青春期前双酚A暴露对雌性大鼠卵巢组织中mRNA分子表达谱的影响

doi:10.13481/j.1671-587X.20210609

摘要/Abstract

摘要： 目的

研究双酚A（BPA）对青春期前雌性大鼠卵巢组织中mRNA分子表达谱的影响，探讨BPA对卵巢结构和功能的潜在影响。

方法

32只雌性SD大鼠随机分为对照组和低、中及高剂量BPA组，每组8只。低、中和高剂量BPA组大鼠分别给予50、100和200 mg·kg^－1BPA，对照组大鼠给予等体积乙醇-玉米油（1∶100）。10 d后HE染色观察各组大鼠卵巢组织形态表现，RNA测序（RNA-seq）和生物信息学分析预测高剂量BPA组和对照组大鼠的差异表达基因及其生物学功能，实时荧光定量PCR（RT-qPCR）法检测各组大鼠卵巢组织中Hapln3、Nr4a1、Arc、Ccl2、Ccn5、Egr2和Rab33a mRNA表达水平。

结果

HE染色，与对照组比较，低和中剂量BPA组大鼠卵巢组织未见明显异常，高剂量BPA组大鼠卵巢组织中原始卵泡、窦前卵泡、窦状卵泡数减少，闭锁卵泡数增加，出现囊状卵泡等改变。RNA-seq检测，对照组和高剂量BPA组大鼠卵巢组织中有14个差异表达基因。生物信息学分析预测该14个mRNA分子参与了生殖、代谢和细胞凋亡等多种生物学过程及p53、MAPK和PI3K-Akt等信号通路。 RT-qPCR法，与对照组比较，高剂量BPA组大鼠卵巢组织中Hapln3、Nr4a1、Arc、Ccl2、Ccn5和Egr2 mRNA表达水平降低（P<0.05），Rab33a mRNA表达水平升高（P<0.05）。

结论

BPA可引起青春期前雌性大鼠卵巢结构和卵巢组织中mRNA分子表达谱的变化，这可能会影响卵巢组织的结构和功能。

关键词: 双酚A, 环境内分泌干扰物, 卵巢组织, mRNA, 大鼠，SD, 青春期前

Abstract: Objective

To study the effect of bisphenol A （BPA） on the mRNA molecule expression profile in ovarian tissue of the prepubertal female rats， and to explore the potential effect of BPA on the structure and function of ovarian tissue.

Methods

A total of 32 female SD rats were randomly divided into control group and low，middle，and high doses of BPA groups，and there were 8 rats in each group.The rats in low，middle，and high doses of BPA groups were given 50，100，and 200 mg·kg^－1 BPA，and the rats in control group were given ethanol-corn oil（1∶100）. HE staining was used to observe the morphology of ovarian tissue of the rats in various groups. RNA sequencing （RNA-seq） and bioinformatic analyses were used to predict the differential expression genes and their biological functions between high dose of BPA group and control group. The mRNA expression levels of Hapln3， Nr4a1， Arc， Ccl2， Ccn5， Egr2，and Rab33a in ovarian tissue of the rats in various groups were detected by Real-time fluorescence quantitative PCR （RT-qPCR）method.

Results

The HE staining results showed that compared with control group， there were no obvious abnormalities in ovarian tissue of the rats in low and middle doses of BPA groups. The number of primordial follicles， pre sinus follicles and sinus follicles in ovarian tissue of the rats in high dose of BPA group were decreased， the number of atretic follicles was increased， and the cystic follicles were appeared.The RNA-seq detection results showed that there were 14 differential expression genes in ovarian tissue of the rats in control group and high dose of BPA group. The bioinformatics analysis results showed that 14 mRNA molecules were involved in a variety of biological processes such as reproduction， metabolism and apoptosis， as well as p53， MAPK， PI3K-Akt，and other signal pathways. Compared with control group， the expression levels of Hapln3， Nr4a1， Arc， Ccl2， Ccn5，and Egr2 mRNA in ovarian tissue of the rats in high dose of BPA group were decreased （P<0.05），and the expression level of Rab33a mRNA was increased （P<0.05）.

Conclusion

BPA changes the mRNA molecule expression profile in ovarian tissue of the prepubertal female rats， which may potentially influence the structure and function of ovarian tissue.

Key words: bisphenol a, endocrine disrupting chemicals, ovarian tissue, mRNA, rat，SD, prepubertal

中图分类号:

R994.6

张雪莹,郑连文,邹颖刚,周振环. 青春期前双酚A暴露对雌性大鼠卵巢组织中mRNA分子表达谱的影响[J]. 吉林大学学报(医学版), 2021, 47(6): 1407-1414.

Xueying ZHANG,Lianwen ZHENG,Yinggang ZOU,Zhenhuan ZHOU. Effect of bisphenol A exposure on mRNA molecule expression profile in ovarian tissue of prepubertal female rats[J]. Journal of Jilin University(Medicine Edition), 2021, 47(6): 1407-1414.

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参考文献 29

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Group	Body weight
Group	PND28	PND32	PND36	PND41
Control	100.43±0.75	121.80±1.11	141.34±1.69	156.11±3.20
BPA
Low dose	103.43±2.01	126.88±2.91	146.98±3.22	157.03±4.04
Middle dose	101.79±1.36	122.61±1.79	143.44±2.27	157.85±3.08
High dose	98.95±1.20	121.79±1.37	144.01±1.77	157.16±2.84

Group	Ovarian weight （m/mg）	Ovarian coefficient（η/%）
Control	20.480±0.759	0.130±0.004
BPA
Low dose	19.350±0.974	0.120±0.006
Middle dose	18.310±1.333	0.120±0.008
High dose	14.180±1.282 ^*	0.090±0.007 ^*

Group	Hapln3	Nr4a1	Arc	Ccl2	Ccn5	Egr2	Rab33a
Control	1.014 7±0.117 3	1.014 7±0.117 3	1.014 7±0.117 3	1.014 7±0.117 3	1.014 7±0.117 3	1.014 7±0.117 3	1.014 7±0.117 3
High dose of BPA	0.078 5±0.014 4^*	0.078 5±0.014 4^*	0.078 5±0.014 4^*	0.078 5±0.014 4^*	0.078 5±0.014 4^*	0.078 5±0.014 4^*	0.078 5±0.014 4^*

KEGG Pathway Classification	Related Genes
Cytokine-cytokine receptor interaction	Inhba, Ccl2
Apoptosis - multiple species	Pmaip1
Aldosterone synthesis and secretion	Nr4a1
NOD-like receptor signaling pathway	Ccl2
p53 signaling pathway	Pmaip1
TGF-beta signaling pathway	Inhba
TNF signaling pathway	Ccl2
Signaling pathways regulating pluripotency of stem cells	Inhba
Apoptosis	Pmaip1
Chemokine signaling pathway	Ccl2
MAPK signaling pathway	Nr4a1
PI3K-Akt signaling pathway	Nr4a1

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