Knockdown of IL-1β Improves Hypoxia–ischemia Brain Associated with IL-6 Up-regulation in Cell and Animal Models

Abstract A study was conducted to investigate the effect of interleukin-1β (IL-1β) on hypoxia ischemia (HI) of cultured astrocyte and neonatal rat models and to explore the underlying molecular regulation mechanism. Primary rat astrocyte was exposed to hypoxia (2 % O2, 98 % N2) and cultured in serum-free medium for 6, 12, and 18 h to establish cell model of HI. Morphologic changes of astrocyte were monitored and gene expression change of IL-1β evaluated by real-time polymerase chain reaction (PCR). To establish the HI animal model, 3 days old postnatal Sprague–Dawley (SD) rats were treated with the right carotid artery ligation and were exposed to 8 % oxygen for 8, 16 and 24 h, respectively. Longa score scale, hematoxylin and eosin (HE) staining and water content were examined to assess neurologic function and morphology changes. IL-1β siRNA lentivirus (IL-1β-RNAi-LV) was injected into cerebral cortex motor area 2 days before HI and the interference efficiency examined by real-time PCR and Western blotting, respectively. Immunofluorescence staining of GFAP and IL-1β was performed to identify the location and interference effect of IL-1β, respectively. To further explore the potential mechanisms, the expression of inflammatory factors, including IL-6, IL-10 and tumor necrosis factor-alpha (TNF-α), was examined following IL-1β down-regulation. The size of soma astrocyte was increased greatly after 12 and 18 h of HI with IL-1β up-regulati...
Source: Molecular Neurobiology - Category: Neurology Source Type: research