Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotect...Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotective mechanism of Shuxuetong injection remains poorly understood. Here, cerebral microvascular endothelial cells(bEnd.3) were incubated in glucose-free Dulbecco's modified Eagle's medium containing 95% N_2/5% CO_2 for 6 hours, followed by high-glucose medium containing 95% O_2 and 5% CO_2 for 18 hours to establish an oxygen-glucose deprivation/reperfusion model. This in vitro cell model was administered Shuxuetong injection at 1/32, 1/64, and 1/128 concentrations(diluted 32-, 64-, and 128-times). Cell Counting Kit-8 assay was used to evaluate cell viability. A fluorescence method was used to measure lactate dehydrogenase, and a fluorescence microplate reader used to detect intracellular reactive oxygen species. A fluorescent probe was also used to measure mitochondrial superoxide production. A cell resistance meter was used to measure transepithelial resistance and examine integrity of monolayer cells. The fluorescein isothiocyanate-dextran test was performed to examine blood-brain barrier permeability. Real-time reverse transcription polymerase chain reaction was performed to analyze mRNA expression levels of tumor necrosis factor alpha, interleukin-1β, interleukin-6, and inducible nitric oxide synthase. Western blot assay was performed to analyze expression of caspase-3, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, occludin, vascular endothelial growth factor, cleaved caspase-3, B-cell lymphoma 2, phosphorylated extracellular signal-regulated protein kinase, extracellular signal-regulated protein kinase, nuclear factor-κB p65, I kappa B alpha, phosphorylated I kappa B alpha, I kappa B kinase, phosphorylated I kappa B kinase, claudin-5, and zonula occludens-1. Our results show that Shuxuetong injection increases bEnd.3 cell viability and B-cell lymphoma 2 expression, reduces cleaved caspase-3 expression, inhibits production of reactive oxygen species and mitochondrial superoxide, suppresses expression of tumor necrosis factor alpha, interleukin-1β, interleukin-6, inducible nitric oxide synthase mRNA, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, markedly increases transepithelial resistance, decreases blood-brain barrier permeability, upregulates claudin-5, occludin, and zonula occludens-1 expression, reduces nuclear factor-κB p65 and vascular endothelial growth factor expression, and reduces I kappa B alpha, extracellular signal-regulated protein kinase 1/2, and I kappa B kinase phosphorylation levels. Overall, these findings suggest that Shuxuetong injection has protective effects on brain microvascular endothelial cells after oxygen-glucose deprivation/reperfusion. Moreover, its protective effect is associated with reduction of mitochondrial superoxide production, inhibition of the inflammatory response, and inhibition of vascular endothelial growth factor, extracellular signal-regulated protein kinase 1/2, and the nuclear factor-κB p65 signaling pathway.展开更多
Objective: Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion(I/R)injury,resulting in neuronal death.Salvianolate Lyophilized Inj...Objective: Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion(I/R)injury,resulting in neuronal death.Salvianolate Lyophilized Injection(SLI)and Xueshuantong Injection(Lyophilized)(XST),which have been widely used in the treatment of acutely cerebral infarction clinically in China,exhibit various biological activities.In this study,the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion-reperfusion(MCAO/R)were investigated.Methods: In this study,male Wistar rats were subjected to 1.5 h of middle cerebral artery occlusion followed by reperfusion for 24 h.The rats were randomly divided into the following six groups: normal group(NOR),model group(MOD),SLI group(21 mg/kg,SLI),XST group(100 mg/kg,XST),SLI combined with XST(XST 100 mg/kg + SLI 21 mg/kg,1 X1 S),and Edaravone(as a positive control drug,6 m L/kg,EDI),once a day for 3 d.The neuronal injury,the expression of glial fibrillary acidic protein(GFAP)and ionized calcium binding adaptor molecule 1(IBA-1),and the changes of pro-inflammatory mediators interleukin-6(IL-6),tumor necrosis factor alpha(TNF-α)and anti-inflammatory mediator interleukin-10(IL-10)were observed.Results: 1 X1 S treatment significantly increased the number of neuron,compared with the MOD group,SLI group and XST group.Gliosis(GFAP and IBA-1)and expression of pro-inflammatory mediators IL-6 and TNF-α were significantly reduced.Meanwhile,1 X1 S significantly increased the expression of antiinflammatory mediator IL-10 in the brains of MCAO/R rats,compared with the MOD group,SLI and XST groups.SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group.Conclusions: This study shows that SLI combined with XST(1 X1 S)can protect cerebral ischemiareperfusion injury due to its anti-inflammatory property,and may provide a potential promising new therapeutic strategy for acute ischemic stroke.展开更多
