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A 90-Day Oral Toxicity Study of the Ethanol Extract from Eupatorium Japonicum Thunb and Foeniculum Vulgare in Rats

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A 90-Day Oral Toxicity Study of the Ethanol Extract from Eupatorium Japonicum Thunb and Foeniculum Vulgare in Rats Hindawi BioMed Research International Volume 2020, Article ID 6859374, 8 pages https://doi.org/10.1155/2020/6859374 Research Article A 90-Day Oral Toxicity Study of the Ethanol Extract from Eupatorium japonicum Thunb and Foeniculum vulgare in Rats Guangcheng Dai,1 Chenglu Wang,2 Wei Tang,1 Jiangyun Liu,3 and Boxin Xue 1 1Department of Urology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China 2Department of Reproductive Endocrinology, Zhejiang Provincial People’s Hospital, Hangzhou Medical College, Hangzhou 310014, China 3College of Pharmaceutical Sciences, Soochow University, Suzhou 215123, China Correspondence should be addressed to Boxin Xue; xbxurol@163.com Received 12 March 2020; Revised 7 June 2020; Accepted 19 June 2020; Published 27 June 2020 Academic Editor: Stefano Curcio Copyright © 2020 Guangcheng Dai et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Eupatorium japonicum Thunb and Foeniculum vulgare are two of the most widely used folk herbs and constituents in many traditional Chinese herbal formulas. Nonetheless, little toxicological and safety information associated with following daily repeated exposure is obtained according to previous research. The present study was performed to assess the toxicity of ethanol extract from Eupatorium japonicum Thunb and Foeniculum vulgare (EFE) in male rats administered by dietary oral gavage at target doses of 0.39, 0.78, and 1.56 g/kg body weight/day for 90 days. There were no significant adverse effects on clinical signs, body weight, food conversion efficiency, and vital hematological indices. However, some hematology and biochemical indices such as WCV, MCH, MCHC, LY, MPV, T-CHO, as well as TG revealed significant changes in Sprague–Dawley rats and organ weights in lung and spleen showed diminished in male rats. Necropsy and histopathology findings suggested that no significant differences in absolute weights were found in all organs except lung and spleen, and no treatment-related alteration was identified in any organs. All results obtained in the present study indicated that the proper use of EFE in traditional medicine at oral dosages up to 1.56 g/kg/day body weight may harbor no prolonged toxicity to rats. However, further studies of EFE are still necessary to assess its oral safety in patients. 1. Introduction species widely cultivated in almost all countries of the world (4). It is an ancient and popular herb with a long utilization Eupatorium japonicum Thunb and Foeniculum vulgare are history. Several past studies have shown that numerous infec- widely used by folk and traditional medicines in many tions disorders including bacterium, fungus, virus, and ancient Chinese herbal formulas. Eupatorium japonicum mycobacterium can be effectively controlled using Foenicu- Thunb, belonged to the family of Asteraceae in the Asterales lum vulgare (5–7). It has been suggested that Foeniculum vul- within the Angiosperms, is widespread in China, Japan, and gare has multiple biological functions such as cytoprotective, Korea. According to previous researches, the leaves and hypoglycemic, chemopreventive, hepatoprotective, and oes- stems have been shown the effect of anti-inflammatory and trogenic activities according to previous literature (8–12). vascular smooth muscle relaxant (1). Therefore, Eupatorium Recently, some researchers stated that Foeniculum vulgare japonicum Thunb are identified as the effective antivirals, can be used for management stress-related disorders because antibacterials, diuretics, painkillers, vermifuges, and carmi- of its antistress and memory-enhancing function (13). natives (2). And it is utilized for the treatment of vomiting, In our previous study, Ethanol extract from Eupatorium indigestion, nausea, and diarrhea (The Chinese People’s japonicum Thunb and Foeniculum vulgare (EFE) can regu- Pharmacopoeia, 2005) (3). Foeniculum vulgare, the oldest late the balance of sex hormone and decrease the hyperplasia valid name within the genus Foeniculum,isaflowering plant epithelia of prostate in male Sprague–Dawley (SD) rats, 2 BioMed Research International which might result in an inhibitory effect of benign prostatic during the experimental period and at the end of the study. hyperplasia (14). Nonetheless, to our knowledge, no data are Also, food consumption was calculated every four days, and available so far on the safety assessment of EFE in animals. total food consumption for each rat was assessed by summa- Therefore, the present research investigated the potential tion of the food intake recorded from every feeding period. toxicity of EFE using a 90-day feeding study in male SD rats. Furthermore, food conversion efficiency was measured as the weight gain per gram of food consumed. 