芹菜素对非洲爪蟾卵母细胞体外成熟的影响及其作用机制

doi:10.3969/j.issn.1004-616x.2013.04.003

癌变·畸变·突变

芹菜素对非洲爪蟾卵母细胞体外成熟的影响及其作用机制

张伦,高舒,彭志晴,侯丽颖,吴坤

哈尔滨医科大学营养与食品卫生教研室，黑龙江哈尔滨 150081

收稿日期:2013-03-26 修回日期:2013-05-30 出版日期:2013-07-30 发布日期:2013-07-30
通讯作者: 吴坤，E-mail：wukun_15000@126.com
作者简介: 张伦 (1987- )，女，河南商城人，硕士研究生，研究方向：植物化学物的生物学作用。

Effect and mechanism of apigenin on in vitro maturation of Xenopus oocytes

ZHANG Lun，GAO Shu，PENG Zhi-qing，HOU Li-ying，WU Kun*

Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical University, Harbin 150081, Heilongjiang, China

Received:2013-03-26 Revised:2013-05-30 Online:2013-07-30 Published:2013-07-30
Contact: WU Kun，E-mail：wukun_15000@126.com

摘要/Abstract

摘要：

目的：探讨不同浓度的芹菜素对非洲爪蟾卵母细胞体外成熟的影响及作用机制。方法：将健康的非洲爪蟾卵母细胞移入含不同浓度 (0、20、80、160和500 μmol/L)的芹菜素培养液中培养，另设阳性对照 (孕酮5 μg/mL)组。每隔1 h观察生发泡破裂 (germinal vesicle breakdown，GVBD)发生情况、并记录发生GVBD的细胞数目；同时用10%的三氯乙酸固定，观察细胞核的变化；采用荧光染色法在激光共聚焦显微镜下观察细胞染色体的变化。收集上述各浓度芹菜素处理8 h后的卵母细胞，采用Western blot法检测卵母细胞成熟相关蛋白p-Erk1、Mos和CyclinB2的表达。结果：与阴性对照组相比，当芹菜素浓度为80~500 μmol/L且作用时间在4~8 h时，卵母细胞GVBD发生率均明显升高 (P均<0.05)。在80~500 μmol/L芹菜素处理细胞8 h后，p-Erk1、Mos和CyclinB2蛋白表达量明显高于阴性对照组 (P<0.05)。结论：芹菜素在卵母细胞体外培养过程中促进了第1次减数分裂的开启，其机制可能与MPF和MAPK信号通路的活化有关。

关键词: 芹菜素, 卵母细胞, 体外成熟, 非洲爪蟾

Abstract:

OBJECTIVE: To explore the effect and mechanism of different doses of apigenin on in vitro maturation of Xenopus oocytes. METHODS：Healthy Xenopus oocytes were cultured in different doses (20，80，160 and 500 μmol/L) of apigenin. Meanwhile negative and positive control groups were set. Then we observed and recorded the rate of germinal vesicle breakdown (GVBD) per hour，confirmed the transformation of cell nucleus and chromosome by means of trichloroacetic acid fixation and fluorescence staining. Finally oocytes were collected at 8 h in each group，and the expressions of p-Erk1，Mos and CyclinB2 were detected by Western blot. RESULTS：Compared with negative control group，GVBD rates were all highly increased when apigenin-treated dose were 80，160 and 500 μmol/L，and time were 4-8 h (P<0.05). The expressions of p-Erk1，Mos and CyclinB2 were all markedly increased when the dose of apigenin were between 80 and 500 μmol/L (P<0.05). CONCLUSION：Apigenin may play an important role in promoting the maturation of Xenopus oocytes in vitro through regulation of mature promoting factor (MPF) and mitogen-activated protein kinase (MAKP).

Key words: apigenin, oocytes, in vitro maturation, Xenopus

张伦,高舒,彭志晴,侯丽颖,吴坤. 芹菜素对非洲爪蟾卵母细胞体外成熟的影响及其作用机制[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004-616x.2013.04.003.

ZHANG Lun，GAO Shu，PENG Zhi-qing，HOU Li-ying，WU Kun. Effect and mechanism of apigenin on in vitro maturation of Xenopus oocytes[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.04.003.

参考文献

[1] Cotter AA，Cashman KD. The effect of two dietary and a synthetic phytoestrogen on transepithelial calcium transport in human intestinal-like Caco-2 cells[J]. Eur J Nutr，2005，44 (2)：72-78.

[2] Tsutsumi N，Kawashima K，Nagata H，et al. Effects of KCA-012 on bone metabolism in organ culture[J]. Jpn J Pharmacol， 1995，67 (2)：169-171.

[3] Ailene M，Martha S，Carmen R，et al. Effects on the growing mouse oocyte are influenced by diet[J]. Biol Reprod，2009，80 (5)：1066-1071.

[4] Vodková Z，Rajmon R，Petr J，et al. Effects of genistein and genistin on in vitro maturation of pig oocytes[J]. Czech J Anim Sci，2008，53 (1)：1-8.

[5] Chan Wen-Hsiung. Impact of genistein on maturation of mouse oocytes，fertilization，and fetal development[J]. Reprod Toxicol， 2009，28 (1)：52-58.

[6] Naoko Y，Katsushige M. Effect of physiological levels of phytoestrogens on mouse oocyte maturation in vitro[J]. Cytotechnol，2012，64 (3)：241-247.

[7] 张晓东，臧益民，谢安. 用于离子通道基因表达的爪蟾卵母细胞的分离和培养[J]. 心脏杂志，2001，13 (2)：120-122.

[8] Ghodageri MG，Katti P. In vitro induction/inhibition of germinal vesicle breakdown (GVBD) in frog (Euphlyctis cyanophlyctis) oocytes by endocrine active compounds[J]. Drug Chem Toxicol， 2013，36 (2)：217-223.

[9] Abrieu A，Doree M，Fisher D. The interplay between cyclin-B-Cdc2 kinase (MPF) and MAP kinase during maturation of oocytes[J]. J Cell Sci，2001，114 (2)：257-267.

[10] Picard A，Pahlavan G，Robert S，et al. Effect of organochlorine pesticides on maturation of starfish and mouse oocytes[J]. Toxicol Sci，2003，73 (1)：141-148.

[11] Sadler SE，Archer MR，Spellman KM. Activation of the progesterone- signaling pathway by methyl-beta-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa Galphas[J]. Dev Biol，2008，322 (1)：199-207.

[12] Bodart JF，Gutierrez DV，Nebreda AR，et al. Characterization of MPF and MAPK activities during meiotic maturation of Xenopus tropicalis oocytes[J]. Dev Biol，2002，245 (2)：348-361.

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芹菜素对非洲爪蟾卵母细胞体外成熟的影响及其作用机制

Effect and mechanism of apigenin on in vitro maturation of Xenopus oocytes

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