目的 探讨7,8-二羟基黄酮(7,8-DHF)调节酪氨酸激酶B受体(TrkB)/脑源性神经营养因子(BDNF)信号通路对脑出血小鼠的神经保护作用.方法 C57BL/6小鼠通过注射细菌胶原酶VⅡ构建脑出血小鼠模型.将造模成功的小鼠随机分为模型组、对照组(5 mg·kg-1 7,8-DHF)、实验组(1 mg·kg-1 K252a)、联合组(5 mg·kg-1 7,8-DHF+1 mg·kg-1 K252a),以注射0.9％NaCl的小鼠作为假手术组,每组10只.治疗结束后,用Garcia法对小鼠进行神经功能评分,用干湿重法检测脑组织脑含水量,用伊文思蓝法检测血脑屏障的通透性,用TUNEL法检测神经元凋亡情况,用蛋白质印迹法检测脑组织中TrkB、p-TrkB、BDNF蛋白的表达水平.结果 对照组、实验组、联合组、模型组和假手术组的神经功能评分分别为(15.47±1.55)、(7.23±0.73)、(10.55±1.06)、(10.45±1.05)和(16.12±1.62)分,脑含水量分别为(62.88±2.19)％、(83.77±3.11)％、(72.71±2.59)％、(72.88±2.61)％和(59.64±2.06)％,伊文思蓝含量分别为(3.26±0.33)、(16.23±1.63)、(8.78±0.88)、(9.47±0.95)和(1.02±0.11)μg·g-1,神经元凋亡率分别为(9.82±0.99)％、(39.88±3.99)％、(22.15±2.24)％、(25.71±2.58)％和(6.46±0.65)％,p-TrkB/TrkB 比值分别为 1.01±0.11、0.21±0.03、0.48±0.05、0.49±0.05和1.03±0.11,BDNF蛋白相对表达水平分别为1.15±0.12、0.18±0.02、0.46±0.05、0.42±0.05 和 1.18±0.12.假手术组、对照组、实验组的上述指标与模型组比较,对照组、实验组的上述指标与联合组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 7,8-DHF对脑出血小鼠具有神经保护作用,其机制可能与激活TrkB/BDNF信号通路有关.
Objective To investigate the neuroprotective effect of 7,8-dihydroxyflavone(7,8-DHF)on cerebral hemorrhage in mice by regulating the tyrosine kinase receptor B(TrkB)/brain derived neurotrophic factor(BDNF)signal pathway.Methods C57BL/6 mice were injected with bacterial collagenase V Ⅱ to establish cerebral hemorrhage model.The mice were randomly grouped into model group,control group(5 mg·kg-1 7,8-DHF),experimental group(1 mg·kg-1 K252a),and combined group(5 mg·kg-1 7,8-DHF+1 mg·kg-1 K252a),mice injected with normal saline were used as sham-operation group,with 10 mice in each group.After the treatment,the mice were scored for neurological function by Garcia method,brain water contents of the brain tissue were detected by the dry and wet weight method,the blood brain barrier permeability was examined using the Evans blue method,the neuronal apoptosis was detected by TUNEL method,and the protein expression levels of TrkB,phosphorylated TrkB(p-TrkB)and BDNF were detected by Western blot.Results The neurological function scores of control,experimental,combined,model and sham-operation groups were(15.47±1.55),(7.23±0.73),(10.55±1.06),(10.45±1.05)and(16.12±1.62)points;the brain water contents were(62.88±2.19)％,(83.77±3.11)％,(72.71±2.59)％,(72.88±2.61)％and(59.64±2.06)％;the Evans blue contents were(3.26±0.36),(16.23±1.63),(8.78±0.88),(9.47±0.95)and(1.02±0.11)μg·g-1;neuronal apoptosis rates were(9.82±0.99)％,(39.88±3.99)％,(22.15±2.24)％,(25.71±2.58)％and(6.46±0.65)％;p-TrkB/TrkB ratios were 1.01±0.11,0.21±0.03,0.48±0.05,0.49±0.05 and 1.03±0.11;the protein expression levels of BDNF were 1.15±0.12,0.18±0.02,0.46±0.05,0.42±0.05 and 1.18±0.12,respectively.The above indexes of sham-operation,control and experimental groups were compared with those of model group,and the above indexes of control and experimental groups were compared with those of combined group,the differences were statistically significant(all P＜0.05).Conclusion 7,8-DHF has neuroprotective effect on mice with intracerebral hemorrhage,and its mechanism may be related to the activation of TrkB/BDNF signal pathway.
王坤;文丽波;付金霞;张贺男;韩丽君
齐齐哈尔医学院附属第二医院神经内科,黑龙江齐齐哈尔 161000齐齐哈尔医学院生理学教研室,黑龙江齐齐哈尔 161000齐齐哈尔医学院病理学教研室,黑龙江齐齐哈尔 161000
中医学
7,8-二羟基黄酮;脑出血;酪氨酸激酶B受体;脑源性神经营养因子;神经保护
7,8-dihydroxyflavone;cerebral hemorrhage;tyrosine kinase receptor B;brain derived neurotrophic factor;neuroprotection
《中国临床药理学杂志》 2024 (001)
52-56 / 5
黑龙江省省属本科高校基本科研业务费科学技术研究基金资助项目(2020-KYYWF-0027)
10.13699/j.cnki.1001-6821.2024.01.011
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