湖南中医药大学学报2026，Vol.46Issue(1)：42-50,9.DOI:10.3969/j.issn.1674-070X.2026.01.006
丹参酮ⅡA对玻璃化冷冻小鼠卵母细胞保护机制的研究
Protective mechanism of Tanshinone ⅡA in vitrified mouse oocytes
摘要
Abstract
Objective To investigate the protective mechanism of Tanshinone ⅡA against vitrification-induced injury in mouse oocytes.Methods Female ICR mice aged 4-8 weeks(SPF grade)were intraperitoneally injected with 5 IU of pregnant mare serum gonadotropin(PMSG).After 48 hours,morphologically normal immature oocytes at the germinal vesicle(GV)stage were collected from the ovaries.The GV-stage oocytes were divided into control group and Tanshinone ⅡA group and cultured overnight for 16-18 hours to reach the metaphaseⅡ(MⅡ)stage.Morphologically normal MⅡ oocytes were then subjected to in vitro fertilization,and fertilization,cleavage,and blastocyst formation were observed in each group.The protein expression level of aquaporin 3(AQP3)on the cell membrane was detected in MⅡ oocytes from the control group and the Tanshinone ⅡA group using immunofluorescence technique.In another experiment,GV-stage oocytes were divided into four groups for culture until maturation:a control group(non-vitrified),a Tanshinone ⅡA group(non-vitrified),a control group(vitrified),and a Tanshinone ⅡA group(vitrified).Fertilization rate,cleavage rate,and blastocyst formation rate were observed and analyzed in the four groups.A cryomicroscope was employed to observe the effect of TanshinoneⅡA on intracellular ice formation(IIF)during the cryopreservation of MⅡ oocytes.Results Compared with the control group,the Tanshinone ⅡA group showed increased rates of oocyte maturation,fertilization,cleavage,and blastocyst formation(P<0.05),with the highest blastocyst rate observed when treated with 0.5 μg/mL Tanshinone ⅡA.The protein expression level of AQP3 was significantly higher in the Tanshinone ⅡA group than in the control group(P<0.000 1).Compared with the control group(non-vitrified),the TanshinoneⅡA group(non-vitrified)showed significantly higher fertilization,cleavage,and blastocyst formation rates(P<0.05).Similarly,compared with the control group(vitrified),the Tanshinone ⅡA group(vitrified)also exhibited significantly increased fertilization,cleavage,and blastocyst formation rates(P<0.05).Additionally,IIF occurred at a lower temperature with a more concentrated temperature range,resulting in a lower probability of ice formation during cooling.Conclusion TanshinoneⅡA improves the survival rate and developmental potential of vitrified oocytes by upregulating the expression of AQP3 on the oocyte membrane and reducing the temperature of IIF during cryopreservation.关键词
卵母细胞/玻璃化冷冻/丹参酮ⅡA/水通道蛋白3/细胞内冰晶/渗透性Key words
oocyte/vitrification/Tanshinone ⅡA/AQP3/intracellular ice crystals/permeability分类
医药卫生引用本文复制引用
王婕,刘志成,张彩凤,郝焱,游卉,尤昭玲,王慧颖,金波..丹参酮ⅡA对玻璃化冷冻小鼠卵母细胞保护机制的研究[J].湖南中医药大学学报,2026,46(1):42-50,9.基金项目
育龙计划-三龙人才项目(2022-BUCMSZYLRC05) （2022-BUCMSZYLRC05）
深圳市龙岗区科技创新专项(LGWJ2024-37) （LGWJ2024-37）
深圳市"医疗卫生三名工程"项目(SZZYSM202311019) （SZZYSM202311019）
北京中医药大学深圳医院(龙岗)联合银丰低温医学科技有限公司合作课题(KJK-HX-2023-001) （龙岗）
国家中医药管理局"尤昭玲全国名中医传承工作室"建设项目(国中医药办人教函[2022]5号). （国中医药办人教函[2022]5号）
