Sprague dawley rats

Adult male SD (Sprague Dawley) rats were purchased from Central Lab Animal Inc. (Seoul, Korea). Adult male and female ICR (Institute of Cancer Research) mice were purchased from Orient Inc. (Gyeonggi-do, Korea). Mouse embryos were isolated at E12.5 and E13.5. All animal procedures were carried out in accordance with the Guide for the Care and Use of Laboratory Animals of the Ministry of Agriculture, Food, and Rural Affairs (MAFRA), and were approved by the Institutional Animal Care and Use Committee (IACUC) at Yonsei University (license #2017-0230, Date of approval March 10, 2020; and #2015-0147, Date of approval July 1, 2016). All animal procedures were performed under veterinary supervision according to the guidelines stipulated by the Ethical Committee. Mice and rats used in these studies were cared for in accordance with the National Institutes of Health “Guide for the Care and Use of Laboratory Animals” and ARRIVE guidelines.

Pregnant Sprague–Dawley rats (gestation day 14) were purchased from Central Lab. Animal Inc. (Seoul, Korea). The rats were individually housed with a 12 h light/dark cycle and a standard temperature (22 ± 2 °C) with food and water freely available. Neonatal maternal separation was carried out based on previous studies [70 (link),71 (link)]. The day of delivery was designated as PND 0. After birth, litters were randomly allocated to the control or maternal separation (MS) groups. A total of 46 pups were assigned to four groups: Con-males (n = 12), MS-males (n = 12), Con-females (n = 12) and MS-females (n = 10). During PNDs 1–21, the MS pups were separated twice a day, for 180 min each time (for a total of 360 min per day), from their dam nest (09:00–12:00 a.m. and 1:00–4:00 p.m.) and placed into another clean cage with a heating pad (30 ± 2 °C). After each 180 min, the pups were returned to their dams until the next period of separation. The control pups remained in their home cage with their dams. On PND 22, all the pups were weaned and separated by sex (2–3 rats per cage) until PND 56. All the animal experiments were conducted in accordance with the animal care guidelines of the National Institute for Health (NIH) Guide, and approved by the Animal Care and Use Committee at Kyung Hee University (KHUASP-17-156).

One hundred male Sprague–Dawley (SD) rats (250–280 g) were purchased from Central Lab Animal Inc. (Seoul, Korea) randomly assigned into groups. The experimental animals were given free access to standard diet and water in rooms maintained at 25 °C on 12-h light/dark cycles. Four rats were sacrificed for preliminary CCl₄ toxicity testing, and 12 of the surviving rats were placed in each of the following groups. Control rats were injected with olive oil (vehicle; 0.1 ml/kg body weight, i.p.) for 72 h. The curcumin group was intragastrically administered with 200 mg/kg curcumin for 72 h. The CLL group was intragastrically administered with 300 mg/kg CLL extract. The CCl₄ group received CCl₄ (0.1 ml/kg, i.p.) for 72 h. The CCl₄-curcumin group received curcumin (200 mg/kg) intragastrically before 3 days of CCl₄ treatment. The CCl₄-CLL groups received 100 mg/kg, 200 mg/kg, or 300 mg/kg of CLL extract intragastrically before 3 days of CCl₄ treatment. All of the experimental protocols conducted on rats were performed in accordance with internationally accepted principles for laboratory animal use and care and approval was obtained from the Care and Use of Laboratory Animals Committee of Chonbuk National University Hospital. All procedures were also approved by the Institutional Animal Care and Use Committee of Chonbuk National University Hospital (IACUC protocol CBU 150608-25).