In this work, the polyacrylonitrile containing gadolinium nanofibers for thermal neutron protection were successfully fabricated by electrospunning and followed by in situ self-polymerization. Scanning electron micros...In this work, the polyacrylonitrile containing gadolinium nanofibers for thermal neutron protection were successfully fabricated by electrospunning and followed by in situ self-polymerization. Scanning electron microscopy(SEM) and energy-dispersive spectroscopy(EDS) results show that there are no beads on the smooth surface of the nanofibers and gadolinium elements are uniformly dispersed in the matrix. The thermal analysis and FTIR results prove that gadolinium methacrylate is induced in situ selfpolymerization during the heat treatment. The leaching rate of Gd^(3+) decreases from 79.97% to 10.74% tested by lowfield nuclear magnetic resonance(LF-NMR) method after the self-polymerization of gadolinium methacrylate in the matrix when the nanofibers were immersed in water for7 days. The thermal neutron shielding analysis calculated by MCNP program shows that above 99% thermal neutrons are absorbed when traveling through the 2-mm-thick polyacrylonitrile containing gadolinium nanofibers.展开更多
基金supported in part by the National Natural Science Foundation of China,No.81573644(to LMH),81573733(to SWX)the Tianjin 131 Innovative Team Project,China(to HW)+5 种基金the National Major Science and Technology Project of China,No.2012ZX09101201-004(to SWX)the Science and Technology Plan Project of Tianjin of China,No.16PTSYJC00120(to LMH)the Applied Foundation and Frontier Technology Research Program of Tianjin of China(General Project),No.14JCYBJC28900(to SXW)the National International Science and Technology Cooperation Project of China,No.2015DFA30430(to HW)the Key Program of the Natural Science Foundation of Tianjin of China,No.16ICZDJC36300(to HW)the Scientific Research and Technology Development Plan Project of Guangxi Zhuang Autonomous Region of China,No.14125008-2-5(to SXW)
文摘Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotective mechanism of Shuxuetong injection remains poorly understood. Here, cerebral microvascular endothelial cells(bEnd.3) were incubated in glucose-free Dulbecco's modified Eagle's medium containing 95% N_2/5% CO_2 for 6 hours, followed by high-glucose medium containing 95% O_2 and 5% CO_2 for 18 hours to establish an oxygen-glucose deprivation/reperfusion model. This in vitro cell model was administered Shuxuetong injection at 1/32, 1/64, and 1/128 concentrations(diluted 32-, 64-, and 128-times). Cell Counting Kit-8 assay was used to evaluate cell viability. A fluorescence method was used to measure lactate dehydrogenase, and a fluorescence microplate reader used to detect intracellular reactive oxygen species. A fluorescent probe was also used to measure mitochondrial superoxide production. A cell resistance meter was used to measure transepithelial resistance and examine integrity of monolayer cells. The fluorescein isothiocyanate-dextran test was performed to examine blood-brain barrier permeability. Real-time reverse transcription polymerase chain reaction was performed to analyze mRNA expression levels of tumor necrosis factor alpha, interleukin-1β, interleukin-6, and inducible nitric oxide synthase. Western blot assay was performed to analyze expression of caspase-3, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, occludin, vascular endothelial growth factor, cleaved caspase-3, B-cell lymphoma 2, phosphorylated extracellular signal-regulated protein kinase, extracellular signal-regulated protein kinase, nuclear factor-κB p65, I kappa B alpha, phosphorylated I kappa B alpha, I kappa B kinase, phosphorylated I kappa B kinase, claudin-5, and zonula occludens-1. Our results show that Shuxuetong injection increases bEnd.3 cell viability and B-cell lymphoma 2 expression, reduces cleaved caspase-3 expression, inhibits production of reactive oxygen species and mitochondrial superoxide, suppresses expression of tumor necrosis factor alpha, interleukin-1β, interleukin-6, inducible nitric oxide synthase mRNA, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, markedly increases transepithelial resistance, decreases blood-brain barrier permeability, upregulates claudin-5, occludin, and zonula occludens-1 expression, reduces nuclear factor-κB p65 and vascular endothelial growth factor expression, and reduces I kappa B alpha, extracellular signal-regulated protein kinase 1/2, and I kappa B kinase phosphorylation levels. Overall, these findings suggest that Shuxuetong injection has protective effects on brain microvascular endothelial cells after oxygen-glucose deprivation/reperfusion. Moreover, its protective effect is associated with reduction of mitochondrial superoxide production, inhibition of the inflammatory response, and inhibition of vascular endothelial growth factor, extracellular signal-regulated protein kinase 1/2, and the nuclear factor-κB p65 signaling pathway.
