Myelophil attenuates brain oxidative damage by modulating the hypothalamus-pituitary-adrenal (HPA) axis in a chronic cold-stress mouse model Hyeong-Geug Kim a , Jin-Seok Lee a , Jong-Min Han a , Jong-Seok Lee b , Min-Kyung Choi a , Seung-Wan Son c , Yong-Kyung Kim d , Chang-Gue Son a,n Q1 a Liver and Immunology Research Center, Daejeon Oriental Hospital of Oriental Medical College of Daejeon University, 22-5 Daehung-dong, Jung-gu, Daejeon 301-724, Republic of Korea b Gyeonggi Biocenter, Gyeonggi Institute of Science & Technology Promotion (GSTEP), Suwon, Gyeonggi-do 443-270, Republic of Korea c Department of Biomedical Engineering, College of Health Science, Korea University, Seongbuk-Gu, Seoul 136-703, Republic of Korea d Laboratory of Endocrine Cell Biology, Department of Internal Medicine, Chungnam National University School of Medicine, 640 Daesa-dong, Daejeon 301-721, Republic of Korea article info Article history: Received 27 December 2012 Received in revised form 12 April 2013 Accepted 23 April 2013 Keywords: Oxidative stress Brain damage Myelophil Chronic cold stress abstract Ethnopharmacological relevance: Myelophil is composed of Astragali Radix and Salviae Miltiorrhizae Radix, according to the long traditional pharmacological practices, and it has been used for patients with chronic fatigue-associated symptoms including concentration problem or memory loss. Aim of the study: This study aimed to evaluate the clinical relevance of Myelophil on brain oxidative damage using a chronic cold stress mice model. Material and methods: Balb/c mice were subjected to cold stress (4 1C for 4 h) six times per week for 2 weeks with or without oral administration of Myelophil (50, 100, or 200 mg/kg), or ascorbic acid (50 mg/kg). Results: Chronic cold stress induced histopathological hippocampal apoptosis with drastically increased serum levels of total reactive oxygen species and nitric oxide, as well as brain lipid peroxidation levels, protein carbonyl, and caspase-3/7 activity. These alterations were significantly ameliorated by Myelophil treatment. Myelophil administration significantly recovered the depleted glutathione and its enzymes, superoxide dismutase activity, and catalase protein and gene expression levels. Serum levels of corticosterone, dopamine, and adrenaline were notably altered by chronic cold stress but were significantly ameliorated by Myelophil treatment. Myelophil also normalized alterations in tumor necrosis factor-α, interleukin (IL)-1β, and IL-10 gene expression and protein levels. Chronic cold stress up-regulated gene expression levels of phenylethanolamine N-methyltransferase and monoamine oxidase-B, and glucocorticoid receptors in the hypothalamus and hippocampus, respectively, whereas Myelophil treatment completely normalized these levels. Conclusions: These results suggest that Myelophil has potent pharmaceutical effects against chronic cold-stress-induced brain damage by relieving oxidative stress and inflammation and regulating stress hormones in mice. & 2013 Published by Elsevier Ireland Ltd. 1. Introduction Myelophil, an extract mixture of Astragali Radix and Salviae Miltiorrhizae Radix, was developed according to the long tradi- tional pharmacological practices, which has been used for patients with chronic fatigue-associated symptoms including concentra- tion problem or memory loss (Cho et al., 2009). Astragali Radix and Salviae Miltiorrhizae Radix are the representative herbs with func- tions of maintaining the homeostasis of qi and blood, respectively. Qi and blood are two essential components of the human body as defined by the traditional theory of the Traditional Chinese Medicine (TCM) (Takeichi et al., 1999). In the TCM, Astragali Radix and Salviae Miltiorrhizae Radix have been used for enhancement of brain functions and brain-associated disorders such as cerebrovascular disease, degenerative disorder and chronic fatigue syndrome (Zhang and Huang, 1990). Experimental data also support that Astragali Radix and Salviae Miltiorrhizae Radix, the two components of Myelophil, have anti-stress and anti-amnesic effects in brain (Lee et al., 2012). On the other hand, stress is a common and unavoidable phenomenon in human life, and excess stress influences physio- logical homeostasis affecting mental and physical health. Stress is 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 Contents lists available at SciVerse ScienceDirect journal homepage: www.elsevier.com/locate/jep Journal of Ethnopharmacology 0378-8741/$ - see front matter & 2013 Published by Elsevier Ireland Ltd. http://dx.doi.org/10.1016/j.jep.2013.04.046 n Corresponding author. Tel.: +82 42 229 6723; fax: +82 42 254 3403. E-mail address: ckson@dju.ac.kr (C.-G. Son). Please cite this article as: Kim, H.-G., et al., Myelophil attenuates brain oxidative damage by modulating the hypothalamus-pituitary- adrenal (HPA) axis in a chronic.... Journal of Ethnopharmacology (2013), http://dx.doi.org/10.1016/j.jep.2013.04.046i Journal of Ethnopharmacology ∎ (∎∎∎∎) ∎∎∎–∎∎∎