Impaired wound healing after cerebral hypoxia-ischemia in the diabetic mouse.
Impaired peripheral wound healing is a hallmark of diabetics pathology and has been attributed to compromised macrophage activation. Stroke is another component of diabetic pathology, with increased tissue infarction and worsened recovery although the mechanisms remain unresolved. In this study, we investigated whether a compromised glial/macrophage response might contribute to cerebral hypoxic-ischemic (H/I) brain damage in diabetic (db/db), relative to their normoglycemic db/+ mice. Hypoxia-ischemia was induced in 8-week-old male db/db and db/+ mice by the ligation of right common carotid artery followed by systemic hypoxia (8% O2: 92% N2) for 17 mins. Mice were killed at specific intervals of reperfusion/recovery and the brains analyzed by in situ hybridization or total RNA isolation. In situ hybridization using bfl-1 (microglia) and glial fibrillary acidic protein (GFAP) (astrocytes) revealed expression of both bfl-1 and GFAP in the ipsilateral hemisphere at 4 h in the db/+ mice, which was delayed and minimal in the db/db mice. RNase protection assays showed a robust increase in expression of the proinflammatory cytokines tumor necrosis factor-alpha (TNFalpha), interleukin-1 IL-1alpha, and IL-1beta mRNA in the db/+ mice at 6 to 8 h of reperfusion peaking at 8 to 12 h; in db/db mice expression was markedly delayed and diminished. Real-time-polymerase chain reaction (RT-PCR) confirmed the reduced and delayed expression TNFalpha, IL-1alpha, IL-1beta, and the growth factors insulin-like growth factor-1 and ciliary neurotrophic factor in the db/db mice; enzyme-linked immunosorbent assays confirmed the reduced and delayed translation of IL-1beta protein. These findings suggest that a compromised inflammatory response may underlie the greater infarct associated with diabetic db/db mice compared with their nondiabetic littermates following a hypoxic/ischemic insult.
Kumari
Rashmi
R
Department of Neural and Behavioral Sciences, Milton S Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Willing
Lisa B
LB
Krady
J Kyle
JK
Vannucci
Susan J
SJ
Simpson
Ian A
IA
D000818:Animals; D001253:Astrocytes / Q000502:physiology; D020934:Ciliary Neurotrophic Factor / Q000096:biosynthesis; D003920:Diabetes Mellitus / Q000235:genetics* / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D020925:Hypoxia-Ischemia, Brain / Q000473:pathology*; D017403:In Situ Hybridization; D007334:Insulin-Like Growth Factor I / Q000096:biosynthesis; D053582:Interleukin-1alpha / Q000096:biosynthesis / Q000502:physiology; D053583:Interleukin-1beta / Q000096:biosynthesis / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000502:physiology; D015778:Minor Histocompatibility Antigens; D023282:Nuclease Protection Assays; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D014945:Wound Healing / Q000502:physiology*
C096816:BCL2-related protein A1; D020934:Ciliary Neurotrophic Factor; D005904:Glial Fibrillary Acidic Protein; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D015778:Minor Histocompatibility Antigens; D019253:Proto-Oncogene Proteins c-bcl-2; D014409:Tumor Necrosis Factor-alpha; D007334:Insulin-Like Growth Factor I