基金financially supported by the National Natural Science Foundation (81573644)the Natural Science Foundation of Tianjin (14JCYBJC28900)the Tianjin Technology Innovation System and the Condition of Platform Construction Plan (16PTSYJC00120)
文摘Objective: Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion(I/R)injury,resulting in neuronal death.Salvianolate Lyophilized Injection(SLI)and Xueshuantong Injection(Lyophilized)(XST),which have been widely used in the treatment of acutely cerebral infarction clinically in China,exhibit various biological activities.In this study,the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion-reperfusion(MCAO/R)were investigated.Methods: In this study,male Wistar rats were subjected to 1.5 h of middle cerebral artery occlusion followed by reperfusion for 24 h.The rats were randomly divided into the following six groups: normal group(NOR),model group(MOD),SLI group(21 mg/kg,SLI),XST group(100 mg/kg,XST),SLI combined with XST(XST 100 mg/kg + SLI 21 mg/kg,1 X1 S),and Edaravone(as a positive control drug,6 m L/kg,EDI),once a day for 3 d.The neuronal injury,the expression of glial fibrillary acidic protein(GFAP)and ionized calcium binding adaptor molecule 1(IBA-1),and the changes of pro-inflammatory mediators interleukin-6(IL-6),tumor necrosis factor alpha(TNF-α)and anti-inflammatory mediator interleukin-10(IL-10)were observed.Results: 1 X1 S treatment significantly increased the number of neuron,compared with the MOD group,SLI group and XST group.Gliosis(GFAP and IBA-1)and expression of pro-inflammatory mediators IL-6 and TNF-α were significantly reduced.Meanwhile,1 X1 S significantly increased the expression of antiinflammatory mediator IL-10 in the brains of MCAO/R rats,compared with the MOD group,SLI and XST groups.SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group.Conclusions: This study shows that SLI combined with XST(1 X1 S)can protect cerebral ischemiareperfusion injury due to its anti-inflammatory property,and may provide a potential promising new therapeutic strategy for acute ischemic stroke.
基金financially supported by Postgraduate Research & Practice Innovation Program of Jiangsu Province (No. KYLX_1337)the Excellent Doctorial Dissertations Fund of Yangzhou University
文摘In this work, the polyacrylonitrile containing gadolinium nanofibers for thermal neutron protection were successfully fabricated by electrospunning and followed by in situ self-polymerization. Scanning electron microscopy(SEM) and energy-dispersive spectroscopy(EDS) results show that there are no beads on the smooth surface of the nanofibers and gadolinium elements are uniformly dispersed in the matrix. The thermal analysis and FTIR results prove that gadolinium methacrylate is induced in situ selfpolymerization during the heat treatment. The leaching rate of Gd^(3+) decreases from 79.97% to 10.74% tested by lowfield nuclear magnetic resonance(LF-NMR) method after the self-polymerization of gadolinium methacrylate in the matrix when the nanofibers were immersed in water for7 days. The thermal neutron shielding analysis calculated by MCNP program shows that above 99% thermal neutrons are absorbed when traveling through the 2-mm-thick polyacrylonitrile containing gadolinium nanofibers.
