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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2018, Vol. 12 Issue (5) : 518-524
Oxidative stress in granulosa cells contributes to poor oocyte quality and IVF-ET outcomes in women with polycystic ovary syndrome
Qiaohong Lai1(), Wenpei Xiang2, Qing Li1, Hanwang Zhang1, Yufeng Li1, Guijin Zhu1, Chengliang Xiong2, Lei Jin1()
1. Reproductive Medicine Center, Tongji Hospital
2. Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
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The increased levels of intracellular reactive oxygen species (ROS) in granulosa cells (GCs) may affect the pregnancy results in women with polycystic ovary syndrome (PCOS). In this study, we compared thein vitro fertilization and embryo transfer (IVF-ET) results of 22 patients with PCOS and 25 patients with tubal factor infertility and detected the ROS levels in the GCs of these two groups. Results showed that the PCOS group had significantly larger follicles on the administration day for human chorionic gonadotropin than the tubal factor group (P<0.05); however, the number of retrieved oocytes was not significantly different between the two groups (P>0.05). PCOS group had slightly lower fertilization, cleavage, grade I/II embryo, clinical pregnancy, and implantation rates and higher miscarriage rate than the tubal factor group (P>0.05). We further found a significantly higher ROS level of GCs in the PCOS group than in the tubal factor group (P<0.05). The increased ROS levels in GCs caused GC apoptosis, whereas NADPH oxidase 2 (NOX2) specific inhibitors (diphenyleneiodonium and apocynin) significantly reduced the ROS production in the PCOS group. In conclusion, the increased ROS expression levels in PCOS GCs greatly induced cell apoptosis, which further affected the oocyte quality and reduced the positive IVF-ET pregnancy results of women with PCOS. NADPH oxidase pathway may be involved in the mechanism of ROS production in GCs of women with PCOS.

Keywords PCOS      ROS      granulosa cell      IVF-ET      NADPH oxidase     
Corresponding Authors: Qiaohong Lai,Lei Jin   
Just Accepted Date: 01 November 2017   Online First Date: 26 December 2017    Issue Date: 29 September 2018
 Cite this article:   
Qiaohong Lai,Wenpei Xiang,Qing Li, et al. Oxidative stress in granulosa cells contributes to poor oocyte quality and IVF-ET outcomes in women with polycystic ovary syndrome[J]. Front. Med., 2018, 12(5): 518-524.
Parameters PCOS (n = 22) Tubal (n = 25) P value
Age (year) 28.64±3.12 29.96±4.01 NS
Basal FSH (mIU/mL) 5.79±2.65 6.66±2.13 NS
Basal LH (IU/L) 5.61±3.26 5.54±3.17 NS
BMI (kg/m2) 23.21±2.85 22.12±1.89 NS
D3 antral follicle count (AFC) 21.36±8.14 13.27±6.32 <0.05
Duration of infertility (year) 3.29±1.22 3.66±2.35 NS
Tab.1  Baseline characteristics of the PCOS and tubal groups
Parameters PCOS (n = 22) Tubal (n = 25) P value
rFSH dosage (amp) 22.95±8.17 26.37±9.49 NS
rFSH duration (d) 9.86±1.94 9.51±1.88 NS
HCG day
Estradiol (pmol/L) 4873.27±3202.11 3294.87±3018.24 <0.05
Progesterone (ng/mL) 1.34±0.57 1.29±0.94 NS
≥14 mm follicles 20.23±9.18 15.21±8.73 <0.05
Endometrial thickness (mm) 12.51±2.53 11.37±2.14 NS
Oocytes retrieved 15.18±1.36 11.78±1.25 <0.05
Mature oocytes 9.09±1.23 8.57±1.01 NS
Fertilization rate (2PN) (%)
Cleavage rate (2PN) (%)
Grade I/II embryo formation (%) 39.96 48.72 NS
Clinical pregnancy rate (%) 42.98 50.67 NS
Implantation rate (%) 38.26 46.34 NS
Miscarriage rate (%) 22 15 NS
Tab.2  Ovarian stimulation characteristics and IVF results for the PCOS and tubal groups
Fig.1  Comparison of the GC ROS level and apoptosis between PCOS and tubal factor groups. (A) Isolated GCs were resuspended in DMEM medium and incubated at 25 µmol/L carboxy-H2DCF-DA in 30 min. Green fluorescence was observed under a fluorescence microscope. The GC suspension in each well of the 96-well plates was seeded, and the fluorescent intensity was measured by microplate fluorescence. (B) GC apoptosis was detected by TUNEL assay. White arrows indicate apoptotic cells. Positive cells were counted, and the difference was significant between two groups (P<0.05).
Fig.2  (A) The western blot results of NOX2 subunits in GC. The expression levels of p47phox and gp91phox protein of the two groups are not different. (B) The intracellular ROS fluorescence was detected. GCs were divided into three groups: DMSO group (10 mg/mL), DPI group (5 pmol/L), and apocynin group (20 pmol/L) and were incubated for 1 h. The cells were washed and added with 25 µmol/L carboxy-H2DCF-DA for 30 min, and the ROS level was detected. The DPI and apocynin in tubal factor group slightly inhibited the ROS levels compared with DMSO, and no differences were observed. The ROS levels in PCOS group significantly decreased after adding DPI and apocynin compared with adding DMSO (P<0.05).
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