人參皂苷Rg-1誘導小鼠胚胎發育傷害和內源性細胞凋亡

人參皂苷Rg-1，為一皂苷類化合物富含於亞洲人參 (Panax ginseng) ，其具許多藥理作用，例如神經保護、刺激免疫反應和抗糖尿病作用等；此外其在於亞洲地區常做為懷孕婦女的營養補充品。因此，人參皂苷Rg-1在懷孕時期的作用相當受到注目。在此我們利用小鼠探討其對胚胎發育的影響，人參皂苷Rg-1的作用機制對於小鼠囊胚依然未知。首先，我們在老鼠囊胚培養液中加入25、50和100 µM的人參皂苷Rg-1，觀察其對囊胚的影響，包括TUNEL凋亡分析、胚胎細胞增生和後續發育分析。研究發現人參皂苷Rg-1會導致細胞凋亡， 100 µM人參皂苷Rg-1會促使小鼠囊胚的凋亡及抑制細胞增生；螢光染色數據也顯示人參皂苷Rg-1增加小鼠囊胚的氧化壓力產生和Bax/Bcl-2的比率。藉由預處理抗氧化劑NAC (N-Acetyl-Cysteine)可抑制ROS (Reactive oxygen species) 產生，同時防止細胞凋亡。另外，人參皂苷Rg-1會導致囊胚中粒線體膜電位喪失並且造成caspase-9和caspase-3的活化，但不會影響caspase-8。同時我們發現人參皂苷Rg-1在25µM濃度下會影響早期的囊胚發育；在100µM濃度下更會造成著床後的生長終止。綜合以上實驗我們推測在懷孕時期補充人參皂苷Rg-1可能具有潛在的危險，此外人參皂苷Rg-1會透過內源性的凋亡途徑促進小鼠囊胚細胞死亡。

關鍵字

人參皂?Rg-1 ；凋亡；胚胎細胞；發育

並列摘要

Ginsenosides-Rg1, the most abundant compound found in Asian ginseng (Panax ginseng), has demonstrated pharmacological effects such as neuroprotective, immune-stimulant and anti-diabetic agent. Pregnant women, especially in Asian community, consume the ginseng as nutritive supplement. Thus, the effects of Ginsenoside-Rg1 on mouse embryo need to be investigated. Moreover, the regulatory mechanisms of Ginsenoside-Rg1 affected mouse blastocyst remains unknown. In this study, we also explored intracellular signaling mechanisms involved in that process. First, the mouse blastocysts were exposed by 25, 50 and 100 µM Ginsenoside-Rg1. Cell viability and proliferation in blastocyst, early embryonic development (in vitro) as well as blastocyst outgrowth were evaluated by TUNEL assay, Hoechst staining and morphological observation under microscope, respectively. Fluorescence staining was performed to observe the apoptosis pathway induced by Ginsenoside-Rg1. The result showed that 100 µM Ginsenoside-Rg1 could induce cell apoptosis and inhibit cell proliferation in mouse blastocyst. Fluorescence staining data revealed that Ginsenoside-Rg1 increased ROS (Reactive oxygen species) production and of Bax/Bcl-2 ratio of mouse blastocyst. That ROS production could be inhibited by NAC (N-Acetyl-Cysteine) pretreatment which lead to cell apoptosis prevention. In addition, Ginsenoside-Rg1 caused loss of MMP (Mitochondrial Membrane Potential) and triggered the activation of caspase-9 and caspase-3, but not caspase-8. Other adverse effects of Ginsenoside-Rg1 were retardation of early embryonic development at 25 µM or more concentration and embryo outgrowth disruption after blastocyst outgrowth (in vitro) at 100 µM concentrations. Taken together, our results showed that the intake of Ginsenoside-Rg1 has potentially hazardous during pregnancy. Furthermore, Ginsenoside-Rg1 exposure promotes cell apoptosis in mouse blastocyst and it occurs through intrinsic pathway.

並列關鍵字

apoptosis ； development ； embryos ； Ginsenoside-Rg1

參考文獻

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人參皂苷Rg-1誘導小鼠胚胎發育傷害和內源性細胞凋亡

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