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
目的观察结核分枝杆菌早期分泌靶抗原(ESAT-6)对γδT细胞产生淋巴因子的影响,并对其信号转导机制进行初步探讨。方法使用Ficoll密度梯度离心法分离健康人外周血单个核细胞(PBMC),用流式细胞术分选纯化γδT细胞;Toll样受体-4(TLR-4)特...目的观察结核分枝杆菌早期分泌靶抗原(ESAT-6)对γδT细胞产生淋巴因子的影响,并对其信号转导机制进行初步探讨。方法使用Ficoll密度梯度离心法分离健康人外周血单个核细胞(PBMC),用流式细胞术分选纯化γδT细胞;Toll样受体-4(TLR-4)特异性抑制剂E5564处理γδT细胞以抑制TLR-4的功能,并用蛋白质免疫印迹法(Western blot)及逆转录-PCR(RT-PCR)法检测γδT细胞TLR-4蛋白及mRNA的表达,以不加任何刺激因子的γδT细胞作空白对照,用(ESAT-6)刺激γδT细胞,并在第0、1、3、6、9和12d取细胞上清液,用酶联免疫吸附试验(ELISA)试剂盒检测IL-17、肿瘤坏死因子-α(TNF-α)及干扰素-γ(IFN-γ)的分泌水平。结果 Western blot及PCR结果显示ESAT-6能显著提高γδT细胞TLR-4基因的转录与蛋白的表达(P<0.01);ESAT-6能显著提高γδT细胞IL-17、TNF-α、IFN-γ的表达(P<0.01),而TLR-4特异性抑制剂E5564能显著的抑制ESAT-6所致的γδT细胞IL-17、TNF-α、IFN-γ淋巴因子的增加(P<0.01)。结论 ESAT-6能显著上调γδT细胞IL-17、TNF-α及IFN-γ的表达,其增强γδT细胞IL-17、TNF-α及IFN-γ表达的信号转导机制可能与TLR-4信号通路有关。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.
文摘目的观察结核分枝杆菌早期分泌靶抗原(ESAT-6)对γδT细胞产生淋巴因子的影响,并对其信号转导机制进行初步探讨。方法使用Ficoll密度梯度离心法分离健康人外周血单个核细胞(PBMC),用流式细胞术分选纯化γδT细胞;Toll样受体-4(TLR-4)特异性抑制剂E5564处理γδT细胞以抑制TLR-4的功能,并用蛋白质免疫印迹法(Western blot)及逆转录-PCR(RT-PCR)法检测γδT细胞TLR-4蛋白及mRNA的表达,以不加任何刺激因子的γδT细胞作空白对照,用(ESAT-6)刺激γδT细胞,并在第0、1、3、6、9和12d取细胞上清液,用酶联免疫吸附试验(ELISA)试剂盒检测IL-17、肿瘤坏死因子-α(TNF-α)及干扰素-γ(IFN-γ)的分泌水平。结果 Western blot及PCR结果显示ESAT-6能显著提高γδT细胞TLR-4基因的转录与蛋白的表达(P<0.01);ESAT-6能显著提高γδT细胞IL-17、TNF-α、IFN-γ的表达(P<0.01),而TLR-4特异性抑制剂E5564能显著的抑制ESAT-6所致的γδT细胞IL-17、TNF-α、IFN-γ淋巴因子的增加(P<0.01)。结论 ESAT-6能显著上调γδT细胞IL-17、TNF-α及IFN-γ的表达,其增强γδT细胞IL-17、TNF-α及IFN-γ表达的信号转导机制可能与TLR-4信号通路有关。
