Study On The Molecular Mechanism Of Local Ovary Insulin Resistance In Polycystic Ovary Syndrome Patients With Phlegm-dampness Syndrome Via MiR-133a-3p/PI3K/AKT Signaling Pathway

Objective: To analyze the characteristic manifestations of basic endocrine,glucose metabolism,and outcome of controlled ovarian hyperstimulation and in vitro fertilization of polycystic ovary syndrome(PCOS)patients with phlegm-dampness syndrome.To study the targeting effect of miR-133a-3p on PI3K/AKT signaling pathway in ovarian granulosa cells(GCs).And to explore the regulatory role of miR-133a-3p in ovary insulin resistance(IR).Methods: Patients who underwent in vitro fertilization and embryo transfer(IVF-ET)at the Integrative Medicine Center for Reproductive and Heredity of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from November 2020 to June 2021 were selected.32 cases in the phlegm-dampness syndrome PCOS group,21 cases in the non-phlegm-dampness syndrome PCOS group and 36 cases in the control group were included,respectively.Compared and analyzed the reproductive endocrine,glucose metabolism,and controlled ovarian hyperstimulation outcome between the three groups.We did target prediction and enrichment analysis on miR-133a-3p.Human ovarian GCs were cultured in vitro,and Western Blot was used to detect the PI3K/AKT signaling pathway activity of GCs between phlegm-dampness syndrome PCOS group and other groups.miR-133a-3p mimic and miR-133a-3p inhibitor were respectively transfected into GCs.Western Blot was used to detect the changes of PI3K/AKT signaling pathway activity and the expression changes of glucose metabolism-related indicators after transfection.Results:(1)BMI,WHR,T,FPG,FINS and HOMA-IR in the phlegm-dampness syndrome PCOS group were higher than the non-phlegm-dampness syndrome PCOS group and the control group,and LH was higher than the control group(P<0.05).(2)The dosage of Gn in the phlegm-dampness syndrome PCOS group was higher than that in the non-phlegm-dampness syndrome PCOS group and the control group(P<0.05),phlegm-dampness syndrome PCOS group and non-phlegm-dampness syndrome PCOS group had more follicles >14 mm on trigger day than the control group(P<0.01).(3)The egg capture rate and high-quality embryo rate of phlegm-dampness syndrome PCOS group were lower than those of non-phlegm-dampness syndrome PCOS group and the control group(P<0.01),and the normal fertilization rate was lower than that of the control group(P<0.05).(4)The expressions of PI3 K and p-AKT proteins in the phlegm-dampness symdrome PCOS group were lower than the non-phlegm-dampness syndrome PCOS group and the control group(P<0.001).(5)With miR-133a-3p overexpressed,the expressions of PI3 K,p-AKT,and GLUT4 were reduced(P<0.05),and the expressions of p-GSK-3 β and p-FOXO1 were enhanced(P<0.001).After inhibiting the expression of miR-133a-3p,the expressions of PI3 K,p-AKT,and GLUT4 increased(P<0.05),and the expressions of p-GSK-3β and p-FOXO1decreased(P<0.05).Conclusion: PCOS patients with phlegm-dampness syndrome are more likely to develop abdominal obesity,reproductive endocrine and glucose metabolism disorders,and have poor in vitro fertilization outcome.The activity of PI3K/AKT signaling pathway in ovarian GCs of PCOS patients with phlegm-dampness syndrome is inhibited,and abnormally expressed miR-133a-3p may mediate ovary IR of PCOS patients with phlegm-dampness syndrome patients by targeting PI3K/AKT signaling pathway.