title: 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-regulation. IL-1β knockdown by siRNA in vitro or by lentivirus in vivo can reverse cell swelling, brain edema and neurologic function deficiencies induced by HI. Lastly, interference of IL-1β remarkably increased IL-6 expression but not IL-10 and TNF-α. Therefore, down-regulation of IL-1β improves the deficiencies of neurologic function and morphology induced by HI, maybe closely associating with IL-6 regulation.
authors: Shenzhen Children's Hospital, Shenzhen, Guangdong, 518038, China.
authors: Sujuan
authors: S
authors: Liu
authors: Shengyun
authors: S
authors: Zhu
authors: Yu
authors: Y
authors: Zou
authors: Tinghua
authors: T
authors: Wang
authors: Xuemei
authors: X
authors: Fu
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004195:Disease Models, Animal; D055785:Gene Knockdown Techniques / Q000379:methods; D020925:Hypoxia-Ischemia, Brain / Q000235:genetics / Q000378:metabolism*; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D015850:Interleukin-6 / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation / Q000502:physiology*
chemical_list: D053583:Interleukin-1beta; D015850:Interleukin-6