Effects of autophagy inhibition on the inflammatory response in cerebral microvascular endothelial cells following cerebral ischemia-reperfusion injury under hyperglycemic conditions in rats
HU Jun-hua
LI Ling-ling
LI Ya-tan
YANG Pei-e
AISHA Abdulla
YUAN Yang
ZHAO Jia-le
WU Qin-fen
GULIBAHA Maimaitili
Abstract：Objective To investigate the role of autophagy in cerebral microvascular endothelial cell inflammation following cerebral ischemia-reperfusion injury(CI/RI)under hyperglycemic conditions and its molecular mechanism.Methods Ninety male SD rats with streptozotocin(STZ)-induced diabetes were randomly divided into three groups:Sham group(n=30),CI/RI group(n=30),and autophagy inhibitor group(n=30).CI/RI models were established using the intraluminal filament method,with 3-MA administered intraperitoneally to inhibit autophagy.Cerebral infarct volume was quantified by TTC staining,neurological deficits were assessed via Garcia JH scores,blood-brain barrier permeability was evaluated by Evans Blue extravasation,histopathological changes were observed by HE staining,serum IL-6,IL-1β,and TNF-α levels were measured by ELISA,and NLRP3,IL-1β,and TNF-α protein expression in brain tissue was analyzed by Western blotting.Primary cerebral microvascular endothelial cells were cultured and subjected to oxygen-glucose deprivation/reoxygenation,followed by Western blotting to assess LC3-II/LC3-I ratio,Beclin1,P62,and NLRP3 inflammasome-related protein expression.Results Compared with the CI/RI group,the 3-MA group showed significantly reduced cerebral infarct volume(P<0.05),improved neurological function scores(P<0.05),decreased Evans Blue leakage(P<0.05),and attenuated histopathological damage(P<0.05).3-MA intervention markedly lowered serum IL-6,IL-1β,and TNF-α levels(P<0.05)and suppressed NLRP3,IL-1β,and TNF-α protein expression in brain tissue(P<0.05).In vitro experiments demonstrated that 3-MA decreased the LC3-II/LC3-I ratio and Beclin1 expression,increased P62 accumulation,and inhibited NLRP3 inflammasome activation along with downstream inflammatory cytokine expression(P<0.05).Conclusion The autophagy inhibitor 3-MA alleviates cerebral microvascular endothelial cell inflammation and improves neurological outcomes in diabetic rats after CI/RI,likely through suppressing NLRP3 inflammasome activation.This study suggests that targeting autophagy may represent a therapeutic strategy for diabetes-associated cerebral ischemia-reperfusion injury.
Keywords：Ischemia-reperfusion injuryDiabetesAutophagyInflammatory responseBrain microvascular endothelial cells
Publication Date：2025-04-25
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 228-234 )