Study of effect and mechanism of berberine on cumulus expansion in mice with polycystic ovary syndrome
Objective To investigate the effect and mechanism of berberine(Ber)on cumulus expansion in mice with polycystic ovary syndrome.Methods A total of 28 SPF grade female C57BU6 mice aged 21 to 25 days were selected and divided into control group,PCOS group,Ber low-dose group,and Ber high-dose group according to random number table method,with seven mice in each group.Control group was injected subcutaneously with equal volume of corn oil solvent,PCOS group,Ber low-dose group,and Ber high-dose group were injected subcutaneously with dehydroepiandrosterone(6 mg/100 g,soluble in corn oil solvent)for 21 consecutive days.The success criterion of PCOS model construction was based on the disturbance of estrous cycle in mice.Then for 28 consecutive days,Ber low-dose group was given 200 mg/kg Ber and Ber high-dose group was given 400 mg/kg Ber,PCOS group was given equal dose of normal saline.Pregnant horse serum gonadotropin and human chorionic gonadotrop in were intraperitoneally injected into each group of mice,and cumulus oocyte complex(COC)was obtained from the ampulla of the fallopian tube,and cultured in vitro for 0 and 6 h to observe the expansion of cumulus.The expression level of FTO protein in cumulus gran-ulosa cells in each group was detected by immunohistochemistry.PC3.1 plasmid(PC3.1 group)and PC3.1-FTO plasmid(PC3.1-FTO group)were transfected into human granulosa cell line(KGN cells),and the expression level of hyaluronate acid synthetase 2(HAS2)was detected by im-munofluorescence.KGN cells were transfected with NC siRNA(NC group)and FTO siRNA(siFTO group),and the expression level of HAS2 in the two groups was detected by immunofluorescence.Results Compared with control group,the estrous cycle of mice in PCOS group lost periodic changes.The results indicated that the PCOS mouse model was successfully constructed.Compared with PCOS group,the estrous cycle of mice in Ber low-dose group recovered gradually,while the estrous cycle of mice in Ber high-dose group still lost its periodic change.Compared with control group,the distribution of COC cumulus cells in PCOS group was more dense and cumulus expansion was blocked.Compared with PCOS group,the distribution of COC cumulus cells in Ber low and high dose groups was more loose,and the obstruction of cumulus expansion was significantly im proved.Compared with control group,the expression level of FTO in cu-mulus granulosa cells in PCOS group was increased(P＜0.05).Compared with PCOS group,the expression level of FTO in cumulus gran-ulosa cells of Ber low and high dose groups was decreased(P＜0.05).Compared with PC3.1 group,HAS2 expression level in KGN cells in PC3.1-FTO group was decreased(P＜0.05).Compared with NC group,HAS2 expression level of KGN cells in siFTO group was increased(P＜0.05).Conclusion The Ber may induce an increase in HAS2 expression levels by reducing FTO expression levels in granulosa cells,thereby im-proving the cumulus expansion of PCOS.
Polycystic ovary syndromeFat mass and obesity-associated geneCumulus expansionHyaluronic acid synathetase 2
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维普
