三黃瀉心湯及其相關組成之抗癌和抗病毒活性研究

Study of anti-tumor and anti-virus activities of San-Ou-Sya-Sin-To and its related ingredients

劉純純(Chun-Chun Liu)

指導教授 ： 蔣連財 ； 林俊清

高雄醫學大學/醫學院/醫學研究所/碩士(2005年)

摘要

三黃瀉心湯是常用傳統中藥方劑，常用於治療血壓高、便秘、及胃潰瘍等疾病。其單方都具有抗發炎、抗癌、抗病毒等作用，因此本研究實驗目的係利用XTT還原法研究三黃瀉心湯及其相關成分抗癌（血癌及固形癌）及抗病毒（呼吸道融合病毒、克沙奇病毒 B1和疱疹病毒第一型）活性分析。
抗癌實驗結果顯示，複方劑兩種萃取法對於毒殺兩株血癌細胞的活性並沒有明顯得差異，而黃連及大黃甲醇粗萃物抑制K562的活性比水抽粗萃物好。複方和單方抑制固形癌細胞株試驗結果：抑制腸癌細胞株（HT-29）的活性都優於抑制肺癌細胞株（A549），雖然如此，但是其抗癌的活性並不顯著。
抗病毒結果顯示，三黃瀉心湯甲醇粗萃物對於呼吸道融合病毒、克沙奇病毒B1以及疱疹病毒第一型都具有抑制活性，抑制百分之五十 (EC50) 所需濃度分別為74.3±0.1、4.6±0.3及35.6±0.7μg/ml；在單方中，大黃甲醇粗萃物對呼吸道融合病毒有良好的抑制活性，其EC50值為9.8±1.1μg/ml；而黃連及黃芩甲醇粗萃物具有抑制克沙奇病毒B1的活性，其EC50值分別為6.9±0.1及11.6±0.2μg/ml；另外，大黃兩種粗萃物均有抑制疱疹病毒第一型感染活性，水抽和甲醇抽粗萃物之EC50值分別為35.5±1.3及20.5±0.7μg/ml。
抑制呼吸道融合病毒（RSV）感染的時間效應研究結果，顯示三黃瀉心湯具有治療和預防RSV感染的活性。黃芩水粗萃物對RSV具有預防病毒感染的活性；大黃水抽及甲醇抽粗萃物，則有預防及治療RSV病毒感染的活性；另外，三黃瀉心湯及大黃能抑制呼吸道融合病毒的融合現象、吸附和穿透作用，證實具有預防病毒的感染活性和能夠抑制呼吸道融合病毒感染後的複製。大黃的純品中chrysophanol、emodin和rhein具有預防呼吸道融合病毒感染的活性，而sennoside A則是抑制呼吸道融合病毒感染後之增殖。
綜合以上結果，三黃瀉心湯及其相關成分的水抽及甲醇抽粗萃物，以及大黃已知相關成分（chrysophanol、emodin、sennoside A及rhein）都具有抗病毒的活性，顯示此複方、單方和其純品生藥應用於病毒的預防和治療上具有無窮的潛力，值得進一步深入研究，開發新的抗病毒藥物。

關鍵字

血癌 ； 固形癌 ； 三黃瀉心湯 ； 克沙奇病毒 B1 ； 呼吸道融合病毒 ； ?疹病毒第一型

並列摘要

Abstract
San-Ou-Sya-Sin-To(SOSST,San-Huang-Shel-Shin-Tang,San-Huang-Xie-Xin-Tang, 三黃瀉心湯), a traditional Chinese medicine, has been used for treatment a variety of ailments including hypertension, constipation and gastric ulcer. The ingredient crude drugs have been reported to possess anti-inflammatory, anti-cancer or anti-viral activities. The aim of this study was to investigate the in vitro anti-cancer（leukemia and solid cancer cells）and anti-virus（respiratory syncytial virus,RSV; coxsakie virus B1 , CVB1 and herpes simplex virus type-1, HSV-1）activities by XTT reduction method.
The results of the cytotoxic activities by two crude extracts from SOSST showed that were not different between two leukemia cell lines. In anti-k562 leukemia cells, the methanolic extracts of Coptis chinensis and Rheum officinale were more active than aqueous extracts. The cytotoxic results against human solid cancer cell lines by SOSST and related crude drugs exhibited that colon adenocarcinoma (HT-29) cells was more sensitive than lung carcinoma (A549) cells.
The results of the antiviral activities, the methanolic extract from SOSST possessed to inhibit infection of RSV, CVB1 and HSV-1, the 50% effective concentration (EC50) were 74.3±0.1, 4.6±0.3 and 35.6±0.7μg/ml, respectively. In related crude drugs, the methanolic extract from R. officinale expressed potent activity against RSV infection with the EC50 value of 9.8±1.1μg/ml. The antiviral activity (EC50) against CVB1 infection of the methanolic extracts from C. chinensis and S. baicalensis were 6.9±0.1及11.6±0.2μg/ml, respectively. The inhibition (EC50) of HSV-1 infection by R. officinale showed that aqueous extract was 35.3±1.3μg/ml and methanolic extract was 20.5±0.7μg/ml.
The results of the time-of-addition studies showed that SOSST could prevent and inhibit RSV infection. The aqueous extract from S. baicalensis prevented the infection of RSV, whereas the aqueous and methanolic extracts from R. officinale could prevent RSV infection and inhibit the replication after infection. The methanolic extracts from SOSST and R. officinale could inhibit the viral attachment, penetration, and syncytial formation induced by RSV, which confirmed the prophylactic activity against RSV infection and inhibited the virus replication after infection. The R. officinale-related pure compounds, chrysophanol, emodin and rhein, could prevent RSV virus infection, whereas sennoside A inhibited virus replication after RSV infection.
In conclusion, this study showed that the aqueous and methanolic extracts from SOSST and its ingredients and R. officinale-related pure compounds (chrysophanol, emodin, sennoside A and rhein) possessed anti-virus activities, suggesting the potential use of these drugs for prevention and treatment of viral infections. Further investigation to develop new antiviral drugs from these Chinese medicines and its pure compounds are warranted.

並列關鍵字

coxsakie virus B1 ； solid cancer cells ； herpes simplex virus type-1 ； San-Ou-Sya-Sin-To ； leukemia cancer ； respiratory syncytial virus

參考文獻

文獻

Abe, I., Seki, T., Noguchi, H., and Kashiwada, Y.: Galloyl esters from rhubarb are potent inhibitors of squalene epoxidase, a key enzyme in cholesterol biosynthesis. Planta Med 66: 753-6, 2000

Agarwal, S. K., Singh, S. S., Verma, S., and Kumar, S.: Antifungal activity of anthraquinone derivatives from Rheum emodi. J Ethnopharmacol 72: 43-6, 2000

Alcaraz, M. J. and Ferrandiz, M. L.: Modification of arachidonic metabolism by flavonoids. J Ethnopharmacol 21: 209-29, 1987

Alrabiah, F. A. and Sacks, S. L.: New antiherpesvirus agents. Their targets and therapeutic potential. Drugs 52: 17-32, 1996