摘要
目的 探索小檗碱(Ber)对多囊卵巢综合征小鼠卵丘扩展的作用及机制。方法 选取育龄21~25 d的SPF级雌性C57BL/6小鼠28只,按照随机数字表法将其分为对照组、PCOS组、Ber低剂量组、Ber高剂量组,各7只。对照组皮下注射等体积玉米油溶剂,PCOS组、Ber低剂量组、Ber高剂量组均皮下注射脱氢表雄酮(6 mg/100 g,溶于玉米油溶剂),连续21 d。以小鼠出现动情周期紊乱为PCOS模型构建成功标准。随后连续28 d,Ber低剂量组灌胃200 mg/kg Ber,Ber高剂量组灌胃400 mg/kg Ber,PCOS组灌胃等剂量的生理盐水。向各组小鼠腹腔注射孕马血清促性腺激素及人绒毛膜促性腺激素,于输卵管壶腹部获取卵丘卵母细胞复合物(COC),体外培养0、6 h观察小鼠卵丘扩展情况。免疫组织化学检测各组小鼠卵丘颗粒细胞中FTO蛋白表达水平。向人颗粒细胞瘤样细胞系(KGN细胞)中转染PC3.1质粒(PC3.1组)及PC3.1-FTO质粒(PC3.1-FTO组),免疫荧光检测两组细胞中透明质酸合成酶2(HAS2)表达水平。向KGN细胞中转染NC si RNA(NC组)及FTO si RNA(si FTO组),免疫荧光检测两组细胞中HAS2表达水平。结果 与对照组比较,PCOS组小鼠动情周期失去周期性变化。提示PCOS小鼠模型构建成功。与PCOS组比较,Ber低剂量组小鼠动情周期逐渐恢复,Ber高剂量组小鼠动情周期仍失去周期性变化。与对照组比较,PCOS组小鼠COC卵丘细胞分布更致密,卵丘扩展受阻。与PCOS组比较,Ber低、高剂量组小鼠COC卵丘细胞分布更松散,卵丘扩展受阻明显改善。与对照组比较,PCOS组卵丘颗粒细胞中FTO表达水平升高(P<0.05)。与PCOS组比较,Ber低、高剂量组卵丘颗粒细胞中FTO表达水平均降低(P<0.05)。与PC3.1组比较,PC3.1-FTO组KGN细胞中HAS2表达水平降低(P<0.05)。与NC组比较,si FTO组KGN细胞中HAS2表达水平升高(P<0.05)。结论 Ber可能通过降低卵丘颗粒细胞中FTO表达水平,诱发HAS2表达水平升高,从而改善PCOS卵丘
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 C57BL/6 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 granulosa 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 immunofluorescence.KGN cells were transfected with NC si RNA(NC group) and FTO si RNA(si FTO 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
作者
夏晴
王大勇
葛方亮
郑菲菲
赵雪
叶可
刘璐璐
刘雁峰
XIA Qing;WANG Dayong;GE Fangliang;ZHENG Feifei;ZHAO Xue;YE Ke;LIU Lulu;LIU Yanfeng(Department of Gynecology,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijng100700,China;Department of Biochemistry and Molecular Biology,Basic Medical College,Harbin Medical University,Heilongjiang Province,Harbin150086,China)
出处
《中国医药导报》
CAS
2024年第13期1-6,共6页
China Medical Herald
基金
中国博士后科学基金资助项目(2022M720521)
国家自然科学基金资助项目(82305298、82274567、82074480)。
