Objective To investigate the role and underlying mechanism of spermidine(SPD)in intracerebral hemorrhage(ICH).Methods Male C57BL/6 mice were subjected to establish a collagenase-induced ICH model.The 108 mice were randomly divided into Sham group,ICH group and ICH+SPD group(intraperitoneal injection of 15 mg/kg SPD for 3 consecutive days after modeling),with 36 mice in each group.On the 3rd day after ICH,neurological deficits were evaluated using modified Garcia scoring and forelimb placing test;brain pathological damage was assessed with HE staining;activation of microglia/macrophages(Iba-1)and astrocytes(GFAP)was detected by immunofluorescence assay;expression of autophagy markers(Beclin-1,P62)and inflammatory factors(MMP-9,NLRP3,COX-2)was measured with Western blotting.In in vitro experiments,hemin was used to stimulate HT-22 cells to mimic ICH.The HT-22 cells were randomly divided into Control group,Hemin group,Hemin+SPD group,and Hemin+SPD+3-methyladenine(3-MA,an autophagy inhibitor)group(n=7).After 24 h of hemin treatment,cell viability was detected with CCK-8 assay,the expression of autophagy-related proteins(LC3-Ⅱ and P62)were detected with Western blotting,and oxidative stress was determined by measuring superoxide dismutase(SOD)activity and malondialdehyde(MDA)content.Results On day 3 post-ICH,SPD significantly reduced the area of brain damage(P<0.05),improved neurological recovery(P<0.05),activated autophagy with up-regulation of Beclin-1 while down-regulation of P62(P<0.05),suppressed the activation of microglia/macrophage and astrocytes(P<0.01),reduced the expression of MMP-9,NLRP3 and COX-2,and enhanced SOD activity and decreased MDA content(P<0.05)when compared with the ICH group.SPD increased the viability of HT-22 cells(P<0.05),improved SOD activity and reduced MDA content(P<0.01).Autophagy inhibitor 3-MA effectively blocked the down-regulation of LC3-Ⅱ and up-regulation of P62,and completely reversed above protective effects caused by SPD(P<0.05).Conclusion SPD activates autophagy after ICH and improves post-ICH brain injury by suppressing neuroinflammation and oxidative stress.

Objective To explore the biological behavior and mechanism of isovitexin(Isov)on pancreatic cancer cells.Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines,and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50).The PC cell line PANC-1 cells were grouped into the control group,the Isov group,the Isov+in-miR-NC group,the Isov+in-miR-339-5p group,the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group.The survival,migration and invasion of PANC-1 cells were detected by CCK-8,scratch healing assay and Transwell assay.Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8(HSPA8)in PANC-1 cells.Western blot assay was used to detect protein HSPA8 expression in various groups of cells.Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8.A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov.Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells.Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells,reduce the number of invasive cells,up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels(P＜0.05),while these effects were blocked by down-regulated miR-339-5p(P＜0.05).In addition,HSPA8 was the target gene of miR-339-5p,and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells.In vivo studies confirmed that after Isov treatment,the tumor volume and weight of nude mice decreased,the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased(P＜0.05).Conclusion Isov may inhibit the proliferation,migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.

Objective To explore the biological behavior and mechanism of isovitexin(Isov)on pancreatic cancer cells.Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines,and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50).The PC cell line PANC-1 cells were grouped into the control group,the Isov group,the Isov+in-miR-NC group,the Isov+in-miR-339-5p group,the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group.The survival,migration and invasion of PANC-1 cells were detected by CCK-8,scratch healing assay and Transwell assay.Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8(HSPA8)in PANC-1 cells.Western blot assay was used to detect protein HSPA8 expression in various groups of cells.Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8.A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov.Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells.Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells,reduce the number of invasive cells,up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels(P＜0.05),while these effects were blocked by down-regulated miR-339-5p(P＜0.05).In addition,HSPA8 was the target gene of miR-339-5p,and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells.In vivo studies confirmed that after Isov treatment,the tumor volume and weight of nude mice decreased,the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased(P＜0.05).Conclusion Isov may inhibit the proliferation,migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.