目的:探究半乳糖凝集素-3 (Galectin-3)对大鼠脑出血后脑损伤的影响及相关机制。方法:将120只清洁级Wistar大鼠随机分为假手术组(Sham组)、脑出血模型组(ICH组)、Galectin-3抑制剂MCP组(MCP组),每组40只。采用胶原酶脑内定点注射法制作...目的:探究半乳糖凝集素-3 (Galectin-3)对大鼠脑出血后脑损伤的影响及相关机制。方法:将120只清洁级Wistar大鼠随机分为假手术组(Sham组)、脑出血模型组(ICH组)、Galectin-3抑制剂MCP组(MCP组),每组40只。采用胶原酶脑内定点注射法制作大鼠ICH模型,Sham组和ICH组大鼠经侧脑室注射20 μg正常绵羊IgG,MCP组大鼠注射4 μg MCP。48 h后采用Longa评分标准进行神经功能缺损评分,并检测各组大鼠脑含水量,评估血脑屏障通透性,TUNEL染色法检测神经细胞凋亡,酶联免疫吸附法(ELISA)检测白细胞介素-1β (IL-1β)、白细胞介素-6 (IL-6)和肿瘤坏死因子-α (TNF-α)含量,蛋白免疫印迹法(Western Blot)检测TLR-4/NF-κB通路相关蛋白表达。结果:与Sham组相比,ICH组大鼠神经功能缺损评分、脑含水量、血脑屏障通透性、细胞凋亡率、脑组织IL-1β、IL-6、TNF-α含量升高(P κB p65蛋白表达量增加(P β、IL-6、TNF-α含量降低(P κB p65蛋白表达量降低(P κB通路有关。Objective: To explore the effect of Galectin-3 on brain injury after cerebral hemorrhage in rats and its related mechanism. Methods: A total of 120 Wistar rats were randomly divided into Sham group, ICH group, Galectin-3 inhibitor MCP group (MCP group), 40 mice per group. ICH model was made by intracerebral injection of collagenase. 20 μg normal sheep IgG was injected into the lateral ventricle of Sham group and ICH group, and 4 μg MCP was injected into the MCP group. 48 h later, Longa score was used to evaluate the neural function deficit, and the brain water content of rats in each group was detected to evaluate the blood-brain barrier permeability. TUNEL staining was used to detect nerve cell apoptosis. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA), and the expression of TLR-4/NF-κB pathway related proteins was detected by Western Blot. Results: Compared with Sham group, the neural function deficit score, brain water content, blood-brain barrier permeability, cell apoptosis rate, the contents of IL-1β, IL-6 and TNF-α in brain tissue were increased in ICH group (P κB p65 in brain tissue were increased (P β, IL-6 and TNF-α in brain tissue were decreased in MCP group (P κB p65 in brain tissue were decreased (P κB pathway.展开更多
为检测鸡抗病毒相关基因在病毒感染过程中的动态变化规律,研究建立了鸡IFN-a、IFN-β、TLR-3和TLR-7的实时荧光定量RT-PCR的检测方法。根据GenBank提供的鸡IFN-α、IFN-β、FLR-3和TLR-7参考序列设计了相应的特异引物,用常规RT-PCR方法...为检测鸡抗病毒相关基因在病毒感染过程中的动态变化规律,研究建立了鸡IFN-a、IFN-β、TLR-3和TLR-7的实时荧光定量RT-PCR的检测方法。根据GenBank提供的鸡IFN-α、IFN-β、FLR-3和TLR-7参考序列设计了相应的特异引物,用常规RT-PCR方法从鸡脾脏总RNA中扩增得到鸡相应基因。将其cDNA分别克隆到pMD19-T载体中进行测序鉴定,采用SYBR Green I染料法,建立标准曲线并分析其溶解曲线。结果表明,这4种基因的检测灵敏度可高达46 copies/μL,且具有良好的特异性和重复性。此检测方法的建立为动态定量检测及研究鸡IFN-α、IFN-β、TLR-3和TLR-7基因的表达变化提供了有效的手段。展开更多
文摘目的:探究半乳糖凝集素-3 (Galectin-3)对大鼠脑出血后脑损伤的影响及相关机制。方法:将120只清洁级Wistar大鼠随机分为假手术组(Sham组)、脑出血模型组(ICH组)、Galectin-3抑制剂MCP组(MCP组),每组40只。采用胶原酶脑内定点注射法制作大鼠ICH模型,Sham组和ICH组大鼠经侧脑室注射20 μg正常绵羊IgG,MCP组大鼠注射4 μg MCP。48 h后采用Longa评分标准进行神经功能缺损评分,并检测各组大鼠脑含水量,评估血脑屏障通透性,TUNEL染色法检测神经细胞凋亡,酶联免疫吸附法(ELISA)检测白细胞介素-1β (IL-1β)、白细胞介素-6 (IL-6)和肿瘤坏死因子-α (TNF-α)含量,蛋白免疫印迹法(Western Blot)检测TLR-4/NF-κB通路相关蛋白表达。结果:与Sham组相比,ICH组大鼠神经功能缺损评分、脑含水量、血脑屏障通透性、细胞凋亡率、脑组织IL-1β、IL-6、TNF-α含量升高(P κB p65蛋白表达量增加(P β、IL-6、TNF-α含量降低(P κB p65蛋白表达量降低(P κB通路有关。Objective: To explore the effect of Galectin-3 on brain injury after cerebral hemorrhage in rats and its related mechanism. Methods: A total of 120 Wistar rats were randomly divided into Sham group, ICH group, Galectin-3 inhibitor MCP group (MCP group), 40 mice per group. ICH model was made by intracerebral injection of collagenase. 20 μg normal sheep IgG was injected into the lateral ventricle of Sham group and ICH group, and 4 μg MCP was injected into the MCP group. 48 h later, Longa score was used to evaluate the neural function deficit, and the brain water content of rats in each group was detected to evaluate the blood-brain barrier permeability. TUNEL staining was used to detect nerve cell apoptosis. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA), and the expression of TLR-4/NF-κB pathway related proteins was detected by Western Blot. Results: Compared with Sham group, the neural function deficit score, brain water content, blood-brain barrier permeability, cell apoptosis rate, the contents of IL-1β, IL-6 and TNF-α in brain tissue were increased in ICH group (P κB p65 in brain tissue were increased (P β, IL-6 and TNF-α in brain tissue were decreased in MCP group (P κB p65 in brain tissue were decreased (P κB pathway.
文摘为检测鸡抗病毒相关基因在病毒感染过程中的动态变化规律,研究建立了鸡IFN-a、IFN-β、TLR-3和TLR-7的实时荧光定量RT-PCR的检测方法。根据GenBank提供的鸡IFN-α、IFN-β、FLR-3和TLR-7参考序列设计了相应的特异引物,用常规RT-PCR方法从鸡脾脏总RNA中扩增得到鸡相应基因。将其cDNA分别克隆到pMD19-T载体中进行测序鉴定,采用SYBR Green I染料法,建立标准曲线并分析其溶解曲线。结果表明,这4种基因的检测灵敏度可高达46 copies/μL,且具有良好的特异性和重复性。此检测方法的建立为动态定量检测及研究鸡IFN-α、IFN-β、TLR-3和TLR-7基因的表达变化提供了有效的手段。
