(3.234.221.162) 您好!臺灣時間:2021/04/14 15:31
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:楊爵閣
研究生(外文):Yaung, Jauyer-Ger
論文名稱:糖皮質素對豬卵母細胞體外成熟及成熟後受精能力之影響
論文名稱(外文):Effect of glucocirticoids on maturation of pig oocytes in vitro! and on their subsequent fertili{iog capacity in vitro
指導教授:李碧雪
指導教授(外文):Li Pi-Hsueh Shirley
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生理學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:1996
畢業學年度:84
語文別:中文
論文頁數:72
中文關鍵詞:糖皮質素豬卵母勝胞受精能力
外文關鍵詞:glucocorticoidpig oocyuefertilizing ability
相關次數:
  • 被引用被引用:1
  • 點閱點閱:141
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
本論文主要在探討糖皮質素對豬卵母細胞體外成熟及成熟後卵母細胞
受精能力的影響。由屠宰場取回成熟前母豬卵巢,收集卵丘卵母細胞複合
體(Cumulus-oocyte complexes),以Waymouth MB 752/1 medium 添加50
ug/ml丙酮酸鈉(Sodium pyruvate), 0.5 ug/ml黃體促素 (Luteinizing
hormone; LH), 0.5 ug/ml 濾泡促素 (Follicle-stimulating hormone;
FSH)及1 ug/ml 雌二醇(17 b-estradiol)在地塞美松(Dexamethasone;
DEX),皮質醇(cortisol), 或脫氫異雄酮(Dehydroepiandro-sterone;
DHEA)有無添加的狀態下培養24小時;然後以不添加荷爾蒙的培養液,在
DEX、 皮質醇、或DHEA有無添加的狀態下再培養16-24小時。 之後評估
受精前卵母細胞生發泡破裂(Germinal vesicle breakdown; GVB)的百分
率與卵母細胞內麩氨基硫(Glutathione; GSH)濃度變化。卵母細胞細胞質
的成熟則以體外受精後雄原核生成的百分率加以判斷。在前述添加荷爾蒙
的培養液中培養48小時後大部份(78%)的卵母細胞達到產生GVB的階段。以
漸增濃度的DEX(1-50 ug/ml)、皮質醇(0.1-10 ug/ml),或DHEA(0.1-10
ug/ml)處理卵母細胞48小時後,GVB產生的百分率隨劑量增加而被抑制。
將卵母細胞暴露在DEX (10 ug/ml)12-48小時,不同的時間會有依時間增
加而出現的GVB百分率隨暴露時間增加被抑制的百分率亦增加被抑制現象
,在12小時即達統計上顯著差異。而在卵母細胞進行培養、12、24、或36
小時之後再給予DEX(10 ug/ml)刺激至48小時,亦會有依時間增加而出現
的GVB百分率被抑制現象,在12小時即達統計上顯著差異。當卵母細胞暴
露在DEX(10 ug/ml)12小時以上,卵母細胞的成熟率即降低。降低的程度
是取決於DEX的作用,且DEX的濃度須高於(1 ug/ml)。DEX對卵母細胞成熟
的抑制作用可利用糖皮質素受體的拮抗劑RU 486加以消除。將卵母細胞暴
露在DEX中40小時,不影響精子的穿透, 多精入卵或降低卵母細胞生成雄
原核的能力。卵母細胞內GSH的濃度是影響雄原核生成的重要因子,以酵素
循環反應分析法 (Enzymeatic cycling assay)檢測出卵母細胞內GSH的濃
度。每一卵母細胞GSH的濃度約為4.4 mM。以DEX (0.01-10 ug/ml)處理
卵母細胞,不影響卵母細胞GSH的濃度。這些結果顯示,DEX對體外培養的
豬卵母細胞,抑制減數分裂但不影響細胞質的成熟。對卵母細胞減數分裂
的抑制作用並不是藉由改變卵母細胞內GSH的濃度而造成。
The present study examined the effects of glucocorticoids on
maturation ofpig oocytes in vitro and on their subsequent
fertilizing capacity in vitro.Pigcumulus-oocyte complexes
collected from prepubertal gilts at a local slaughterhouse were
cultured in Waymouth MB 752/1 medium supplemented with
sodiumpyruvate (50 ug/ml), luteinizing hormone (0.5 ug/ml),
follicle-stimulatinghormone (0.5 ug/ml), and 17b-estradiol (1
ug/ml) in the presence or absence ofdexamethasone (DEX),
cortisol, or dehrdroepiandrosterone (DHEA) for 24 h; theythen
were cultured without hormonal supplements in the presence or
absence ofDEX, cortisol, or DHEA for an additional 16-24 h.
The breakdown of germinalvesicle (GVB) and changes in
glutathione content before in vitro fertilizationwere assessed.
Cytoplasmic maturation as determined by male
pronuclearformation following fertilization in vitro was
also examined. Most oocytes(78%) achieved GVB after 48 h of
culture in medium with hormonal supplements.Treatment of
ooytes with increasing concentrations of DEX (1-50 ug/ml),
cortisol (0.1-10 ug/ml), or DHEA (0.1-10 ug/ml) for 48 h
resulted in a dose-response inhibition of GVB. Increasing
duration (12-48 h) of treatment withDEX (10 ug/ml) led to a
time-dependent inhibition of GVB, achieving statisti-cal
significance by 12 h. The addition of DEX (10 ug/ml) to
maturation mediumimmediately after culture, 12 h, 24 h, or 36
h after culture also decreasedthe percentage of oocytes with
GVB. When oocytes were exposed to DEX (10 ug/ml) for longer
than 12 h, maturation rate was reduced. The degree of
thisreduction was dependent on DEX, and aconcentration of DEX
higher than 1 ug/mlwas needed. The inhibitory effect of DEX on
the maturation of oocytes was pre-vented by the glucocorticoid
receptorantagonist RU 486. Exposure of oocytes toDEX for 40 h
did not prevent sperm penetration, affect the incidence of
poly-spermy, or decrease the ability ofoocytes to form a male
pronucleus. The con-centration of glutathione, an important
factor for male pronuclear formation,in pig oocytes was
determined by an enzymatic cycling assay. The concentrationfound
was 4.4 mM per oocyte. Exposure of oocytes to DEX (0.01-10
ug/ml) hadno effect on glutathione concentration. These
results demonstrate that DEXdirectly inhibits the meiotic but
not cytoplasmic maturation of pig oocytes invitro. This
inhibitory effect is not mediated through a decrease in the
levelof intracellular glutathione.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