目的探究MicroRNA-124(miR-124)对脑缺血损伤小鼠神经功能的改善作用及其内在机制。方法①取32只C57小鼠随机分为假手术组和脑缺血组(n=16),TTC染色和神经行为学评分检测小鼠脑损伤后3天脑梗死面积和运动情况,RT-qPCR检测miR-124在两组...目的探究MicroRNA-124(miR-124)对脑缺血损伤小鼠神经功能的改善作用及其内在机制。方法①取32只C57小鼠随机分为假手术组和脑缺血组(n=16),TTC染色和神经行为学评分检测小鼠脑损伤后3天脑梗死面积和运动情况,RT-qPCR检测miR-124在两组间的表达变化。②培养原代神经元细胞,通过氧糖剥夺模拟缺血再灌注损伤,观察氧糖剥夺后0、72 h miR-124的变化趋势。③另取32只C57小鼠随机分为脑缺血组和脑缺血+miR-124组(n=16),TTC染色、转棒实验和肢体放置实验检测小鼠的运动功能改善情况。④细胞实验分为氧糖剥夺组和氧糖剥夺+miR-124组,通过免疫荧光实验检测两组神经元轴突的生长情况。Targetscan数据库预测miR-124下游靶基因,双荧光素酶基因报告和Western blot在原代神经元等上验证对照组和miR-124组下游靶基因第10号染色体缺失的磷酸酶及张力蛋白同源基因(the phosphate and tensin homolog deleted on chromosome ten,PTEN)的表达变化。上调PTEN,检测氧糖剥夺+miR-124和氧糖剥夺+miR-124+PTEN两组神经元轴突的生长情况。(5)在体实验中检测脑缺血+miR-124组和脑缺血+miR-124+Pten组(n=8)转棒实验和肢体放置情况。结果脑缺血再灌注后,小鼠脑梗死面积约达到40%(P<0.001),小鼠的神经认知功能显著降低(P<0.001),已达到重度认知功能损害;miR-124的表达较对照组明显升高(P<0.05);而进一步过表达miR-124能够促进氧糖剥夺后神经元轴突的生长(P<0.05),显著缩小脑梗死面积(P<0.05),且促进动物运动能力的恢复和改善(P<0.05);Targetscan数据库预测miR-124下游靶基因为PTEN,双荧光素酶基因报告和Western blot都证实miR-124能有效下调PTEN的表达(P<0.05);最后,上调PTEN逆转了miR-124促进神经元轴突的生长以及改善小鼠运动能力的效应(P<0.05)。结论miR-124通过下调PTEN促进脑缺血小鼠神经元轴突生长,且有助于小鼠运动功能的恢复。展开更多
目的:探讨白藜芦醇(RES)预处理对氧糖剥夺(OGD)诱导的大鼠神经干细胞(NSCs)损伤的影响。方法:取孕15 d SD大鼠体内胎鼠脑组织原代培养NSCs,实验分为4组,分别为对照组、OGD模型组、RES+OGD处理组和RES+OGD+sirtinol处理组。利用CCK-8试...目的:探讨白藜芦醇(RES)预处理对氧糖剥夺(OGD)诱导的大鼠神经干细胞(NSCs)损伤的影响。方法:取孕15 d SD大鼠体内胎鼠脑组织原代培养NSCs,实验分为4组,分别为对照组、OGD模型组、RES+OGD处理组和RES+OGD+sirtinol处理组。利用CCK-8试剂盒检测各组NSCs的细胞活性,利用乳酸脱氢酶(LDH)检测各组NSCs的损伤程度。采用Western Blot检测各组NSCs中沉默信息调节因子1(SIRT1)和解偶联蛋白2(UCP2)的蛋白表达水平。结果:OGD损伤能够降低NSCs中SIRT1蛋白表达(P<0.05),RES预处理能够上调OGD损伤后NSCs中SIRT1蛋白的表达(P<0.05);RES预处理能够提高OGD损伤模型中NSCs的细胞活性(P<0.05),降低LDH漏出量(P<0.05),下调NSCs中UCP2的蛋白表达。SIRT1抑制剂sirtinol能够降低OGD损伤后NSCs的细胞活性(P<0.05),增加LDH漏出量(P<0.05),上调NSCs中UCP2的蛋白表达(P<0.05)。结论:RES预处理对OGD诱导的大鼠NSCs损伤具有保护作用,其作用机制可能与SIRT1-UCP2通路有关。展开更多
Objective:To explore the effects of dopamine receptor D2(DRD2)on astrocytic dedifferentiation based on SOX2-regulated genes in neural stem cells(NSCs)and astrocytes.Methods:Immunofluorescence staining and SOX2-GFP mic...Objective:To explore the effects of dopamine receptor D2(DRD2)on astrocytic dedifferentiation based on SOX2-regulated genes in neural stem cells(NSCs)and astrocytes.Methods:Immunofluorescence staining and SOX2-GFP mice were used to examine the lineage differentiation of SOX2-positive cells during the development of cerebral cortex.Primary NSCs/astrocytes culture,ChIP-seq and Western Blot were adopted to analyze and verify the expression