Role of high-mobility group box 1 in methamphetamine-induced activation and migration of astrocytes.
BACKGROUND
Mounting evidence has indicated that high-mobility group box 1 (HMGB1) is involved in cell activation and migration. Our previous study demonstrated that methamphetamine mediates activation of astrocytes via sigma-1 receptor (σ-1R). However, the elements downstream of σ-1R in this process remain poorly understood. Thus, we examined the molecular mechanisms involved in astrocyte activation and migration induced by methamphetamine.


METHODS
The expression of HMGB1, σ-1R, and glial fibrillary acidic protein (GFAP) was examined by western blot and immunofluorescent staining. The phosphorylation of cell signaling pathways was detected by western blot, and cell migration was examined using a wound-healing assay in rat C6 astroglia-like cells transfected with lentivirus containing red fluorescent protein (LV-RFP) as well as in primary human astrocytes. The role of HMGB1 in astrocyte activation and migration was validated using a siRNA approach.


RESULTS
Exposure of C6 cells to methamphetamine increased the expression of HMGB1 via the activation of σ-1R, Src, ERK mitogen-activated protein kinase, and downstream NF-κB p65 pathways. Moreover, methamphetamine treatment resulted in increased cell activation and migration in C6 cells and primary human astrocytes. Knockdown of HMGB1 in astrocytes transfected with HMGB1 siRNA attenuated the increased cell activation and migration induced by methamphetamine, thereby implicating the role of HMGB1 in the activation and migration of C6 cells and primary human astrocytes.


CONCLUSIONS
This study demonstrated that methamphetamine-mediated activation and migration of astrocytes involved HMGB1 up-regulation through an autocrine mechanism. Targeting HMGB1 could provide insights into the development of a potential therapeutic approach for alleviation of cell activation and migration of astrocytes induced by methamphetamine.
Department of Pharmacology, Medical School of Southeast University, Nanjing, 210009, China.
Yuan
Y
Zhang
Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China.
Tiebing
T
Zhu
Department of Pharmacology, Medical School of Southeast University, Nanjing, 210009, China.
Xiaotian
X
Zhang
Department of Physiology, Medical School of Southeast University, Nanjing, China.
Jie
J
Chao
Department of Pharmacology, Nanjing Medical University, Nanjing, China.
Gang
G
Hu
Department of Pharmacology, Medical School of Southeast University, Nanjing, 210009, China. yaohh@seu.edu.cn.|Institute of Life Sciences, Key Laboratory of Developmental Genes and Human Disease, Southeast University, Nanjing, Jiangsu, China. yaohh@seu.edu.cn.
Honghong
H
Yao
D000818:Animals; D001253:Astrocytes / Q000187:drug effects*; D002070:Butadienes / Q000494:pharmacology; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000187:drug effects*; D015259:Dopamine Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005029:Ethylenediamines / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D024243:HMGB1 Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008694:Methamphetamine / Q000494:pharmacology*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors
D002070:Butadienes; D015259:Dopamine Agents; D004791:Enzyme Inhibitors; D005029:Ethylenediamines; D005904:Glial Fibrillary Acidic Protein; D024243:HMGB1 Protein; D016328:NF-kappa B; D009570:Nitriles; D034741:RNA, Small Interfering; C113580:U 0126; C097963:N-(2-(3,4-Dichlorphenyl)ethyl)-N,N',N'-trimethyl-1,2-ethandiamin; D008694:Methamphetamine; D020929:Mitogen-Activated Protein Kinase Kinases