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研究生:呂光洲
研究生(外文):Leu, Guang-Chou
論文名稱:百日咳毒素對3T3-L1脂肪細胞分化過程中代謝反應的影響
論文名稱(外文):Effects of pertussis toxin on metabolic responses in the process of differentiation of 3T3-L1 adipocytes
指導教授:馮濟敏
指導教授(外文):Jim Fong
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:1994
畢業學年度:82
語文別:中文
中文關鍵詞:生物化學生物學科學百日咳脂肪細胞
外文關鍵詞:BIOCHEMISTRYBIOLOGYSCIENCE
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完全分化的小鼠3T3-L1細胞株,在經由分化促進因子(Dexame thason及IBMX)處理後的7~10天間便會有大量的脂質堆積。經由脂解刺激因子Isoproterenel刺激增加細胞由cAMP量及胰島素刺激的2-DG uptake以及CPDH活性變化的情形來加以判斷脂肪細胞分化的程度。
百日咳毒素能催化Gi及GO蛋白α-次單元體的ADP-ribosy1ation,而抑制此二蛋白的功能,由於G蛋白質在生物體中扮演著在生理上多樣化的角色,使得人們相信在分化上,G蛋白應該是相當重要的,因此利用百日咳毒素的作用及以3T3-L1脂肪細胞為模式,用來探討百日咳毒素受質的G-蛋白對此細胞分化上的影響。
實驗的結果顯示,飽滿聚集(confulence)的3T3-L1脂肪細胞以百日咳毒素處理2天後再行分化,其GPDH活性於分化第4天(day4)就有明顯地增加且於第8天此酵素活性比控制組細胞提早完成(圖二),而脂解作用、細胞內cAMP的量、及分化過程中的2-DG uptake的變化在分化前期(day2),及其值均比控制組為高,而在分化後期(day8)則得到相反的情形,因此吾人認為3T3-L1細胞的分化過程中存在著一轉換點,使得細胞在分化前期與分化後期有著完全不同的代謝反應。由此實驗的結果顯示,3T3-L1脂肪細胞的分化過程中存在著有一轉換點,在轉換點之前,百日咳毒素能影響其代謝反應,即Gi/Go蛋白在3T3-L1細胞的此外,百日咳毒素的前處理也有能促使3T3-L1脂肪細胞提早完成分化的現象。
The effect of the pertussis toxin on the differentiation of 3T3-L1 adipocytes was tested. Various dose of pertussis toxin were added to the culture dishes containing confluent preadipocytes 2 days before the addition of dexamethasone and 3-isobutyl-l-methylxanthine which induced the differentiation of the preadipocytes into adipocytes. The total cellular protein in the dishes were decreased in the presence of the differentiating agents dose-dependently by the treatment of pertussis toxin; however, after replacing the differentiation medium with the normal culture medium, the cellular protein content recovered to nearly the control level. Both the lypolytic rate and the activity of glycerol-3-phosphate dehydrogenase, a marker enzyme for differentiation of adipocyte, were enhanced in cells treated with pertussis toxin. In addition, at early stage of differentiation process (day 2), both the cAMP production and glucose transport activity have the similar phenomenon, which wer increased in dose-dependently by the pretreatment of pertussis toxin not only basal state but hormone stimulation, while at late stage of differentiation (day 8), in the fully differentiated adipocytes, the cAMP production and glucose transport activity were dose dependently decreased by pretreatment of the cells with pertussis toxin not noly basal but also hormone stimulation. However, the intracellular cAMP and glucose transport activity were more positive correlation, which analyzed by 2nd regression, the content of Giα subunit in the membranes of the processes of 3T3-L1 cell differentiation as monitored by PT-catalyzed ADP-ribosylation, which was no difference between differentiation and non-differentiation cells, were decreased in the differentiating process. On the other hand, the pertussis toxin treatment at day 2 and day 3 after initiation of differentiation, which results were to indicate that the 3T3-L1 cells differentiating process may containing a conversio point. For these results, suggestion that pretreatment of confluent p eadipocytes by pertussis toxin may enhance the differentiating rate during the subsequent differentiatiation and also may change metabolic characteristic of differentiation adipocytes.



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