
[2] KOSKELA A, FINNILA M A, KORKALAINEN M, et al. Effects of developmental exposure to perfluorooctanoic acid (PFOA) on long bone morphology and bone cell differentiation[J]. Toxicology and Applied Pharmacology, 2016, 301: 14-21. doi: 10.1016/j.taap.2016.04.002
[3] FROMME H, TITTLEMIER S A, VOLKEL W, et al. Perfluorinated compounds: Exposure assessment for the general population in Western countries[J]. International Journal of Hygiene & Environmental Health, 2009, 212(3): 239-270.
[4] 夏慧, 敖俊杰, 袁涛. 室内灰尘中全氟化合物的污染状况与人体暴露水平评估[J]. 生态毒理学报, 2016, 11(2): 223-230. [5] 杜国勇, 蒋小萍, 卓丽, 等. 长江流域重庆段水体中全氟化合物的污染特征及风险评价[J]. 生态环境学报, 2019, 28(11): 2266-2272. [6] 陈诗艳, 仇雁翎, 朱志良, 等. 土壤中全氟和多氟烷基化合物的污染现状及环境行为[J]. 环境科学研究, 2021, 34(2): 468-478. [7] JOGSTEN I E, PERELLO G, LLEBARIA X, et al. Exposure to perfluorinated compounds in Catalonia, Spain, through consumption of various raw and cooked foodstuffs, including packaged food[J]. Food and chemical toxicology, 2009, 47(7): 1577-1583. doi: 10.1016/j.fct.2009.04.004
[8] BACH C C, VESTED A, JORGENSEN K T, et al. Perfluoroalkyl and polyfluoroalkyl substances and measures of human fertility: A systematic review[J]. CRC Critical Reviews in Toxicology, 2016, 46(9): 735-755. doi: 10.1080/10408444.2016.1182117
[9] ZHANG S, TAN R, PAN R, et al. Association of perfluoroalkyl and polyfluoroalkyl substances with premature ovarian insufficiency in Chinese women[J]. The Journal of Clinical Endocrinology and Metabolism, 2018, 103(7): 2543-2551. doi: 10.1210/jc.2017-02783
[10] CHEN Y, ZHOU L, XU J, et al. Maternal exposure to perfluorooctanoic acid inhibits luteal function via oxidative stress and apoptosis in pregnant mice[J]. Reproductive Toxicology, 2017, 69: 159-166. doi: 10.1016/j.reprotox.2017.02.010
[11] DU G, HU J, HUANG Z, et al. Neonatal and juvenile exposure to perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS): Advance puberty onset and kisspeptin system disturbance in female rats[J]. Ecotoxicology and Environmental Safety, 2019, 167: 412-421. doi: 10.1016/j.ecoenv.2018.10.025
[12] KANG Q, GAO F, ZHANG X, et al. Nontargeted identification of per- and polyfluoroalkyl substances in human follicular fluid and their blood-follicle transfer[J]. Environment International, 2020, 139: 105686. doi: 10.1016/j.envint.2020.105686
[13] RASHID F, RAMAKRISHNAN A, FIELDS C, et al. Acute PFOA exposure promotes epigenomic alterations in mouse kidney tissues[J]. Toxicology Reports, 2020, 7: 125-132. doi: 10.1016/j.toxrep.2019.12.010
[14] XU W, LI L, SUN J, et al. Putrescine delays postovulatory aging of mouse oocytes by upregulating PDK4 expression and improving mitochondrial activity[J]. Aging (Albany NY), 2018, 10(12): 4093-4106.
[15] LOPEZ-ARELLANO P, LOPEZ-ARELLANO K, LUNA J, et al. Perfluorooctanoic acid disrupts gap junction intercellular communication and induces reactive oxygen species formation and apoptosis in mouse ovaries[J]. Environmental Toxicology, 2019, 34(1): 92-98. doi: 10.1002/tox.22661
[16] YANG L L, ZHAO Y, LUO S M, et al. Toxic effects and possible mechanisms of hydrogen sulfide and/or ammonia on porcine oocyte maturation in vitro[J]. Toxicology Letters, 2018, 285: 20-26. doi: 10.1016/j.toxlet.2017.12.019
[17] YU Y, CUI Y, NIEDERNHOFER L J, et al. Occurrence, biological consequences, and human health relevance of oxidative stress-induced DNA damage[J]. Chemical Research in Toxicology, 2016, 29(12): 2008-2039. doi: 10.1021/acs.chemrestox.6b00265
[18] DING Z M, ZHANG S X, JIAO X F, et al. Doxorubicin exposure affects oocyte meiotic maturation through DNA damage induced meiotic arrest[J]. Toxicological Sciences, 2019, 171(2): 359-368.
[19] DING Z M, JIAO X F, WU D, et al. Bisphenol AF negatively affects oocyte maturation of mouse in vitro through increasing oxidative stress and DNA damage[J]. Chemico-Biological Interactions, 2017, 278: 222-229. doi: 10.1016/j.cbi.2017.10.030
[20] JEONG P S, LEE S, PARK S H, et al. Butylparaben is toxic to porcine oocyte maturation and subsequent embryonic development following in vitro fertilization[J]. International Journal of Molecular Sciences, 2020, 21(10): 3692. doi: 10.3390/ijms21103692
[21] JIAO X, DING Z, MENG F, et al. The toxic effects of Fluorene-9-bisphenol on porcine oocyte in vitro maturation[J]. Environmental Toxicology, 2020, 35(2): 152-158. doi: 10.1002/tox.22851
[22] TRANFO G, CAPOROSSI L, PACI E, et al. Urinary phthalate monoesters concentration in couples with infertility problems[J]. Toxicology Letters, 2012, 213(1): 15-20. doi: 10.1016/j.toxlet.2011.11.033
[23] BENNABI I, TERRET M E, VERLHAC M H. Meiotic spindle assembly and chromosome segregation in oocytes[J]. Journal of Cell Biology, 2016, 215(5): 611-619. doi: 10.1083/jcb.201607062
[24] AZOURY J, VERLHAC M H, DUMONT J. Actin filaments: Key players in the control of asymmetric divisions in mouse oocytes[J]. Biology of the Cell, 2009, 101(2): 69-76. doi: 10.1042/BC20080003
