- 學位論文
茵陳蒿湯對大白鼠肝粒腺體脂質過氧化之抑制作用
Lipid Peroxidative Inhibition of Yin-Chen-Hao Tang on Rat Liver Mitochondria
摘要
肝病為近年來衛生署公佈之十大死亡病因之一。台灣地區肝炎感染率頗高，生活不規律、酗酒、飲食不潔、打針輸血、病毒感染、化學藥品傷害等，常易感染肝炎。由於肝病變很少像中風、心肌梗塞那樣急性發作而不省人事，許多都是經年累月肝細胞逐漸受傷害所造成之慢性疾病，而國人用中藥調治慢性疾病有愈來愈多的趨勢。因此，本研究選擇臨床常用治肝病之中藥方劑：茵陳蒿湯、小柴胡湯、大柴胡湯、八正散、梔子柏皮湯、當歸龍薈丸、黃連解毒湯、加味逍遙散、桂枝茯苓丸、龍膽瀉肝湯共十種,評估其對大鼠肝粒線體脂質過氧化物之抑制效果。 各方劑之水抽取物，經由三種不同誘導劑對大白鼠肝粒腺體所誘生脂質過氧化物，其過氧化物經反應生成MDA後，再加入TBA 作用產生MDA(TBA)2，以HPLC在532 nm波長下偵測，定量MDA(TBA)2，評估其抑制脂質過氧化之效果，結果十種肝病常用中藥處方中，對肝粒腺體脂質過氧化抑制之效果，茵陳蒿湯具有最強之抑制活性，其對t-BuOOH、Fe2+、β-NADPH所誘生脂質過氧化抑制之IC50分別為1.28、0.85、3.52 (10-2 ?g/?l)，對照組Vit.E分別為6.12、3.28、33.43 (10-2 ?g/?l)。本論文乃進一步探討茵陳蒿湯中各組成藥材：酒大黃、茵陳蒿、山梔子、酒大黃+茵陳蒿、酒大黃+山梔子、茵陳蒿+山梔子對肝粒線體脂質過氧化之抑制效果，來分析茵陳蒿湯之配佐原理，結果顯示，茵陳蒿湯中脂質過氧化抑制具最佳效果的為酒大黃，其對t-BuOOH、Fe2+、β-NADPH所誘生脂質過氧化抑制之IC50分別為1.81、0.78、3.63 (10-2 ?g/?l)，對照組Vit.E分別為9.37、6.71、27.09 (10-2 ?g/?l)。 另外，大黃在臨床調製方劑常用之飲片有生品、酒大黃、蒸大黃等，故本實驗利用上述不同炮製大黃飲片調製之茵陳蒿湯，來評估各製劑脂質過氧化抑制之效果，結果以根據傷寒論張仲景方以酒大黃配製之茵陳蒿湯效果最佳，其對t-BuOOH、Fe2+、β-NADPH所誘生大鼠肝粒腺體細胞脂質過氧化抑制之IC50分別為0.86、0.78、2.68 (10-2 ?g/?l)，對照組Vit.E分別為6.12、3.28、33.43 (10-2 ?g/?l)。由以上結果證實，根據方劑學君臣佐使之原則，證實酒大黃為茵陳蒿湯保護肝細胞抗脂質過氧化之主藥，亦證實傳統中國醫學在藥物調製過程中大黃炮製之意義。
並列摘要
Liver disease is one of the major causes of human death in Taiwan. There are several complicated factors have been demonstrated in the development of liver diseases such as virus infection, alcoholic, drug abuse, and improper perfusion. Chinese medicinal prescriptions have been used widely in treatment of liver diseases for thousands of years and showed the effective therapeutic effect. However, the scientific evidences for Chinese medicinal prescriptions on therapy of liver diseases are still undefined. In the present study, ten popular Chinese prescriptions including yin-chen-hao tang, xiao chai-hu tang, da chai-hu tang, ba zheng san, zhi-zi bo-pi tang, dang-gui long lui wan, huang-lian jie du tang, jia wei xiao yao san, gui-zhi fu-ling wan and long-dan xie gan tang were used to investigate their antioxidative activities in rat liver mitochondria. To evaluate the antioxidative activities of prescriptions as described above, in vitro lipid peroxide formation induced by several stimulators in rat liver mitochondria was performed and the level of lipid peroxidation was measured by the amount of MDA(TBA)2 complex formation detected by HPLC at wavelength 532 nm. The results revealed that yin-chen-hao tang showed the significant inhibitory effect on t-BOOH, Fe2+, β-NADPH-induced lipid peroxidation, and IC50 values on each stimulator are 1.28, 0.85 and 3.52 (10-2 ?g/?l), respectively. There are three components including alcoholic da-huang (Rhei Rhizoma), yin chen hao (Artemisiae Capillaris Herba), and zhi zi (Gardeniae Fructus) were involved in the prescriptions of yin-chen-hao tang. Therefore, it is interesting to find out which component is major for the antioxidative activity in yin-chen hao tang. As the same part of experiment, the results appeared that alcohoic da-huang is the most potent among these three components, and show the dose-dependent inhibition on t-BOOH, Fe2+, and ?-NADPH-induced lipid peroxidation. The IC50 values of alcoholic da huang on inhibition of each stimulator induced lipid peroxidation are 1.81, 0.78 and 3.63 (10-2 ?g/?l), respectively. In Chinese medicine, da-huang was cured commonly by different processings such as unhandled, alcoholic, and steamed one. Therefore, evaluation of the anti-lipid peroxidative activities of yin-chen-hao tang with different kinds of da-huang was performed in the following study. The data appeared that alcoholic da-huang used in yin-chen-hao tang showed the most potent anti-lipid peroxidative activities on t-BOOH, Fe2+, and ?-NADPH induced lipid peroxidation, and IC50 values of each stimulator were 0.86, 0.78, 2.68 (10-2 ?g/?l), respectively. In ancient book “shang han lun”, yin-chen hao tang had been described to exert the potent therapeutic effect on liver diseases. In this study, we provided firstly the scientific evidence to demonstrate the antioxidative activity of yin-chen hao tang in rat liver mitochondria. Based on these data, we proposed that yin-chen hao tang is a Chinese prescription with significant antioxidative activities, and may used as a protectant for liver disease. It should be deserved for further study and for clinical trial in the future.