Protective role of Cytoglobin and Neuroglobin against the Lipopolysaccharide (LPS)-induced inflammation in Leydig cells ex vivo |
| |
| Affiliation: | 1. Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Istanbul, Turkey;2. Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute (NHLBI)/National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD, USA;1. College of Animal Science and Technology, Beijing University of Agriculture, Beijing, 102206, China;2. College of Economics and Management, Beijing University of Agriculture, Beijing, 102206, China;3. China National Biotec Group Company Limited, Beijing, 102206, China;1. Ojinmed IVF Center, Ulaanbaatar, Mongolia;2. Department of Molecular Biology and Genetics, School of Biomedicine, Mongolian National University of Medical Sciences, Ulaanbaatar, Mongolia;3. Department of Biochemistry, School of Medicine, International University of Health and Welfare, Narita, Japan;1. Department of Science and Education, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;2. The Reproductive Medical Center, The Second Nanning People’s Hospital, Nanning, 530031, Guangxi, PR China;3. Department of Reproductive Medicine, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;4. Department of Clinical Laboratory, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;5. Guangxi University of Chinese Medicine, Nanning, 530200, Guangxi, PR China;1. Department of Gynecology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, China;2. Department of Dermatology, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, China;3. Department of Medical Imaging, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, China;1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, 730070, Gansu, China;2. Gansu Key Laboratory of Animal Generational Physiology and Reproductive Reg-ulation, China;1. Department of Reproductive Medicine Center, Zibo Maternal and Child Health Hospital, Zibo, China;2. Department of Newborn Screening Center, Zibo Maternal and Child Health Hospital, Zibo, China |
| |
| Abstract: | Leydig cells are responsible for testosterone production in male testis upon stimulation by luteinizing hormone. Inflammation and oxidative stress related Leydig cell dysfunction is one of the major causes of male infertility. Cytoglobin (CYGB) and Neuroglobin (NGB) are two globin family member proteins which protect cells against oxidative stress.In the current study, we established a Lipopolysaccharide (LPS)-induced inflammation model in TM3 Leydig cell culture to study the function of CYGB and NGB proteins under inflammatory conditions. CYGB and NGB were downregulated using siRNA and shRNA based experimental strategies. Overexpression was conducted using lentiviral pLenti-III-CYGB-2A-GFP, and pLenti-III-NGB-2A-GFP vector systems. As testicular macrophages regulate immune function upon inflammation and steroidogenesis of Leydig cells, we generated direct/indirect co-culture systems of TM3 and mouse macrophage (RAW264.7) cells ex vivo.Downregulation of CYGB and NGB induced nitride oxide (NO) release, blocked cell cycle progression, reduced testosterone production and increased inflammatory and apoptotic pathway gene expression in the presence and absence of LPS. On the other hand, CYGB and NGB overexpression reduced TNFα and COX-2 protein expressions and increased the expression of testosterone biogenesis pathway genes upon LPS stimulation. In addition, CYGB and NGB overexpression upregulated testosterone production. The present study successfully established an inflammatory interaction model of TM3 and RAW264.7 cells. Suppression of CYGB and NGB in TM3 cells changed macrophage morphology, enhanced macrophage cell number and NO release in co-culture experiments upon LPS exposure.In summary, these results demonstrate that globin family members might control LPS induced inflammation by regulating apoptotic mechanisms and macrophage response. |
| |
| Keywords: | Leydig cell Inflammation Lipopolysaccharide Cytoglobin Neuroglobin Testosterone |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
