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標題: | 黃芩對不同品系小鼠肝細胞與庫弗氏細胞交互作用之影響 Effects of Scutellaria baicalensis Georgi on hepatocyte and Kupffer cell in different strains mice |
作者: | Wen-Chi Chang 張文綺 |
指導教授: | 吳應寧副教授(Ying-Ling Wu) |
共同指導教授: | 邱仁輝教授(Jen-Hwey Chiu) |
關鍵字: | 黃芩,肝細胞,庫弗氏細胞,小鼠品系, SbG,hepatocyte,kupffer cell,mice strain, |
出版年 : | 2007 |
學位: | 碩士 |
摘要: | 中草藥黃芩 (Scutellaria baicalensis Georgi) 在傳統醫療的應用上常被使用於肝病治療的藥方之一。過去研究發現黃芩萃取物會影響肝臟細胞的生長調控。因此,本研究目的在探討黃芩萃取物對不同品系如C57BL/6J與C3H/HeN小鼠肝細胞之作用,且建立了一個肝細胞與庫弗氏細胞交互作用的模型,來探討Toll-like receptor 4 (TLR4)突變與HBx基因轉殖小鼠中,黃芩及不同的細胞激素在此交互作用模型中的影響。
研究方法以黃芩萃取物分別處理肝細胞與庫弗氏細胞並利用MTT與trypan blue exclusion assay觀察細胞生長情形。也藉由免疫染色來了解其作用可能的路徑;在肝細胞與庫弗氏細胞共同培養模式中,使用ELISA分析細胞激素的表現。另外,我們利用肝細胞與庫弗氏細胞交叉培養模式,在不同小鼠品系中,加入黃芩萃取物與細胞激素來了解其相互作用關係。實驗結果發現,黃芩萃取物在3 mg/ml~30 mg/ml的劑量下對C57BL/6J與C3H/HeN品系小鼠之肝細胞有時間及劑量效應地促進生長作用;而在TLR4 mutant小鼠肝細胞則有降低生長之現象。當肝細胞與庫弗氏細胞共同培養於30 mg/ml的黃芩萃取物時,初代庫弗氏細胞會抑制經由黃芩引起的肝細胞生長,其機轉可能是透過TGF-β1的分泌。在5 ng/ml的TNF-α和5 ng/ml 的IL-6協同作用下會促進wild-type品系小鼠初代肝細胞的生長;而在Hbx 基因轉殖小鼠肝細胞中,無法看到促進肝細胞生長之現象。 因此,本研究結論認為黃芩萃取物對不同品系如C57BL/6J與C3H/HeN之促進肝細胞生長沒有差異。但是對於TLR4 wild-type與mutant小鼠之肝細胞生長有差異,TLR4在黃芩促進肝細胞生長中扮演了部分的角色。在肝細胞與庫弗氏細胞共同培養方面,黃芩萃取物30 mg/ml濃度下會促使庫弗氏細胞生長,並分泌TGF-β1來抑制肝細胞的生長。而庫弗氏細胞對肝細胞的抑制生長作用與TLR4無相關。在細胞激素的影響方面,細胞激素TNF-α和IL-6可以促進肝細胞生長,但並不透過TLR4的路徑。 Scutellaria baicalensis Georgi (SbG) is a well-known herbal medicine commonly used for the treatment of human liver disease. Previous investigation showed that SbG affected the proliferation of the liver cell. The aim of this study was to elucidate the effects of SbG on hepatocyte growth in different strains mice, such as C57BL/6J and C3H/HeN, and to establish a hepatocyte-Kupffer cell interaction model for analyzing the influence of SbG ,TNF-αand IL-6 in the Toll-like receptor 4 (TLR4)-mutant and HBx-transgenic mice. The effects of SbG, TNF-α and IL-6 on hepatocyte proliferation in different strains mice were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide and trypan blue exclusion assay. Cytokine production and antibody-neutralization studies were used on SbG-treated hepatocyte-Kupffer cell interaction model. The results showed that SbG stimulated the proliferation of the primary hepatocytes in a dose-dependent 3 mg/ml~30 mg/ml manner. However, the TLR4 mutant mice have less cell growth-stimulation effects. The SbG-stimulated proliferation was inhibited by TGF-β1 secretion from primary Kupffer cells. The synergistic effect of TNF-α and IL-6 induced the hepatocyte growth in the wild-type mice;whereas such effect was negative in the HBx-transgenic mice. It was concluded that there was no significant difference in terms of SbG-induced hepatocyte proliferation between different strains mice, such as C57BL/6J and C3H/HeN. However, it has significant difference between TLR4 wild-type and TLR4 mutant mice. Hence, TLR4 receptor might play a partial role in the SbG-induced hepatocyte proliferation. In the coculture model, TGF-β1 inhibit the SbG-induced hepatocyte proliferation at the dose of 30 mg/ml. TLR4 pathway was not involved in TNF-α- and IL-6-stimulated hepatocyte proliferation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/29801 |
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顯示於系所單位: | 獸醫學系 |
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