Rat astrocytes during anoxia: Secretome profile of cytokines and chemokines

Abstract

IntroductionThe precise mechanisms of the inflammatory responses after cerebral ischemia in vivo are difficult to elucidate because of the complex nature of multiple series of interactions between cells and molecules. This study explored temporal patterns of secretion of 30 cytokines and chemokines from Sprague Dawley rat astrocytes in primary culture in order to elucidate signaling pathways that are triggered by astrocytes during anoxia.MethodsPrimary cultures of rat brain astrocytes were incubated for periods of 2–24 hr in the absence of oxygen (anoxia) or under normal partial pressure of oxygen (controls). Simultaneous detection of 29 cytokines and chemokines in the samples was performed using a rat cytokine array panel, while the temporal pattern of angiopoietin‐1 (Ang‐1) secretion was determined separately using ELISA. Wilcoxon–Mann–Whitney test was used to compare normoxic and anoxic samples and the Hodge–Lehman estimator with exact 95% confidence intervals was computed to assess the size of differences in cytokine secretion. The obtained data were imported into the Core Analysis tool of Ingenuity Pathways Analysis software in order to relate changes in secretion of cytokines and chemokines from astrocytes during anoxia to potential molecular signal networks.ResultsWith the exception of Ang‐1, concentrations of all cytokines/chemokines in samples collected after anoxia exposure were either the same, or higher, than in control groups. No clear pattern of changes could be established for groups of cytokines with similar effects (i.e., pro‐ or anti‐inflammatory cytokines). The pattern of changes in cytokine secretion during anoxia was associated with the HIF‐1α‐mediated response, as well as cytokines IL‐1β and cathepsin S pathways, which are related to initiation of inflammation and antigen presentation, respectively, and to ciliary neurotrophic factor.ConclusionsThese in vitro findings suggest that astrocytes may play a role in triggering inflammation during anoxia/ischemia of the brain.

Hypoxia Alters the Expression of CC Chemokines and CC Chemokine Receptors in a Tumor–A Literature Review

“…In non-cancer cells such as dermal fibroblasts [162] and primary hepatocytes [209] chronic hypoxia does not change the expression of CCL3/MIP-1α and CCL4/MIP-1β. However, in rat brain astrocytes, chronic hypoxia increases CCL3/MIP-1α expression [202]. In NK cells, chronic hypoxia reduces the expression of CCL3/MIP-1α and other pro-inflammatory cytokines [204], which contributes to cancer immunoevasion.…”

“…Nevertheless, some studies show that chronic hypoxia increases, and some that it reduces the expression of this chemokine. CCL5/RANTES expression is increased in acute myeloid leukemia cells [ 185 ], multiple myeloma cells [ 165 ], MDA-MB-231 and MDA-MB-435 breast cancer cells [ 142 , 153 ], cervical squamous carcinoma cells [ 46 ], and primary rat astrocytes [ 202 ].…”

The Effect of Hypoxia on the Expression of CXC Chemokines and CXC Chemokine Receptors—A Review of Literature

“…It can increase the expression of a protein or miRNA which directly regulates the expression of this chemokine. In contrast, chronic hypoxia reduces the expression of CXCL3 in primary rat astrocytes [ 165 ] and SK-OV-3 ovarian adenocarcinoma cells [ 14 ].…”

“…Chronic hypoxia increases CXCL7 expression in cervical cancer cells [ 169 ] and primary rat astrocytes [ 165 ]. The overexpression of HIF-2α increases the expression of this chemokine in murine articular chondrocytes [ 163 ].…”

Gliome database: a comprehensive web-based tool to access and analyze glia secretome data

“…Secretome is a term that denotes all the factors secreted by cells, such as soluble proteins, cytokines ( 10 ), growth factors ( 11 ), extracellular matrix proteins ( 12 ) and shed membrane receptors ( 13 ). The secretory proteins typically include a hydrophobic signal peptide at the N -terminus undergo their journey along the biosynthetic-secretory pathway through the endoplasmic reticulum and Golgi apparatus.…”