https://scholars.lib.ntu.edu.tw/handle/123456789/87860
標題: | 表皮生長因子對菜鴨卵巢粒性細胞類固醇生成調控之研究 | 作者: | 吳兩新 | 關鍵字: | 菜鴨;表皮生長因子;孕酮;Tsaiya duck;epidermal growth factor;progesterone | 公開日期: | 2005 | 出版社: | 臺北市:國立臺灣大學動物科學技術學系暨研究所 | 摘要: | 菜鴨犧牲後,取其排卵前的最大濾泡(F1)之粒性細胞層,將粒性細胞層區 分為胚盤(GD)、非胚盤(NGD)區域,以及混合的粒性細胞,利用體外培養 模式,並以酵素免疫分析法與西方墨染法分別檢測表皮生長因子(EGF)對粒性細 胞之P4 分泌量與StAR、P450scc 及3β-HSD 之表現。 結果發現,在體外培養下,GD 區粒性細胞P4 之基礎分泌量明顯(P < 0.05) 比NGD 區為低;而GD 區之P4 分泌能力在培養12 小時即可被hEGF 所抑制, NGD 區須到24 小時才有相似效果;但若外源給予oLH 刺激,則GD 區和NGD 區都在培養12 小時後被hEGF 抑制了P4 的分泌量。 在混合之粒性細胞情況下, hEGF 可抑制P4 生成之PKA 路徑,而由西方墨 染法的結果顯示,hEGF 對StAR 與P450scc 之表現具有抑制效果。添加 mitogen-activated protein kinase(MAPK kinase)抑制劑PD98059 與oLH、hEGF 共培養,發現hEGF抑制oLH引發的P4分泌量會被PD98059 所阻斷;此暗示hEGF 與粒性細胞膜上EGF 受體結合後,會活化胞內MAPK 路徑,進而影響粒性細胞 P4 生成訊息傳遞路徑。 綜合上述試驗結果:(1)EGF 會抑制粒性細胞P4 生成相關蛋白質 — StAR 及P450scc 之表現;(2)EGF 是經由胞內MAPK 路徑來影響粒性細胞之P4 生成。 The largest follicle(F1)was taken from Tsaiya duck and granulosa layer was isolated and then separated according to their proximity to GD. The cell proximal to GD, distal to GD(NGD)and mixture of both were the three varieties of granulosa cells investigated. In vitro culture of granulosa cells was used to determine the P4 levels in culture medium and StAR, P450scc and 3β-HSD protein expression in cells. Results showed that P4 produciton of granulosa cells proximal to GD is significantly lower than granulosa cells distal to GD, i.e(NGD)(P < 0.05). The basal and oLH-induced P4 production of both types of granulosa cells were inhibited by EGF. The efficiency of basal P4 production inhibited by EGF in GD group was higher than NGD group. However, the efficiency of oLH-induced P4 production inhibited by EGF was similar between two groups. EGF inhibited PKA pathway of P4 secretion when mixed granulosa cells were cultured in vitro. Western blot showed decreased expression of EGF co-cultured StAR and P450scc. The inhibitory effect of oLH-induced P4 production by EGF was disrupted by addition of MEK inhibitor(PD98059), indicating that EGF inhibited P4 production of granulosa cell via affecting MAPK pathway. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/16472 | 其他識別: | 932313B002113 | Rights: | 國立臺灣大學動物科學技術學系暨研究所 |
顯示於: | 動物科學技術學系 |
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932313B002113.pdf | 300.81 kB | Adobe PDF | 檢視/開啟 |
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