ABSTRACT
@#Objective To investigate the effects and related molecular mechanisms of glutathione S-transferase pi 1 (GSTP1) on the inflammasome activation of astrocytes.Methods A model of epilepsy was established in 10-week-old male SD rats by intraperitoneal injection of lithium chloride (n=8),and brain tissues were collected from the hippocampus.Rat primary astrocytes were treated with different concentrations of lipopolysaccharide (0 μg/ml,0.1 μg/ml,1 μg/ml,10 μg/ml and 100 μg/ml) for 48 h.The protein levels of GSTP1,JNK and p-JNK in tissues and cells were measured by Western blotting.The levels of TNF-α,IL-1β,IL-6 were detected by enzyme-linked immunosorbent assay.The levels of glutamate (Glu) were detected by high pressure liquid chromatography.The lipopolysaccharide-induced astrocytes were transiently transfected with GSTP1 overexpression vector and were treated with Anisomycin (JNK activator),and the inflammatory activation of astrocytes was observed.Results The protein levels of GSTP1 were lower in the hippocampal brain tissues of epileptic rats than those in normal rats,while the protein levels of p-JNK and the levels of TNF-α,IL-1β,IL-6 and Glu were higher in epileptic rats than those in normal rats (P<0.05).GSTP1 was negatively correlated with p-JNK protein expression level (P<0.05).Lipopolysaccharide-induced inflammasome activation in astrocytes,as evidenced by a dose-dependent decrease in the protein expression levels of GSTP1 and a dose-dependent increase in the protein levels of p-JNK and the levels of TNF-α,IL-1β,IL-6 and Glu (P<0.05).Overexpression of GSTP1 inhibited lipopolysaccharide-induced inflammasome activation of astrocytes,while Anisomycin partially reversed the inhibitory effect of GSTP1. Conclusion GSTP1 inhibits inflammasome activation of astrocytes in the hippocampus of epileptic rats,and its molecular mechanism is related to JNK pathway inhibition.
ABSTRACT
@#Objective To investigate the role and molecular mechanism of long intergenic non-coding RNA 1116 (LINC01116) in hippocampal astrocytes of acute ischemic stroke (AIS).Methods The expressions of LINC01116 and miR-203 in serum of 131 AIS patients before and after thrombolysis were detected by real-time fluorescence quantitative PCR.The regulatory effect of LINC01116 on miR-203 was detected by dual-luciferase reporter gene and RNA-binding protein immunoprecipitation assay.Human hippocampal astrocytes (hHA) were applied to establish an oxygen-glucose deprivation/reoxygenation (OGD/R) model and were treated with LINC01116 interference and LINC01116 combined with miR-203 interference.The changes of cell proliferation,cell apoptosis,production of reactive oxygen species (ROS),activity of lactate dehydrogenase (LDH),and inflammatory factors were detected by CCK-8,TUNEL and Western blotting,ROS assay,LDH assay,and enzyme-linked immunosorbent assay,respectively.Results The expressions of LINC01116 and miR-203 in serum after thrombolysis were higher and lower than those before thrombolysis,respectively,and the expressions of LINC01116 and miR-203 were negatively correlated (P<0.05).LINC01116 inhibited miR-203 expression by sponge of miR-203 (P<0.05).LINC01116 interference alleviated the OGD/R-induced injury of hHA cells,which manifested as elevated cell proliferation ability,decreased cell apoptosis rate,decreased protein expressions of cleaved caspase-3 and Bax but raised protein expression of Bcl-2,reduced ROS production,decreased LDH activity,and downregulated TNF-α,IL-1β and IL-6 concentrations but upregulated IL-4,IL-10 and IL-13 concentrations (P<0.05).miR-203 interference reversed the protective effect of LINC01116 interference on the OGD/R-induced injury of hHA cells (P<0.05).Conclusions LINC01116 promotes the OGD/R-induced injury of hHA cells by targeting miR-203,suggesting that the LINC01116/miR-203 pathway might be a potential therapeutic target for AIS.