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[补肾调经方对体外培养小鼠卵母细胞中BMPR I/ALK6-Smads表达的调控]

[Bushen Tiaojing Recipe Regulated Expressions of BMPR I[/ALK6-Smads in Mouse Oocytes Cul- ture in vitro].

作者信息

Sun Xiao-Huan, Fan Li-Jie, Li Li, Wei Xue-Cong, Liu Ya-Hua, Geng Dan-Dan, Gu Xiang-Xiang, Du Hui-Lan

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2016 Oct;36(10):1241-1246.

Abstract

Objective To observe the effects of Bushen Tiaojing Recipe (BTR) on the counts of survival preantral follicles and the bone morphogenetic protein receptor II (BMPR II )/activin receptor- like kinase 6-drosophila mothers against decapentaplegic proteins (ALK6-Smads) signal pathway in oocytes cultured in vitro, and to study its mechanism for improving the quality of oocytes. Methods Prean- tral follicles were mechanically isolated from 65 female 12-day old healthy Kunming mice, which were inoculated by normal rats' serum (as the control group) , high, medium, low dose BTR containing serums (as Shen-supplementing groups) , high dose BTR containing serum + K02288 (as the inhibitor group) , respectively. All were cultured by common method in vitro. On the 6th day the counts of survival preantral follicles were compared between each Shen-supplementing group and the control group respectively. mR- NA expressions of BMPR II, ALK6, Smad1 , Smad5, and Smad8 were detected by Real-time fluorescence quantitative PCR. The protein expressions of indices mentioned above and phospho-Smadl/5/8 (p- Smadl/5/8) were detected by cellular immunofluorescence test. Results Compared with the control group, the quantity of survival preantral follicles increased in the high dose BTR containing serum group; mRNA expressions of BMPR II, ALK6, Smad5, and Smad8 were elevated, protein expressions of indi- ces mentioned above and p-Smadl/5/8 were increased in the 3 Shen-supplementing groups (P <0. 05) ; mRNA and protein expressions of Smad1 were increased in high and medium dose BTR containing serum groups (P<0.05). Compared with the high dose BTR containing serum group, protein expressions of Smad1/5/8 were reduced in the inhibitor group (P <0.05). Conclusion BTR could elevate the quantity of survival preantral follicles cultured in vitroand improve the quality of oocytes, which might be possibly as- sociated to regulating the BMPR II/ALK6-Smads signal pathway in oocytes.

摘要

目的 观察补肾调经方(BTR)对体外培养卵母细胞中存活的腔前卵泡数量及骨形态发生蛋白受体Ⅱ(BMPRⅡ)/激活素受体样激酶6-果蝇抗五聚体蛋白(ALK6-Smads)信号通路的影响,探讨其改善卵母细胞质量的机制。方法 从65只12日龄健康雌性昆明小鼠中机械分离出腔前卵泡,分别接种正常大鼠血清(作为对照组)、含高、中、低剂量BTR的血清(作为补肾组)、含高剂量BTR的血清+K02288(作为抑制剂组)。所有均采用常规方法进行体外培养。第6天分别比较各补肾组与对照组存活的腔前卵泡数量。采用实时荧光定量PCR检测BMPRⅡ、ALK6、Smad1、Smad5和Smad8的mRNA表达。采用细胞免疫荧光试验检测上述指标及磷酸化Smad1/5/8(p-Smad1/5/8)的蛋白表达。结果 与对照组相比,含高剂量BTR血清组存活的腔前卵泡数量增加;3个补肾组中BMPRⅡ、ALK6、Smad5和Smad8的mRNA表达升高,上述指标及p-Smad1/5/8的蛋白表达增加(P<0.05);含高、中剂量BTR血清组中Smad1的mRNA和蛋白表达增加(P<0.05)。与含高剂量BTR血清组相比,抑制剂组中Smad1/5/8的蛋白表达降低(P<0.05)。结论 BTR可提高体外培养的存活腔前卵泡数量,改善卵母细胞质量,其机制可能与调节卵母细胞中的BMPRⅡ/ALK6-Smads信号通路有关。

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