一氧化氮在小鼠囊胚发育和孵化中的调控作用

doi:10.3881/j.issn.1000-503X.2015.05.003

Acta Academiae Medicinae Sinica ›› 2015, Vol. 37 ›› Issue (5): 501-507.doi: 10.3881/j.issn.1000-503X.2015.05.003

• Orginal Article • Previous Articles Next Articles

Regulatory Role of Nitric Oxide in Development and Hatching of Mouse Blastocysts

Xiao-yan PAN¹, Zhi-xin LI¹(), Xi-yan WANG¹, Xue-nan WANG², Jian-xin SUN¹, Meng-tong ZANG¹, Wen-jun LI¹, Hong-he WANG¹, Zhao-hua DOU¹

¹Department of Histology and Embryology,Jilin Medical College,Jilin,Jilin 132013,China
²Reproductive Medicine Center,the Affiliated Hospital of Jining Medical College,Jining,Shandong 272029,China

Received:2014-11-17 Online:2015-10-15 Published:2015-10-31
Supported by:
Supported by the “Twelfth Five-Year” Scientific and Technological Research Project of Education Department of Jilin Province [吉教科合字(2013)第349号],the Natural Science Foundation of Shandong Province(ZR2011HL005),the Scientific and Technological Research Project of Jilin Province [201201077],the College Students’ Scientific Research Fund of Jilin Medical College [吉医学科字(2012)第04号],and the College Students’ Innovative and Entrepreneurial Training Program of Jilin Province[(2013)第850号]

Abstract

Abstract:

Objective To determine the regulatory role and mechanism of nitric oxide (NO) in the development and hatching of mouse blastocysts. Methods The Kunming female mice were superovulated and then mated with mature male mice. On the day 2.5 of their pregnancy,morulae were flushed from their uterine horns with culture media. Morulae were cultured in different concentrations of N-nitro-L arginine methyl ester (L-NAME),sodium nitroprusside (SNP),or the combination of L-NAME and SNP in culture media for 48 hours. The development and hatching of blastocysts were examined on day 4 and day 5 and the total numbers of blastocyst cells and cysteinyl aspartate specific proteinase 3 (caspase 3) were observed under confocal laser scanning microscope. Results With the increase of the concentration of L-NAME or SNP,the hatching rate of blastocysts and the total number of blastocyst cells were significantly reduced. The addition of 10 nmol/L SNP in culture media with 5 mmol/L L-NAME significantly increased the development of blastocysts and promoted hatching of blastocysts. However,with increase of SNP concentration in culture media with 5 mmol/L L-NAME,the development and hatching rates of blastocysts were significantly decreased. L-NAME had no obvious effect on the expression of active caspase 3 in blastocyst cells. However,when being above 500 nmol/L,SNP significantly increased the expression of caspase 3 in blastocyst cells. Conclusions NO plays an important role in development and hatching of mouse blastocysts. Excessively high or low NO can damage the division of blastomeres,resulting in the failure of the blastocyst development and hatching. Also,excessively high NO can lead to the apoptosis of the blastocyst cells.

Key words: mouse, nitric oxide, blastocyst development, blastocyst hathing, apoptosis

CLC Number:

R321.2

Xiao-yan PAN, Zhi-xin LI, Xi-yan WANG, Xue-nan WANG, Jian-xin SUN, Meng-tong ZANG, Wen-jun LI, Hong-he WANG, Zhao-hua DOU. Regulatory Role of Nitric Oxide in Development and Hatching of Mouse Blastocysts[J].Acta Academiae Medicinae Sinica, 2015, 37(5): 501-507.

Figures/Tables 8

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References 18

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分组 Group	桑葚胚的数量 Number of morulae	D4囊胚发育率 Percentage of blastocysts on Day 4	D5囊胚孵化率 Percentage of hatched blastocyst on Day 5
对照组Control group	172	92.19±3.80	92.26±7.69
125 μmol/L L-NAME	156	85.88±4.30	86.52±5.50
250 μmol/L L-NAME	130	69.67±3.21^a	72.81±4.44^a
500 μmol/L L-NAME	147	34.71±6.35^ab	38.57±5.34^ab
5 mmol/L L-NAME	73	11.15±4.33^abc	0.00±0.00^abc

分组 Group	桑葚胚的数量 Number of morulae	D4囊胚发育率 Percentage of blastocysts on D4	D5囊胚孵化率 Percentage of hatched blastocyst on D5
对照组Control group	152	92.19±3.80	91.51±4.65
10 nmol/L SNP	121	88.68±3.78	87.10±6.74
100 nmol/L SNP	119	53.19±6.36^a	57.60±6.23^a
500 nmol/L SNP	106	28.72±3.93^ab	25.77±5.66^ab
2 μmol/L SNP	124	7.89±2.60^abc	0.00±0.00^abc

分组 Group	桑葚胚数量 Number of morulae	D4囊胚发育率 Percentage of blastocysts on D4	D5囊胚孵化率 Percentage of hatched blastocyst on D5
5 mmol/L L-NAME	73	11.15±4.33	0.00±0.00
5 mmol/L L-NAME+10 nmol/L SNP	123	52.46±9.40^a	47.00±4.32^a
5 mmol/L L-NAME+500 nmol/L SNP	114	31.36±5.58^ab	36.64±4.73^ab
5 mmol/L L-NAME+2 μmol/L SNP	102	17.10±2.39	16.10±4.84^abc

分组 Group	囊胚的数量 Number of blastocysts	单个囊胚的细胞总数 Total cell number of blastocysts	Caspase 3的平均荧光强度 Mean fluorescence intensity of caspase 3
对照组Control group	13	65.33±5.51	0.54±0.09
125 μmol/L L-NAME	10	62.33±3.51	0.57±0.08
500 μmol/L L-NAME	11	43.33±3.06^a	0.62±0.11
5 mmol/L L-NAME	10	32.00±3.61^ab	0.56±0.07

分组 Group	囊胚的数量 Number of blastocysts	单个囊胚的细胞总数 Total cell number of blastocyst	Caspase 3的平均荧光强度 Mean fluorescence intensity of caspase 3
对照组Control group	13	65.33±5.51	0.54±0.09
10 nmol/L SNP	12	62.33±2.08	0.56±0.07
500 nmol/L SNP	12	44.33±3.06^a	6.53±0.79^a
2 μmol/L SNP	10	33.00±3.00^ab	11.15±0.90^ab

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