Mechanism of electroacupuncture on improving insulin resistance and IVF-ET pregnancy outcome in polycystic ovary syndrome patients with phlegm-damp syndrome based on DNA methylation
10.3760/cma.j.cn101441-20231030-00187
- VernacularTitle:基于DNA甲基化探讨电针改善痰湿型PCOS患者胰岛素抵抗及IVF-ET妊娠结局的作用机制
- Author:
Lu GUAN
1
;
Shan XIANG
;
Fang LIAN
;
Haicui WU
;
Yuan LI
Author Information
1. 山东中医药大学第一临床医学院，济南 250014
- Publication Type:Journal Article
- Keywords:
Polycystic ovary syndrome;
Electroacupuncture;
Methylation;
Phlegm dampness;
Insulin resistance
- From:
Chinese Journal of Reproduction and Contraception
2024;44(4):359-365
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the mechanism of electroacupuncture (EA) on improving insulin resistance and in vitro fertilization and embryo transfer (IVF-ET) pregnancy outcome in polycystic ovary syndrome (PCOS) patients with phlegm-damp syndrome based on DNA methylation and insulin metabolism signaling pathway. Methods:In this prospective randomized controlled study, 100 PCOS patients with phlegm-damp syndrome who underwent IVF-ET in Department of Reproduction and Genetics in Affiliated Hospital of Shandong University of Traditional Chinese Medicine from January 2020 to December 2022 were enrolled and divided into a treatment group (EA therapy) and control group (placebo needling) by random number table, 50 patients in each group. Fixed antagonist regimen was used to promote ovulation in the two groups. Patients received EA therapy and placebo needling respectively twice a week from the menstrual cycle before oocyte retrieval till human chorionic gonadotrophin injection day. The granulosa cells were collected. The improvement of phlegm and dampness syndrome, homeostasis model assessment insulin resistance (HOMA-IR), dosage and number of days of gonadotropins (Gn) used, number of oocytes retrieved and high-quality embryos, fertilization rate, clinical pregnancy rate, early miscarriage rate and live birth rate of patients in the two groups were compared. Bisulfite amplicon sequencing was used to evaluate the methylation levels of the INSR gene promoter region in ovarian granulosa cells of patients in the two groups. Real-time polymerase chain reaction and Western blotting technology were used to detect the expression of INSR, PI3K and GLUT4. Results:The phlegm-dampness score (15.23±1.57) and HOMA-IR (2.82±0.39) of the experimental group after treatment were significantly lower than those before treatment (21.65±3.61 and 3.34±0.56), and the differences were statistically significant ( P<0.001, P=0.014). The differences of the phlegm-dampness score (-5.76±2.86) and HOMA-IR (-2.67±0.06) before and after treatment in the experimental group were significantly greater than those in control group (-1.64±0.84, -0.11±0.04), and the differences were all statistically significant ( P<0.001, P=0.021). In the experimental group, the dosage of Gn used by patients [(2 119.53±338.28) U] and the duration of Gn used [(10.16±1.25) d] were significantly reduced compared with control group [(2 405.65±434.20) U, P=0.005; (10.94±1.46) d, P=0.026], and the number of high-quality embryos (3.54±1.04) was significantly increased compared with control group (2.66±1.87, P=0.014). Fertilization rate [66.91% (552/825)], clinical pregnancy rate [63.27% (31/49)] and live birth rate [51.02% (25/49)] were significantly higher in the experimental group than in control group [60.20% (475/789), 41.67% (20/48), 31.25% (15/48)], and the differences were all statistically significant ( P=0.005, P=0.033, P=0.048). There were no significant differences in the number of oocytes retrieved and early abortion rate between the two groups (all P>0.05). The methylation levels of INSR gene promoter sites 38, 47, 56, 59, 94 and 143 in the experimental group were significantly lower than those in control group (all P<0.05), but there were no significant differences in methylation levels of sites 71, 74, 154, 156 and 162 between the two groups (all P>0.05). Compared with control group, the mRNA and protein expression of INSR, PI3K and GLUT4 was up-regulated in the experimental group (all P<0.05). Conclusion:EA may improve IR by down-regulating INSR methylation levels and up-regulating INSR, PI3K and GLUT4 expression, thereby enhancing the quality of embryos and improving the pregnancy outcome in PCOS patients.
