AIM: To clarify the modulation of autophagy in ischemic penumbra by hydroxysafflor yellow A ( HSYA) after cerebral ischemia/reperfusion ( I/R) injury.METHODS:Male SD rats subjected to transient middle cere-bral artery occlusion for 90 min were randomly divided into 4 groups:sham-operation (sham) group, cerebral I/R (I/R) group, I/R+HSYA group and sham+HSYA group.Modified neurological severity score (mNSS) was used to evaluate the neurological deficits of the rats at 6 h, 1 d, 3 d and 7 d after reperfusion , accompanied by the detection of autophagy , apoptosis and mRNA expression of IFN-βin ischemic penumbra .RESULTS:Compared with sham group , upregulation of LC3-Ⅱand degradation of SQSTM1/P62 were observed in I/R group, indicating the activation of autophagy after I/R.The activation of autophagy in I/R+HSYA group was significantly enhanced by HSYA on day 1 and day 3, and inhibited on day 7, as compared with I/R group (P<0.05).Besides, the mRNA expression of IFN-βin I/R group was significantly upregulated at 6 h, then downregulated on day 1 and day 3, and returned to basal level on day 7, as compared with sham group (P<0.05).In I/R+HSYA group, the mRNA expression of IFN-βwas significantly upregulated on day 1 and day 3, accompanied by the inhibition of apoptosis on day 3 and the significantly decreased mNSS from day 4, as compared with I/R group (P<0.05).CONCLUSION:HSYA alleviates cerebral I/R injury by dynamically modulating the activation of autophagy and the expression of IFN-βin ischemic penumbra .

AIM:To observe the Toll-like receptor 9 (TLR9) activation in microglia BV-2 cells after oxygen-glucose deprivation and reoxygenation ( OGDR) , and its effects on neuronal apoptosis.METHODS:The BV-2 cell super-natants were collected after the corresponding treatment and added to mouse primary cortical neurons after OGDR for 4 h, followed by normal culture for 24 h.The cells were divided into normal BV-2 group, NC-siRNA group, TLR9-siRNA group, OGDR group, OGDR+NC-siRNA group, OGDR+TLR9-siRNA group and control group (without adding BV-2 cell supernatant) .The changes of the neuronal morphology were observed under an inverted phase-contrast microscope, and the neuronal apoptosis was detected by TUNEL.The protein expression of cleaved caspase-3 was detected by Western blot-ting.RESULTS:After OGDR, the axon turned thin, twisted and broken, and neuronal swelling, decrease in refraction and vacuolar degeneration were observed.The green-stained apoptotic bodies in the neurons in all groups were positive. Compared with control group, the caspase-3 protein levels in other groups were increased.Compared with the normal BV-2 group, the caspase-3 protein in OGDR group and TLR9-siRNA group was increased.Compared with OGDR+TLR9-siRNA group, the caspase-3 protein in TLR9-siRNA group and OGDR group was decreased.CONCLUSION: After OGDR, TLR9 activation in BV-2 cells induces neuronal apoptosis with the increase in caspase-3 protein level.Inhibition of TLR9 expression reduces neuronal damage.

Objective To investigate the associations of Toll-like receptors (TLRs) mRNA expressions and their downstream signaling pathways in peripheral blood mononuclear cells with clinical outcomes in patients with cerebral infarction.Methods Ninety patients with cerebral infarction of anterior circulation,admitted to our hospital fiom May 2015 to December 2015,were chosen.Quantitative real-time PCR was performed to measure the expressions ofTLR2,TLR4,TLR3,TLR7,TLR8 and TLR9 and downstream signaling molecules Toll-like receptors related molecule (TRAM),interferon regulatory factor 3 (IRF-3),and interferon beta (IFN-β) in peripheral blood mononuclear cells.Good functional outcome was defined as modified Rankin Scale (mRS) scores≤ 2 90 d after onset.National Institutes of Health Stroke Scale (NIHSS) on admission was used to evaluate the severity of stroke,and infarct volume 7-14 d after onset was measured on MR imaging.The baseline characteristics (TLRs,population information,risk factors,stroke types,clinical and imaging data) were compared between patients with good and bad functional outcomes.Multi-factor Logistics regression analysis was performed.Results TLR3 and TLR7 mRNA expressions were correlated with good functional outcome at acute cerebral infarction,and only TLR3 expression was correlated with good functional outcome at sub-acute phase.TLR3 mRNA expression,C-reactive protein (CRP) level,and NIHSS scores on admission were positively correlated with good outcome (OR=4.435,P=0.001;OR=l.12,P=0.033;OR=1.961,P=0.000).The expressions of TLR3 and IRF-3 were both inversely correlated with NIHSS scores and cerebral infarction volume;IFN-β mRNA expression was positively correlated to TLR3 expression (r=0.392,P=0.000);IFN-β expression and IRF level were positively correlated (r=0.347,P=0.01).Conclusion The TLR3 expression is correlated with good functional outcome in peripheral blood mononuclear cells of patients with cerebral infarction,which may play a crucial role in the survival of neurons after ischemia through spuring the extraction of anti-inflammatory factor IFN-β.

Brain Ischemia ; pathology ; physiopathology ; Humans ; Neoplasm Proteins ; metabolism ; Pyroptosis ; physiology ; Signal Transduction ; physiology

Acyl-CoA synthetase long chain family member 5 (ACSL5), is a member of the acyl-CoA synthetases (ACSs) family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs. The dysregulation of ACSL5 has been reported in some cancers, such as glioma and colon cancers. However, little is known about the role of ACSL5 in acute myeloid leukemia (AML). We found that the expression of ACSL5 was higher in bone marrow cells from AML patients compared with that from healthy donors. ACSL5 level could serve as an independent prognostic predictor of the overall survival of AML patients. In AML cells, the ACSL5 knockdown inhibited cell growth both in vitro and in vivo. Mechanistically, the knockdown of ACSL5 suppressed the activation of the Wnt/β-catenin pathway by suppressing the palmitoylation modification of Wnt3a. Additionally, triacsin c, a pan-ACS family inhibitor, inhibited cell growth and robustly induced cell apoptosis when combined with ABT-199, the FDA approved BCL-2 inhibitor for AML therapy. Our results indicate that ACSL5 is a potential prognosis marker for AML and a promising pharmacological target for the treatment of molecularly stratified AML.
Humans ; Antineoplastic Agents/therapeutic use* ; Apoptosis ; beta Catenin/metabolism* ; Biomarkers, Tumor/metabolism* ; Cell Line, Tumor ; Coenzyme A Ligases/metabolism* ; Leukemia, Myeloid, Acute/metabolism* ; Lipoylation ; Prognosis ; Wnt Signaling Pathway