苦參鹼對小鼠著床前胚胎發育的影響

Effects of matrine on mouse pre-implantation embryo development

曾于烜(Yu-Hsuan Tseng)

指導教授 ： 詹文雄

中原大學/理學院/奈米科技碩士學位學程/碩士(2018年)

https://doi.org/10.6840/cycu201800069

摘要

苦參鹼是屬於豆科植物苦參的乾燥根提取出來的，許多研究已發現苦參鹼具有許多生理和藥理功效，例如具有抗心律不整、抑制癌細胞增生、抗發炎作用等功效。而研究也指出苦參鹼有能夠調節體內白血球低下的功能，並發現其具有抑制癌細胞的增殖和誘發癌細胞的凋亡功能。先前本實驗室的研究發現苦參鹼與小鼠囊胚共培養會抑制胚胎細胞增生，促使細胞凋亡以及造成甲基化的改變。苦參鹼會影響小鼠囊胚細胞H19 DNA 甲基化上升也會使 IGF-2 基因高度表現，而影響後續的胚胎發育。由先前的研究證實苦參鹼具有抑制細胞的增殖和誘導細胞凋亡的功能，根據這些結果，本研究想進一步探討苦參鹼是否會影響受精後的早期胚胎之發育，包括誘發細胞凋亡、影響胚胎細胞增生和對胚胎體外發育有所影響。本研究探討不同濃度的苦參鹼 ( 0、12.5、25、50 μM )與小鼠二細胞期胚胎共培養，探討其對早期胚胎發育的影響。利用胚胎細胞凋亡 ( TUNEL assay )、胚胎細胞增生（Hoechst assay）和胚胎體外發育（In vitro development）等分析法，發現 ( 12.5-50 μM ) 苦參鹼的具有抑制胚胎細胞增生，傷害早期胚胎發育的效應。本研究結果證實隨著苦參鹼共培養濃度的增加，胚胎細胞數目隨之的減少，進而影響早期胚胎的發育。然而，苦參鹼在早期胚胎發育中詳細的調控機制仍不清楚，需未來更進一步深入的研究。

關鍵字

苦參鹼 ； 早期胚胎 ； 胚胎發育 ； 細胞凋亡 ； 細胞增生

並列摘要

Matrine is a legume belonging to the dry roots of legumes extracted, in many studies found that matrine has many effects, such as: anti-arrhythmia, inhibition of cancer cell proliferation, anti-inflammatory effects and other effects. Many studies also found that matrine can regulate the function of leukopenia in the body, and also found that cound inhibit the proliferation and induce apoptosis in many cancer cell lines. Previous studies in our laboratory have found that co-culture of blastocysts with matrine inhibits the proliferation of embryonic cells, promotes apoptosis and affects DNA methylation. Study found that matrine increases H19 DNA methylation and induces high expression of IGF-2 gene, resulting in organ abnormalities and other effects. According the functional effects of matrine on inhibition of cell proliferation, induction of cell apoptosis, we further investigate the effects of matrin on early stage embryo development, including cell apoptosis, proliferation and in vitro embryonic development. The main purpose of this study was to investigate the effects of different concentrations of matrine (0,　12.5,　25,　50 μM) on the development of early stage embryos. Using the TUNEL assay, Hoechst staining analysis and in vitro development, studies found that the 12.5-50 μM matrin could inhibit cell proliferation and early embryonic development. Taken together, our study clearly point out that matrine has injury effects on cell proliferation and induce cell apoptosis, which can trigger developmental damage and cause embryo development injury. However, the detail regulatory mechanisms of matrin on early stage embryo development are unclear and need further investigation.

並列關鍵字

Matrine ； Early stage embryo ； Embryo development ； Apoptosis ； Cell proliferation

參考文獻

53. 林資哲, 夏奇鈮, 陳威臣, & 葉茂生. (2010). 藥用植物苦參微體繁殖及瓶苗馴化研究. 作物, 環境與生物資訊, 7 (3):, 145-152.

1. Arends, M. J., & Wyllie, A. H. (1991). Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol, 32, 223-254.

3. Barres, B. A., Hart, I. K., Coles, H. S., Burne, J. F., Voyvodic, J. T., Richardson, W. D., & Raff, M. C. (1992). Cell death in the oligodendrocyte lineage. J Neurobiol, 23(9), 1221-1230. doi:10.1002/neu.480230912

4. Beck, F., Erler, T., Russell, A., & James, R. (1995). Expression of Cdx-2 in the mouse embryo and placenta: possible role in patterning of the extra-embryonic membranes. Dev Dyn, 204(3), 219-227. doi:10.1002/aja.1002040302

5. Binder, N. K., Evans, J., Gardner, D. K., Salamonsen, L. A., & Hannan, N. J. (2014). Endometrial signals improve embryo outcome: functional role of vascular endothelial growth factor isoforms on embryo development and implantation in mice. Human Reproduction, 29(10), 2278-2286.