2. Materials and Methods At the end of the 90-day oral feeding, rats were sacrificed after diets withdrawing 16 h. Blood from abdominal aorta 2.1. Materials were collected and divided into two tubes for blood hemato- logical and chemistry analysis, respectively. We then weighed 2.1.1. Preparation of EFE. The whole plants of Eupatorium and performed histopathological examinations on organs japonicum Thunb and Foeniculum vulgare were purchased such as heart, spleen, lung, liver, kidney, brain, thymus, from the local Chinese herb market and the voucher speci- adrenal gland, testes, and epididymis. mens were deposited in the herbarium of College of Pharma- ceutical Sciences, Soochow University (Suzhou, China). 2.4. Observations For the ethanol extract preparation, formula herbs 20 kg (Eupatorium japonicum Thunb. and Foeniculum vulgare 2.4.1. Clinical Signs. We continuously monitored the changes were marked, respectively, 1 : 1), dried and teared for prepara- every day in physical appearance, behavioral pattern, breath, tion, reflux extraction twice with 10 times 85% ethanol, 1.5 glandular secretion, excreta, mortality, and signs of illness hours every time, merge extracting solution, decompression throughout the experiment. Additionally, the body weight enrichment to a quarter of the volume and let stand for the of each rat was observed every four days in our oral toxicity night, precipitation with Buchner funnel, filtration solution study. concentration under vacuum to the density of 1.05 in the temp of 60°C, the ethanol solution is concentrated by reduc- 2.4.2. Body Weight, Food Consumption, and Food Conversion ing pressure and dried by vacuum; finally, 1.56 kg block solid Efficiency. Body weight was recorded on the first day of extract is obtained, which was utilized in the following tests. administration, then recorded every four days during the The yield of EFE was 7.8%. 90-day feeding and at the end of the experiment. Food con- sumption was evaluated over successive periods of 90 days 2.2. Experimental Animals. Forty male adult Sprague–Daw- by weighing the rats. In addition, food conversion efficiency ley rats, 6 weeks old, 124.5-142.0 g, were purchased from was measured as the weight gain per gram of food consumed. the Slac Laboratory Animal Co., Ltd. (Shanghai, China). Mice were individually kept in cages with stainless steel grid 2.4.3. Hematological and Biochemical Analyses. Hematologi- covers, bedded with sterilized wood shavings, and were cared cal parameters, including white blood cell (WBC), red blood in a controlled environment. The commercial diets (Suzhou cell (RBC), hemoglobin (Hb), five-classification of WBC Shuangshi Laboratory Animal Feed Science and Technology (Neutrophil, NE/Lymphocyte, LY/Eosinophil, EO/Monocyte, Co., Ltd. Soochow) and tap water were provided ad libitum. MO/Basophil, BA), hematocrit (HCT), mean corpuscular The rats were allowed 1 week for acclimation prior to the hemoglobin (MCH), mean corpuscular volume (MCV), mean conduction of experiments. corpuscular hemoglobin concentration (MCHC), mean plate- All animals received human care according to the Chi- let volume (MPV), and red cell distribution width-coefficient nese legislation on the usage and care of laboratory animals. of variation (RDW-CV), was analyzed by automatic hematol- The study was approved by the committee of Soochow ogy analyzers XE2100 (Sysmex, Japan). Biochemical parame- University of Science and Technology. ters such as alanine aminotransferase (ALT), total protein (TP), aspartate aminotransferase (AST), albumin (ALB), 2.3. Study Design Overview. The study was performed based blood urea nitrogen (BUN), creatinine (CERA), triglyceride on the Procedure and Methods of Food Safety Toxicological (TG), and total cholesterol (T-CHO) were measured by Cobas Assessment, GB15193.13-2003 (in Chinese) (15). 8000 Biochemistry Automatic Analyzer (Roche, Germany). 2.3.1. 90-Day Period of Feeding Study. After getting 7-day 2.4.4. Organs Weight and Histopathological Examinations. At periods of acclimatization, forty rats were used in the study. the ending of the experiment, all the rats were euthanized and The dose was discussed according to the results of previous subjected to necropsy. Pertinent observations on the pathol- and pharmacodynamic and acute oral toxicity study. All rats ogy of each organ were conducted and recorded. Organs such were randomly selected and included into three treatment as heart, lung, liver, spleen, kidney, brain, thymus, adrenal groups and a control group. The oral feeding was continually gland, testes, and epididymis were weighed before the conducted (i. g.) at the same time from Monday to
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