中醫傳統草藥方劑-龍膽瀉肝湯變方用於大鼠急性肝損傷模式之解毒與抗氧化療效評估

The Evaluation of Antioxidation and Detoxification of Traditional Chinese Herbal Drug – Modified Long-Dan-Xie-Gan-Tang in Acute Hepatic Injury Rat Model

陳俊升(Jiun-Sheng Chen)

指導教授 ： 曾惠芬

共同指導教授 ： 邱雲棕(Yung-Tsung Chiu)

國立暨南國際大學/科技學院/生物醫學科技研究所/碩士(2007年)

https://doi.org/10.6837/NCNU.2007.00050

摘要

龍膽瀉肝湯為中國傳統方劑，廣泛的應用在急性肝炎、膽道炎及膀胱炎等疾病上，本研究利用大鼠急性肝損傷模式來評估龍膽瀉肝湯變方應用於保肝及抗氧化效果，大鼠共分為六組分別為正常對照組、四氯化碳組、低劑量中藥控制組、高劑量中藥控制組、低劑量中藥四氯化碳組及高劑量中藥四氯化碳組（111 and 333 mg/kg）。餵食中藥組預先餵食低、高劑量龍膽瀉肝湯變方七天，第八天再依照各組設計，以50 %四氯化碳（2 ml/kg）誘導肝損傷。利用血清生化值檢查（AST、ALT）、組織切片檢查、西方墨點法分析細胞色素P450相關亞型（2E1、3A1及3A2）表現量、抗氧化酵素（superoxide dismutase, SOD、catalase, CAT、glutathione peroxidase, GPx）以及麩胱甘肽硫轉移酶（glutathione S-transferase）活性測量等，來探討龍膽瀉肝湯變方之藥理機制；再以clone 9細胞株試驗來驗證動物模式之結果。實驗結果發現在高劑量中藥處理組中ALT酵素量顯著高於四氯化碳組（716.6 ± 704.0, 216.6 ± 138.2 IU/l）（P<0.05），組織學中亦發現中藥處理組之空泡樣變性程度較四氯化碳組嚴重，表示龍膽瀉肝湯變方會增強四氯化碳之肝毒性。此外，與正常對照組比較下，中藥控制組細胞色素CYP 2E1、3A1與3A2亞型之表現量顯著上升，抗氧化酵素SOD及GPx活性下降。在中藥處理組亦可觀察到Phase I之CYP 3A1與3A2酵素表現量較四氯化碳組高，其他抗氧化酵素和Phase II酵素GST則無顯著影響。在細胞實驗中龍膽瀉肝湯變方刺激CYP 2E1表現量增加，且無法有效降低氧化壓力；與動物實驗結果相符。故推測服用龍膽瀉肝湯變方後，由於Phase I酵素P450表現量增加、且減低GPx、SOD活性，導致四氯化碳造成之肝毒性加劇。

關鍵字

龍膽瀉肝湯 ； 急性肝損傷動物模式 ； 細胞色素P450 ； 抗氧化

並列摘要

Long-Dan-Xie-Gan-Tang (LDXGT), a traditional herbal drug, is widely used in various diseases, such as acute hepatitis,cholangitis and cystitis. We investigated the detoxification and antioxidative effects of modified LDXGT in rat acute liver injury model. Rats were grouped into six groups: Control, CCl4, drug-Ctrl (111 and 333 mg/kg) and drug-CCl4 groups. Groups which pretreated with LDXGT for 7 consecutive days were administered either with or without 50 % CCl4 (2 ml/kg) to induce liver injury. The detoxification and antioxidative effects were evaluated by serological tests (AST and ALT), pathological observation, the content of cytochrome P450 (CYP) subfamilies (2E1, 3A1 and 3A2), and the activities of anti-oxidative enzymes (superoxide dismutase; SOD, catalase; CAT, glutathione peroxidase; GPx) and glutathione S-transferase (GST). Moreover, clone 9 cells were used to confirm the pharmacologic effect of LDXGT. The ALT levels of drug-CCl4 groups were significantly higher than those of CCl4 group (716.6 ± 704.0, 216.6 ± 138.2 IU/l). In pathological observation, the level of ballooning degeneration in drug-CCl4 group was higher than that in CCl4 group. It indicates that LDXGT increases the toxicity of CCl4 in the rat model. Besides, the content of CYP subfamilies (CYP 2E1, CYP 3A1, CYP 3A2) were elevated while the activities of GPx and SOD were markedly decreased in drug-Ctrl groups compared with Control group. The drug-CCl4 groups also showed higher CYP subfamilies expression than CCl4 group; however, the enzymes of oxidative defense system and GST were not affected. Our data from in vitro experiments showed that LDXGT was able to induced CYP2E1 overexpression in clone 9 cells but not effective against oxidative stress, which is identical with that from animal studies. It suggests that LDXGT enhances the toxicity of CCl4 perhaps by increasing the content of CYP subfamilies and decreasing the activities of GPx and SOD.

並列關鍵字

Long-Dan-Xie-Gan-Tang ； rat acute liver injury model ； cytochrome P450 ； antioxidation

參考文獻

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