of candidate genes.Pharmacological manipulation,neurosphere formation,photochemical ischemia,immunofluorescence staining and behavior tests were adopted to evaluate the effects of activating DRD2 signaling on astrocytic dedifferentiation.Results:Immunofluorescence staining demonstrated the NSC-astrocyte switch of SOX2-expression in the normal development of cerebral cortex.ChIP-seq revealed enrichment of DRD2 signaling by SOX2-bound enhancers in NSCs and SOX2-bound promoters in astrocytes.Western Blot and immunofluorescence staining verified the expression of DRD2 in NSCs and reactive astrocytes.Application of quinagolide hydrocholoride(QH),an agonist of DRD2,significantly promoted astrocytic dedifferentiation both in vitro and in vivo following ischemia.In addition,quinagolide hydrocholoride treatment improved locomotion recovery.Conclusion:Activating DRD2 signaling facilitates astrocytic dedifferentiation and may be used to treat ischemic stroke.展开更多
目的观察双LVIS支架套叠辅助弹簧圈栓塞治疗颅内血泡样动脉瘤(BBA)的效果。方法回顾性分析45例接受支架辅助弹簧圈栓塞治疗的BBA患者资料,按治疗方式分为双LVIS支架组(DLS组,18例)和非双LVIS支架组(NDLS组,27例),对比2组手术效果、围手...目的观察双LVIS支架套叠辅助弹簧圈栓塞治疗颅内血泡样动脉瘤(BBA)的效果。方法回顾性分析45例接受支架辅助弹簧圈栓塞治疗的BBA患者资料,按治疗方式分为双LVIS支架组(DLS组,18例)和非双LVIS支架组(NDLS组,27例),对比2组手术效果、围手术期并发症及随访结果。结果DLS组、NDLS组术后即刻完全栓塞率分别为72.22%(13/18)和55.56%(15/27),围手术期并发症发生率分别为16.67%(3/18)、25.93%(7/27),组间差异均无统计学意义(P均>0.05)。术后3、6个月,2组神经功能恢复良好率差异均无统计学意义(P均>0.05)。术后3个月DLS组复发率15.38%(2/13),低于NDLS组的57.89%(11/19,P=0.03);术后6个月2组复发率(0 vs 13.33%)差异无统计学意义(P>0.05)。结论双LVIS支架套叠辅助弹簧圈栓塞治疗BBA安全、有效,能降低术后3个月复发率。展开更多
文摘目的探究MicroRNA-124(miR-124)对脑缺血损伤小鼠神经功能的改善作用及其内在机制。方法①取32只C57小鼠随机分为假手术组和脑缺血组(n=16),TTC染色和神经行为学评分检测小鼠脑损伤后3天脑梗死面积和运动情况,RT-qPCR检测miR-124在两组间的表达变化。②培养原代神经元细胞,通过氧糖剥夺模拟缺血再灌注损伤,观察氧糖剥夺后0、72 h miR-124的变化趋势。③另取32只C57小鼠随机分为脑缺血组和脑缺血+miR-124组(n=16),TTC染色、转棒实验和肢体放置实验检测小鼠的运动功能改善情况。④细胞实验分为氧糖剥夺组和氧糖剥夺+miR-124组,通过免疫荧光实验检测两组神经元轴突的生长情况。Targetscan数据库预测miR-124下游靶基因,双荧光素酶基因报告和Western blot在原代神经元等上验证对照组和miR-124组下游靶基因第10号染色体缺失的磷酸酶及张力蛋白同源基因(the phosphate and tensin homolog deleted on chromosome ten,PTEN)的表达变化。上调PTEN,检测氧糖剥夺+miR-124和氧糖剥夺+miR-124+PTEN两组神经元轴突的生长情况。(5)在体实验中检测脑缺血+miR-124组和脑缺血+miR-124+Pten组(n=8)转棒实验和肢体放置情况。结果脑缺血再灌注后,小鼠脑梗死面积约达到40%(P<0.001),小鼠的神经认知功能显著降低(P<0.001),已达到重度认知功能损害;miR-124的表达较对照组明显升高(P<0.05);而进一步过表达miR-124能够促进氧糖剥夺后神经元轴突的生长(P<0.05),显著缩小脑梗死面积(P<0.05),且促进动物运动能力的恢复和改善(P<0.05);Targetscan数据库预测miR-124下游靶基因为PTEN,双荧光素酶基因报告和Western blot都证实miR-124能有效下调PTEN的表达(P<0.05);最后,上调PTEN逆转了miR-124促进神经元轴突的生长以及改善小鼠运动能力的效应(P<0.05)。结论miR-124通过下调PTEN促进脑缺血小鼠神经元轴突生长,且有助于小鼠运动功能的恢复。
