- 學位論文
薑黃素對小鼠胚胎發育的影響
Effect of Curcumin on Mouse Embryonic Development in Vitro and in Vivo
摘要
薑黃素（curcumin）具有廣泛的藥理作用，包括抗癌、抗發炎、抗過敏及抗氧化等作用。本實驗以體外與體內兩方面探討，薑黃素對小鼠囊胚細胞的細胞毒性。本研究發現，以薑黃素與囊胚細胞共培養6小時後，僅與24μM薑黃素共培養的囊胚細胞有較高的Bax表現量；以薑黃素與囊胚細胞共培養9小時後，僅以24μM薑黃素共培養的囊胚細胞有粒線體膜電位消失的現象。另外利用免疫染色分析，發現囊胚細胞與薑黃素共培養18小時後，僅有與24μM薑黃素共培養的囊胚細胞可被偵測到較高的caspase 3活化的情況。此外以薑黃素與囊胚細胞共培養24小時後發現，6-24 μM薑黃素處理組的囊胚腔較對照組小、囊胚平均細胞數明顯減少以及抑制囊胚體外發育的能力。利用 TUNEL（terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling）進行凋亡分析，以24 μM 薑黃素與囊胚共培養24小時後，發現凋亡細胞數明顯增加。接著，利用動物模式以腹腔注射薑黃素（0，100 與 300 mg/kg/day）後並取其胚胎，發現胚胎發育至囊胚的比例，隨注射劑量增加而減少。有趣的是將取得的囊胚進行細胞凋亡與體外發育分析，發現處理組與對照組並無明顯的差異。但在體外發育的部份發現進行腹腔注射組 (100 與 300 mg/kg/day) 滋養層細胞的貼附面積較對照組小。綜合以上結果得知以薑黃素與小鼠囊胚共培養後，會造成小鼠囊胚的細胞凋亡、細胞數減少並且延緩囊胚的發育。
並列摘要
Curcumin exhibits a wide range of pharmacological effects, including anti-cancer, anti-inflammatory, anti-allergy and anti-oxidative properties. In this study, we examine the cytotoxic effects of curcumin on the blastocyst stage of mouse embryos in vitro and in vivo. Our study found that Bax protein expression levels were increase in 24 μM curcumin-treated blastocysts for 6 h. In addition, loss of mitochondrial membrane potential was measured in 24 μM curcumin-treated blastocysts for 9 h. Using immunostaining assay, we found that activation of caspase-3 was detected in 24 μM curcumin-treated blastocysts for 18 h. Moreover, that blastococle was cataplasia in 6-24 μM curcumin-treated blastocysts for 24 h. In addition, blastocysts treated with curcumin showed significant decreases in total cell number. Curcumin also inhibited postimplantation embryonic development and fewer curcumin-pretreated blastocysts reached the later stages of development than untreated control group. Furthermore, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay revealed that blastocysts treated with 24 μM curcumin showed increased apoptosis versus untreated controls. Finally, an in vivo model showed that intraperitoneal injection of curcumin（0, 100 and 300 mg/kg/day）into mice abdominal cavity caused decrease the embryo development to reach blastocyst stage following the dose-dependent manner. Interesting, studies also found that cell apoptosis and in vitro development of blastocysts had no significant differences in curcumin-injected and untreated control groups. However, the trophoblast cell attachment area of in vitro development in curcumin-injected group embryo was significant smaller than untreated control group. Our results collectively indicate that curcumin treatment of mouse blastocysts induces apoptosis, decreases cell numbers, and retards blastocyst development.