以戴奧辛活化冷光酵素基因表現法研究中藥 抑制人類肝癌細胞CYP1A1活性之影響

The Inhibition Effect for Chinese Herbs on TCDD Induction of CYP1A1 in Human Hepatoma Cells

陳中豪(Chung-Hao Chen)

指導教授 ： 王雅玢

中原大學/工學院/土木工程研究所/碩士(2011年)

https://doi.org/10.6840/cycu201100897

摘要

戴奧辛類化合物因其高脂溶性而容易透過食物鏈進入人體中，產生「假性荷爾蒙」效應，干擾內分泌系統的調控，毒害人體健康，造成肝臟毒性及致癌高風險。目前的研究多以人類乳癌細胞或老鼠肝臟細胞，探討藥物或食材萃取物如何預防或抑制戴奧辛進入人體後，代謝反應所產生之致癌代謝產物，本研究係以人類肝癌細胞株所建立之戴奧辛活化冷光酵素基因表現法 (DRE-Luciferase assay) 為測試平台，在三種不同反應模式下 （預處理、共處理、後處理) ，以1 nM濃度TCDD混合桂枝及五味子萃取物，藉由其所產生之冷光值強度判定TCDD對AhR結合的能力，觀察TCDD與中藥萃取物之間的交互作用。
研究結果顯示，預處理具有最好之抑制效果，在四種濃度 (1、10、30、50 μg/mL) 與1 nM TCDD的預處理模式中，五味子的水及90%乙醇萃取物之平均抑制率為15及4%；桂枝的水及90%乙醇萃取物之平均抑制率為48及35%，其中50 μg/mL桂枝的水萃取物在預處理及後處理實驗中，其抑制率皆為51%為本實驗最好之抑制效果。此外，研究亦利用單一成份純物質五味子乙素及桂皮醛進行TCDD代謝反應抑制試驗，結果顯示五味子乙素及桂皮醛之平均抑制率在三種處理模式中分別達8 ~ 27%及-113 ~ 35%，初步研究顯示五味子及桂枝之水及乙醇萃取物對戴奧辛誘導肝臟代謝路徑具有部分的抑制效果。

關鍵字

人類肝癌細胞 ； 五味子 ； 桂枝 ； 戴奧辛

並列摘要

Dioxin is notorious for its liver toxicology and procarcinogenesis because of its lipophilicity and highly accumulation through food web. Most in-vitro studies are focused on the inhibition effect for food extracts and medicine on TCDD induction of CYP1A1 in rat hepatoma cell (H4IIE) or human breast cancer cell (MCF-7). Little information can be referred to the inhibition effect for Chinese herbs on TCDD induction of CYP1A1 in human hepatoam cells. The DRE-Luciferase assay was applied in this study to evaluate the inhibition effect of Schisandra chinensis and Cinnamomum cassia extracts on TCDD induction of CYP1A1 based on three treatment modes: pre-treatment, cotreatment and post-treatment.

The results showed that the highest inhibition effect for the pre-treatment among four concentrations (1, 10, 30, 50 μg/ml) was 15% for Schisandra chinensis aqueous extracts and 4% for Schisandra chinensis 90% alcoholic extracts, respectively. The highest inhibirion effect was 48% for Cinnamomum cassia aqueous extracts and 35% for Cinnamomum cassia 90% alcoholic extracts, respectively. Furthermore, Schisandrin B and Cinnamaldehyde were processed to investigate the inhibition effect on TCDD induction of CYP1A1. The results showed that the inhibition effect was among 8 ~ 27% and -113 ~ 35% for Schisandrin B and Cinnamaldehyde, respectively, for the three treatment mode.
The preliminary findings demonstrated that Schisandra chinensis and Cinnamomum cassia extracts show promising potential as an agent to block the AhR metabolic pathway for 2,3,7,8-TCDD.

並列關鍵字

Dioxin ； Cinnamomum cassia ； Schisandra chinensis ； Human hepatoma cell

參考文獻

何秀琴，2007，”桂枝湯的臨床運用”，中醫兒科醫學雜誌, 第9卷, 第1期, 46-60。

黃怡瑋、李偉山、張簡國平、林紀佑、高禎陽、李心予，2004，”戴奧辛細胞檢測系統之建立”，生物科學，第47卷，第1期， 1-18。

Androutsopoulos VP, Tsatsakis AM and Spandidos DA. 2009. Cytochrome P450 CYP1A1: wider roles in cancer progression and prevention. BioMedCentral. BMC Cancer. 9: 187.

Ayres DC and Loike JD. 1990. Lignans: chemical, biological, and clinical properties. Cambridge:Cambridge University Press. England. 12-17.

Behnisch PA, Hosoe K, Brouwer A and Sakai SI. 2002. Screening of dioxin-like toxicity equivalents for various matrices with wildtype and recombinant rat hepatoma H4IIE cells. Toxicological Sciences. 69: 125-130.