The Relevant Research Of MiR-483-5p In Human Serum And Follicular Fluid With Polycystic Ovarian Syndrome

Objective: Detecting the different expression of miR-483-5p in serum and follicular fluid in PCOS patients and normal control group, as well as in different subtypes in PCOS patients, we discuss the correlation between the expression of miR-483-5p and obesity, hyperinsulinmia and hyperandrogenism, and to identify PCOS biological signature of miRNA, find the pathogenesis of PCOS and clinical classification based on epigenetics, elucidate the development mechanism of PCOS and provide new ideas for diagnosis and treatment.Methods: From January 2015 to December 2015, 60 infertile women underwent their IVF/ICSI-ET cycles in Second Hospital of Hebei Medical University, which were divided into 30 PCOS group and 30 normal control group. The diagnosis of PCOS was according to the definition of the Rotterdam meeting. Selection of normal ovarian function of patients with the normal menstrual cycle(28 to 30 d), basic sexual hormone and the number of bilateral ovaries sinus follicle(unilateral sinus follicle = 6-10) as the standard. Two groups of patients aged < 35 years old, and had no history of ovarian cysts, ovarian tumors, ovarian surgery, ovarian radiotherapy and ovarian chemotherapy, endometriosis, hyperthyroidism, hypothyroidism, hyperprolactinemia and other endocrine diseases. Grouping is as follows, group A: according to body mass index(BMI) of 30 patients with PCOS can be divided into: A1 group(obese), BMI equal to or more than 25 ㎏/㎡, A2 group(non-obese group), BMI less than 25 ㎏/㎡; group B: normal control group. Diagnostic criteria for obesity: according to the Asian BMI criteria. The blood was centrifuged for 5 min at 2,000×g before put in-80 ℃ refrigerator. All patients for controlled ovarian hyperstimulation treatment, when at least one follicle diameter is 17 mm or greater, given human chorionic gonadotrophin 250μg, after 36-38 hours puncture egg. At the time of oocyte retrieval, follicle fluid from the punctured follicle was collected from each follicle whose diameter is 17 mm or greater. The fluid was centrifuged for 10 min to pellet any blood and debris and reserved the supernatant. Using Trizol method to extract serum and follicular fluid total RNA and using the NanaDrop Spectrophotometer to detect the total RNA concentration, purity and integrity; After reverse transcription, the Real Time PCR reaction condition was 95℃ for 3 min, 40 cycles of 95℃ for 12 s, 62℃ for 40 s, according to SYBR ? Premix Ex TaqTM Ⅱ kit manual operation. Each sample of the PCR reaction parallel repeat three holes, all independent RT-PCR experiments repeated three times. Expression of miRNA were analyzed relative to the external constant expression of the cel-mir-39-3p in each of the individual serum and follicle fluid samples. Significance was defined at P<0.05.Results: Basic data statistical analysis shows that,compared with normal control group, PCOS group shows no statistical difference between age, duration of infertility, basal serum FSH value and basal serum E2 value(P>0.05). Basal serum LH value、basal serum T value、serum Glu value and serum INS value in PCOS group were higher than in normal control group(P<0.05).Experimental data statistical analysis shows that, compared with PCOS group and normal control group, the relative expression of miR-483-5p in serum shows no statistical difference(P > 0.05), the relative expression of miR-483-5p in follicular fluid was statistically difference(P < 0.01). PCOS obesity group compared with PCOS non-obese group, the relative expression of miR-483-5p in serum and follicular fluid has no statistical difference(P > 0.05).Conclusions: PCOS can cause cryptorrhea, insulin resistance, and increase the risk of type 2 diabetes. Trend of the expression of miR-483-5p in serum and follicular fluid are same, which shows the PCOS group is higher than the normal control group, but only the expression of miR-483-5p in the follicular fluid is statistically significant. miRNA in serum is not necessary to reflect the ovarian fundamental change, the role and importance of miRNA in the blood of the patient with PCOS is still need further research. Both in serum and follicular fluid of PCOS obesity group and PCOS non-obese group, the expression of miR-483-5p has no statistical difference. Perhaps by PCOS atypia, research object, the different control group to explain the differences between them.Therefore, more studies are needed to solve these divergent findings.