Mol Cell Toxicol (2010) 6:239-246 DOI 10.1007/s13273-010-0033-2 Abstract The short-term toxicity of PAI-N002, a mixture of Schizandra chinensis, Astragalus mem- branaceus, Artemisia capillaris, and Coriolus versi- color extracts (1 : 1 : 1 : 1), was evaluated in rats. This study also investigated the protective effect of PAI- N002 on liver injury induced by the co-administration of ethanol and carbon tetrachloride (EC) in rats. PAI- N002 was virtually non-toxic to rats after a single oral administration, with LD 50 values ¤5,000 mg/kg. The subacute toxicity study showed that 2-week repeated oral dose of PAI-N002 did not cause any adverse ef- fects on clinical signs, body weight, food consump- tion, ophthalmoscopy, hematology, serum biochem- istry, gross findings, and organ weights in rats given up to 1,000 mg/kg/day. When rats with EC-induced hepatotoxicity were treated with PAI-N002 at 250 mg /kg/day for 28 days, PAI-N002 treatment significantly improved EC-induced hepatic injury, as evidenced by the decline of serum AST and ALT activities and de- creased histopathological alterations. PAI-N002 also had an antioxidant benefit by decreasing the lipid per- oxidative product malondialdehyde and increasing the levels of the antioxidant glutathione and the activities of the antioxidant enzymes catalase, superoxide dis- mutase, and glutathione-S-transferase. These results indicate that the short-term treatment of rats with PAI- N002 has no harmful effects and that PAI-N002 has hepatoprotective and antioxidant properties in EC- induced chronic liver injury in rats. Keywords PAI-N002, Ethanol, Carbon tetrachloride, Hepatotoxicity, Protective effect Herbal products have gained in popularity over the last decade, and are now used by approximately 20% of the population in the United States 1 . In developing countries, as much as 80% of the indigenous popula- tions depend on medicinal plants and traditional sys- tems of medicine as their primary source of health- care 2 . It has been reported that a number of herbal pro- ducts protect against liver injury, and many possess one or a combination of antioxidant, antifibrotic, im- mune modulatory, or antiviral properties 3,4 . In recent years, there has been a substantial increase in the use complementary and alternative therapies that utilize herbal medicines by patients with liver disease 5-7 . PAI-N002 is a mixture of the Schizandra chinensis, Astragalus membranaceus, Artemisia capillaris, and Coriolus versicolor extracts (1 : 1 : 1 : 1) developed by PHARMAPEX Co. (Suwon, Korea). Schizandra chi- nensis has many purported medicinal properties 8 , but the most substantial body of research focuses on its effect on hepatic dysfunction 9-11 . Astragalus membra- naceus is used commonly as a diuretic or a tonic in Oriental medicine. It is also used for the treatment of oxidative stress, liver fibrosis, immune diseases, and cardiovascular diseases 12-14 . Artemisia capillaris is commonly used to treat diseases of the liver and gall- bladder in traditional oriental medicine 15,16 . Coriolus versicolor extract is used to treat liver cancer and pul- monary infections, and to support liver health 17,18 . The pharmacological effects of these herbal products have been extensively studied, but there is limited sci- entific evidence to establish the safety and efficacy of ORIGINAL PAPER Evaluation of the toxicological properties and hepatoprotective effects of PAI-N002, a mixture of herbal extracts, in rats Sung-Hwan Kim 1 , Jeong-Hyeon Lim 1 , In-Sik Shin 1 , Changjong Moon 1 , Soo-Hyun Park 1 , Sung-Ho Kim 1 , Jung-Sik Lee 2 , Eun-Hye Kwon 2 & Jong-Choon Kim 1 Received: 30 March 2010 / Accepted: 21 June 2010 �The Korean Society of Toxicogenomics and Toxicoporeteomics and Springer 2010 1 College of Veterinary Medicine, Chonnam National University, Gwangju 500-757, Korea 2 PHARMAPEX Co., Suwon, Gyeonggi 443-270, Korea Correspondence and requests for materials should be addressed to J. C. Kim ( toxkim@jnu.ac.kr)