文摘目的:探讨白藜芦醇(RES)预处理对氧糖剥夺(OGD)诱导的大鼠神经干细胞(NSCs)损伤的影响。方法:取孕15 d SD大鼠体内胎鼠脑组织原代培养NSCs,实验分为4组,分别为对照组、OGD模型组、RES+OGD处理组和RES+OGD+sirtinol处理组。利用CCK-8试剂盒检测各组NSCs的细胞活性,利用乳酸脱氢酶(LDH)检测各组NSCs的损伤程度。采用Western Blot检测各组NSCs中沉默信息调节因子1(SIRT1)和解偶联蛋白2(UCP2)的蛋白表达水平。结果:OGD损伤能够降低NSCs中SIRT1蛋白表达(P<0.05),RES预处理能够上调OGD损伤后NSCs中SIRT1蛋白的表达(P<0.05);RES预处理能够提高OGD损伤模型中NSCs的细胞活性(P<0.05),降低LDH漏出量(P<0.05),下调NSCs中UCP2的蛋白表达。SIRT1抑制剂sirtinol能够降低OGD损伤后NSCs的细胞活性(P<0.05),增加LDH漏出量(P<0.05),上调NSCs中UCP2的蛋白表达(P<0.05)。结论:RES预处理对OGD诱导的大鼠NSCs损伤具有保护作用,其作用机制可能与SIRT1-UCP2通路有关。
文摘Objective:To explore the effects of dopamine receptor D2(DRD2)on astrocytic dedifferentiation based on SOX2-regulated genes in neural stem cells(NSCs)and astrocytes.Methods:Immunofluorescence staining and SOX2-GFP mice were used to examine the lineage differentiation of SOX2-positive cells during the development of cerebral cortex.Primary NSCs/astrocytes culture,ChIP-seq and Western Blot were adopted to analyze and verify the expression of candidate genes.Pharmacological manipulation,neurosphere formation,photochemical ischemia,immunofluorescence staining and behavior tests were adopted to evaluate the effects of activating DRD2 signaling on astrocytic dedifferentiation.Results:Immunofluorescence staining demonstrated the NSC-astrocyte switch of SOX2-expression in the normal development of cerebral cortex.ChIP-seq revealed enrichment of DRD2 signaling by SOX2-bound enhancers in NSCs and SOX2-bound promoters in astrocytes.Western Blot and immunofluorescence staining verified the expression of DRD2 in NSCs and reactive astrocytes.Application of quinagolide hydrocholoride(QH),an agonist of DRD2,significantly promoted astrocytic dedifferentiation both in vitro and in vivo following ischemia.In addition,quinagolide hydrocholoride treatment improved locomotion recovery.Conclusion:Activating DRD2 signaling facilitates astrocytic dedifferentiation and may be used to treat ischemic stroke.
文摘目的观察双LVIS支架套叠辅助弹簧圈栓塞治疗颅内血泡样动脉瘤(BBA)的效果。方法回顾性分析45例接受支架辅助弹簧圈栓塞治疗的BBA患者资料,按治疗方式分为双LVIS支架组(DLS组,18例)和非双LVIS支架组(NDLS组,27例),对比2组手术效果、围手术期并发症及随访结果。结果DLS组、NDLS组术后即刻完全栓塞率分别为72.22%(13/18)和55.56%(15/27),围手术期并发症发生率分别为16.67%(3/18)、25.93%(7/27),组间差异均无统计学意义(P均>0.05)。术后3、6个月,2组神经功能恢复良好率差异均无统计学意义(P均>0.05)。术后3个月DLS组复发率15.38%(2/13),低于NDLS组的57.89%(11/19,P=0.03);术后6个月2组复发率(0 vs 13.33%)差异无统计学意义(P>0.05)。结论双LVIS支架套叠辅助弹簧圈栓塞治疗BBA安全、有效,能降低术后3个月复发率。
