| Objective: To investigate the expression and significance of autophagy-related genes Beclin1 m RNA and Atg5 m RNA in Corona radiata cells and Cumulus oophorus cells of patients with polycystic ovary syndrome(PCOS)and non-polycystic ovarian syndrome,providing a new clue for the study of molecular mechanism underlying the development and progression of polycystic ovary syndrome.Methods: The standard of control group was as follows: The woman’s age was less than 35 years old and menstrual regularity;The main reason for the poor quality of semen need intracytoplasmic sperm injection assisted pregnancy;No pelvic surgery;No endometriosis;Physical examination is normal;No alcohol,tobacco hobbies and drug history.Polycystic ovary syndrome group(According to the 2003 Rotterdam standard): The woman’s age was less than 35 years old;The main reason for the poor quality of semen need intracytoplasmic sperm injection assisted pregnancy;No pelvic surgery;No endometriosis;Physical examination is normal;No alcohol,tobacco hobbies and drug history;sparse ovulation or anovulation;Ultrasound showed there are more than 12 follicles with diameter of 9mm in bilateral ovaries;With or without the clinical manifestations of hyperandrogenism and / or hyperandrogenism(With the exception of other diseases that may cause hyperandrogenism).According to the above criteria patients were received for the first time to receive assisted pregnancy in the Second Hospital of Hebei Medical University,Department of Reproductive Medicine during the March 1,2016 to June 30,2016 and transplanted embryos with fresh cycles.Collecting 17 patients in polycystic ovary syndrome group and 25 patients in the control group of abandoned granulocyte cells and the corresponding clinical data:Age,The body mass index(BMI),Promotion plan,The levels of serum luteinizing hormone(LH),estrogen(E2),progesterone(P)on the trigger day,Obtaining the number of oocytes,Fertilization rate,Whether pregnancy(Pregnancy criteria is that the ultrasound prompted intrauterine pregnancy and detected germ and cardiac beat in the first 28 days after transplantation).The expression of Beclin1 m RNA and Atg5 m RNA in granulosa cells was measured by RT-PCR(Realtime Polymerase Chain Reaction).According to the original test results of fluorescence quantitative analysis,the relative quantitative results of the target genes were calculated according to the relative quantitative formula of 2-ΔΔct.SPSS 17.0 statistical analysis software was used for data processing.The metrological data according to the normal distribution are expressed as mean ± standard deviation.Metrics that do not fit the normal distribution are expressed as median(Interquartile range).The t-test was used to compare two samples with normal distribution and variance homogeneity.The two samples that do not fit the normal distribution are compared by the rank sum test.The count data were compared with the X2 test.Pearson correlation is used to correlate the two data with normal distribution,and Spearman rank correlation analysis is used for those who did not meet the condition.P value less than 0.05 was considered as statistically significant.Results:1 Beclin1 m RNA and Atg5 m RNA were expressed in both groups.The relative expression levels of Beclin1 m RNA and Atg5 m RNA in the PCOS group were 1.612 ± 1.068 and 0.990(0.96),respectively.The expression of Beclin1 m RNA in PCOS group was significantly higher than that in control group(P <0.05).2 The clinical data of the two groups were analyzed.Compared with the control group,the BMI of the PCOS group was higher,and that was statistically significant(P <0.05).Serum LH in the control group and PCOS group were 1.910(1.140),1.190(0.620),respectively.And it was higher in the control group(P <0.05).The pregnancy rate of the two groups(Pregnancy number / total number of cases in this group):the control group of 15/25,4/17 of the PCOS group.The control group pregnancy rate higher than the PCOS group was statistically significant(P <0.05).There was no significant difference in E2,P,oocyte number and fertilization rate between two groups(P> 0.05).Pearson correlation was used to analyze the correlation between the expression of Beclin1 m RNA and BMI,P and fertilization rate in PCOS group.The Spearman rank correlation was used to analyze the correlation between the expression of Beclin1 m RNA and LH,E2,oocyte number in PCOS group.The correlation coefficient between the expression of Beclin1 m RNA and BMI,oocyte number in the PCOS group was ρ = 0.332,r = 0.310,respectively(P >0.05),which had a positive correlation with no statistically significant.There was no correlation between the expression of Beclin1 m RNA and other clinical data in PCOS group.The expression of Beclin1 m RNA and Atg5 m RNA in 19 cases of gestational and 23 cases of non-pregnancy were compared.The expression of Beclin1 m RNA in the non-pregnant group was higher than that in the non-pregnant group,and the difference was statistically significant(P <0.05).Conclusion:1 Beclin1 m RNA and Atg5 m RNA were expressed in both groups,suggesting that autophagy was prevalent in Guanqiu granulosa cells.2 Beclin1 m RNA expression between the two groups was statistically significant(P <0.05),and the expression of Beclin1 m RNA in the polycystic ovary syndrome group was higher than that in the control group.It is suggested that the autophagic activity of polycystic ovary syndrome group is higher than that of the control group,suggesting that granulosa cells can enhance the autophagic activity,confront and repair the damage caused by changes in ovarian microenvironment.3 Beclin1 and Atg5 play an important role in the initiation and extension of autophagy.The expression of Beclin1 m RNA was statistically significant(P <0.05).There was no significant difference in the expression of Atg5 m RNA between the two groups(P> 0.05).Indicating that PCOS patients with coronoid granulocyte autophagic activity changes and processes that still require large samples and further support for further studies.4 The increase of autophagic activity of granulosa cells in polycystic ovary syndrome group was not closely related to the changes of BMI and serum LH,E2 and P in triggering day.The above clinical data were not the main factors that affected the changes of autophagy activity in Guanshan granulosa cells.Nor does it affect the pregnancy rate of the group by affecting the autophagic activity of the coronoid granule cells.5 There was a difference in autophagic activity between pregnancy and non-pregnancy,and the autophagy activity was higher in non-pregnancy group.We still need to carry out more clinical data analysis,to find the reasons for the differences in autophagic activity.6 PCOS patients with hyperandrogenism,blood glucose,blood lipids,fasting insulin,whether there is smoke exposure,etc.will affect the coronoid granulocyte autophagy process.And whether it will change the crown chord granule cell autophagy affect the quality of the egg and then affect the pregnancy rate still need further discussion and validation.7 Whether there is a difference between PCOS and PCO in autophagy of coronoid granules,also need to further explore.8 This article discusses the relationship between granulocyte autophagy and polycystic ovary syndrome,providing a new clue for the study of molecular mechanism underlying the development and progression of polycystic ovary syndrome.But there are still many issues to be studied. |
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