中草藥一般毒性評估模式探討

STUDIES ON COMMON TOXICITY OF CHINESE HERBAL MEDICINES

本研究以四種等倍劑量常用八種藥材（黑順片、薑半夏、法半夏、川大黃、酒大黃、青蒿、旋覆花、射干）水萃取物口服給予小鼠，觀察小鼠之死亡隻數及死亡情形，再以Litchfield& Wilcoxon方法求其50﹪致死劑量；其次再以臨床一般用量及五倍用量每天給予小鼠中藥材萃取物一次，連續給予28天，觀察其28天之亞急性毒性，以探討常用六味藥材是否具有口服急性毒性及口服給藥28天之亞急性毒性。並進行對沙門氏菌（Salmonella typhimurium）TA98、TA100及TA1535菌株致突變性之Ames試驗及對HepG2、MDCK、NRK-52E三種肝、腎細胞株的細胞毒性試驗（MTT test）。結果顯示，採樣自台灣北中南三家大盤中藥代理商之常用八種藥材中黑順片之小鼠口服LD50劑量分別為10.6 g/kg、11.9 g/kg及9.8 g/kg，其餘藥材之小鼠口服LD50劑量均大於10.0 g/kg仍未見死亡。並將此八種藥材以一般常用劑量口服給與小鼠，每天給藥一次，連續給28天後並未見有小鼠死亡、sGOT、sGPT、BUN、creatinine值及體重顯著變化；給予五倍常用劑量射干及法半夏小鼠之sGOT增加，給予五倍常用劑量射干小鼠之sGPT增加，肝組織中央靜脈區有充血，細胞脹大及變性現象，給予五倍常用劑量薑半夏及法半夏小鼠之BUN值增加，給予五倍常用劑量青蒿及法半夏小鼠之creatinine值增加，腎組織有充血，細胞腫脹變性及髓質出血現象。大黃、旋覆花、青蒿的水萃物在最高濃度（1 mg/ml）對HepG2造成細胞毒性，旋覆花在最高濃度（1 mg/ml）對MDCK造成細胞毒性，青蒿、大黃、射干、旋覆花在最高濃度（1 mg/ml）對NRK-52E造成細胞毒性。綜合以上結果顯示薑半夏、法半夏、川大黃、酒大黃、青蒿、旋覆花、射干等之小鼠口服急性毒性甚低，此八種藥材在一般劑量下並未見有亞急性毒性。附子、半夏、大黃、青蒿、旋覆花、射干六味藥材之水萃取物對沙門菌回復突變之Ames試驗並不具有致突變性，但大黃、青蒿、旋覆花、射干會造成HepG2，MDCK，NRK-52E細胞毒性，可能與細胞株的敏感性有關。The aims of this study were to investigate the oral 50% lethal toxicity and liver and kidney subacute toxicity for 28 administration of six species of common used Chinese Medicines [Aconiti Radix (AR), Rhei Rhizoma (RR), Pinelliae Rhizoma (PR), Artemisiae Herba (AH), Inulae Flos (IF), Belamcandae Rhizoma (BR)] purchased from three dealer in TCM in mice. We also investigated the anti-mutagenic effect of eight species of common used Chinese Medicines by using the NQNO and B[a]P mutagens to induce mutation in TA98, TA102, TA1535 cell lines (Ames test) and MTT test in HepG2, MDCK and NRK-52E cell lines. The results shown that the oral LD50 of the eight species of common used Chinese Medicines purchased from three dealers in TCM are very low (>10.0 g/kg), except Aconiti radix which the oral LD50 was 10.6 g/kg、11.9 g/kg and 9.8 g/kg, respectively. The 28 days feeding toxicity of the eight species of common used Chinese Medicines under the common dosage was very low in mice. However, BR increased the activities of sGOT and sGPT after five times dosage administration for 28 days in mice. Fa-PR increased the sGOT activity after five times dosage administration for 28 days in mice. The morphology of liver tissue produced congestion in central veins, cell enlarge and degeneration. The Fa-PR and Jung-PR increased the BUN level after five times dosage administration for 28 days in mice. The Fa-PR and AH increased the creatinine level after five times dosage administration for 28 days in mice. The morphology of kidney tissue produced congestion, medulla bleeding and degeneration. In the Ames test, the eight species of common used Chinese Medicines did not induce mutation in the TA98, TA100 and TA1535 cell line. RR, AH, IF (1 mg/ml) could induce cell toxicity in HepG2, AH (1 mg/ml) could induce cell toxicity in MDCK, RR, AH, IF, BR (1 mg/ml) could induce cell toxicity in NRK-52E. These results suggested that the eight species of common used Chinese Medicines possessed low acute toxicity and sub-acute toxicity under clinical dose. The AR, PR, RR, AH, IF and BR possessed non genetic mutation in bacteria. The RR, AH, IF and BR possessed HepG2, MDCK and NRK-52E toxicities. The different results in animal and cell culture may be related to the sensitivity of cell lines which were selected in this study.