title: G-CSF protects human brain vascular endothelial cells injury induced by high glucose, free fatty acids and hypoxia through MAPK and Akt signaling.
abstract: Granulocyte-colony stimulating factor (G-CSF) has been shown to play a neuroprotective role in ischemic stroke by mobilizing bone marrow (BM)-derived endothelial progenitor cells (EPCs), promoting angiogenesis, and inhibiting apoptosis. Impairments in mobilization and function of the BM-derived EPCs have previously been reported in animal and human studies of diabetes where there is both reduction in the levels of the BM-derived EPCs and its ability to promote angiogenesis. This is hypothesized to account for the pathogenesis of diabetic vascular complications such as stroke. Here, we sought to investigate the effects of G-CSF on diabetes-associated cerebral vascular defect. We observed that pretreatment of the cultured human brain vascular endothelial cells (HBVECs) with G-CSF largely prevented cell death induced by the combination stimulus with high glucose, free fatty acids (FFA) and hypoxia by increasing cell viability, decreasing apoptosis and caspase-3 activity. Cell ultrastructure measured by transmission electron microscope (TEM) revealed that G-CSF treatment nicely reduced combination stimulus-induced cell apoptosis. The results from fluorescent probe Fluo-3/AM showed that G-CSF greatly suppressed the levels of intracellular calcium ions under combination stimulus. We also found that G-CSF enhanced the expression of cell cycle proteins such as human cell division cycle protein 14A (hCdc14A), cyclinB and cyclinE, inhibited p53 activity, and facilitated cell cycle progression following combination stimulus. In addition, activation of extracellular signal-regulated kinase1/2 (ERK1/2) and Akt, and deactivation of c-Jun N terminal kinase (JNK) and p38 were proved to be required for the pro-survival effects of G-CSF on HBVECs exposed to combination stimulus. Overall, G-CSF is capable of alleviating HBVECs injury triggered by the combination administration with high glucose, FFA and hypoxia involving the mitogen-activated protein kinases (MAPK) and Akt signaling cascades. G-CSF may represent a promising therapeutic agent for diabetic stroke.
authors: Department of Neurology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, Shanghai, 200011, China.
authors: Jingjing
authors: J
authors: Su
authors: Department of Geriatric Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Houguang
authors: H
authors: Zhou
authors: Department of General Medicine, Ouyang Hospital, Hongkou District, Shanghai, China.
authors: Yinghong
authors: Y
authors: Tao
authors: State Key Laboratory of Medical Neurobiology, Department of Neurobiology, School of Basic Medical Science, Shanghai Medical College, Fudan University, Shanghai, 200032,China.
authors: Jingchun
authors: J
authors: Guo
authors: Department of Emergency Neurology, the Affiliated Hospital of Medical College Qingdao University, Qingdao, 266100, China.
authors: Zhuangli
authors: Z
authors: Guo
authors: Department of Endocrine, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Shuo
authors: S
authors: Zhang
authors: Department of Geriatric Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Yu
authors: Y
authors: Zhang
authors: Department of Geriatric Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Yanyan
authors: Y
authors: Huang
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Yuping
authors: Y
authors: Tang
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Qiang
authors: Q
authors: Dong
authors: Department of Endocrine, Huashan Hospital, Fudan University, Shanghai, 200040, China.
authors: Renming
authors: R
authors: Hu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D001930:Brain Injuries / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005230:Fatty Acids, Nonesterified / Q000009:adverse effects*; D005947:Glucose / Q000009:adverse effects*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D000860:Hypoxia / Q000503:physiopathology*; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D005230:Fatty Acids, Nonesterified; D020011:Protective Agents; D012333:RNA, Messenger; D016179:Granulocyte Colony-Stimulating Factor; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases; D005947:Glucose