黃連解毒湯及所含生藥組成抑制B型肝炎病毒蛋白及抗氧化活性研究

Inhibition of HBeAg Expression and Antioxidant Activity of Huanglian-jie-du-tang and its ingredients

華宜蓁(Yi-Cheng Hua)

指導教授 ： 林俊清

高雄醫學大學/藥學院/天然藥物研究所碩士班/碩士(2005年)

摘要

為了探討黃連解毒湯抗B型肝炎病毒蛋白以及抗氧化活性，本研究以B型肝炎病毒穩定轉植肝癌細胞株HepG2 2.2.15來測試黃連解毒湯對B型肝炎病毒蛋白的抑制活性。經由ELISA分析來定量細胞外懸浮液e抗原(HBeAg)及表面抗原(HBsAg)量，以XTT方法測試藥物對細胞的毒殺作用；自由基的清除及抑制分別由Cytochrome C還原比色法及xanthine oxidase 抑制試驗來測試。根據本實驗結果，黃連解毒湯對於HepG2 2.2.15分泌HBeAg至細胞外懸浮液有良好的抑制活性，其抑制百分之五十(IC50)的HBeAg 所需濃度為 82.87μg/ml，同時顯示黃連解毒湯在抑制HBeAg所需濃度下對HepG2 2.2.15並不具有細胞毒殺及增生抑制的活性。毒殺百分之五十細胞(CC50)濃度為 299.67μg/ml，其選擇指數 (selectivity index, SI )為1.92；在此結果發現梔子亦有抗HBeAg活性，而黃芩、黃連、黃柏對e抗原或表面抗原都無抑制作用。另外，黃連解毒湯在自由基清除能力及自由基抑制能力方面其IC50分別為6.24及5.51 mg/ml；在此結果發現梔子亦有清除及抑制自由基能力，而黃芩、黃連、黃柏在自由基清除及自由基抑制能力上皆較黃連解毒湯及栀子弱。因此著者推論黃連解毒湯抗HBeAg及抗氧化活性係由梔子此單位生藥為主軸，而其抗B型肝炎病毒蛋白與抗氧化活性可能有關。

關鍵字

e抗原 ； 抗氧化 ； 黃蓮解毒湯

並列摘要

To examine the effects on the expression of hepatitis B virus protein and antioxidant activity of Huanglian-jie-du-tang (HLJDT) and its ingredients, HepG2 2.2.15 was used to detect the amount of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) by Enzyme-linked immunosorbent assay. The cytotoxicity of studied extracts was evaluated by XTT assay. The free radical scavenging activity and anti-superoxide radical formation was assayed by the cytochrome C reduction method and xanthine/xanthine oxidase system, respectively. The results showed that HLJDT inhibited the production of HBeAg (p<0.05) with an IC50 of 82.87 μg/ml. However, HLJDT did not have any effect on HBsAg. The 50% cytotoxic concentration (CC50) of HLJDT against HepG2 2.2.15 cells was 299.67 μg/ml with a selectivity index of 1.92. HLJDT and its ingredients all possessed anti-oxidant activity. The IC50 of free radical scavenging activity and anti-superoxide formation of HLJDT was 6.24 and 5.51 mg/ml, respectively. HLJDT possessed both antioxidant and anti-HBeAg activities and might be beneficial against chronic HBV infection. Hence, it supported activity that inhibition of HBeAg expression because of Gardenia jasminoides Ellis.

並列關鍵字

HBeAg ； Antioxidant ； HLJDT

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