摘要
目的 探讨金属卟啉锰（Ⅲ）（四苯甲酸）卟啉（MnTBAP）对大鼠脑出血的保护作用及其作用机制.方法 66只大鼠按随机数字表法分为假手术组、对照组和实验组,每组22只.后2组大鼠应用立体定向注射自体尾动脉血法制作脑出血模型,并在造模成功后30 min于实验组大鼠患侧脑室内注射2 μL MnTBAP（100 μg/μL）,于对照组注射等体积生理盐水.大鼠脑出血后24 h采用免疫荧光染色检测血肿周围组织4-羟基壬烯醛（4-HNE）、3-硝基酪氨酸（3-NT）、8-羟基脱氧鸟苷（8-OHdG）、紧密连接蛋白ZO-1表达情况及髓过氧化物酶（MPO）浸润情况,Western blotting检测ZO-1、基质金属蛋白酶-9（MMP-9）蛋白表达水平;大鼠脑出血后24、72 h采用于湿重法检测脑组织水含量,采用神经功能缺损评分进行行为学评分.结果 大鼠脑出血后24h,与对照组比较,实验组血肿周围组织3-NT （264.53±83.99vs 413.22±89.16）、4-HNE （245.64±73.10 vs 391.41±51.43）、8-OHdG（221.53±68.25vs 332.32±94.93）、MPO（296.14±66.34vs 431.59±102.68）阳性细胞数及MMP-9蛋白表达水平（0.75±0.07 vs 0.96±0.04）明显降低,ZO-1蛋白表达水平（0.74±0.05 vs 0.56±0.06）明显增加,差异均有统计学意义（P＜0.05）.大鼠脑出血后24、72 h,与对照组比较,实验组脑组织水含量（80.41％±0.69％vs 82.48％±0.94％;79.78％±0.65％vs 81.57％±0.82％）、神经功能缺损评分（9.33±1.37vs 11.33±1.51;6.17±0.98vs 9.50±1.38）明显降低,差异均有统计学意义（P＜0.05）.结论 大鼠脑出血后早期应用MnTBAP能通过有效减轻氧化-硝化应激损伤,减少中性粒细胞浸润,降低MMP-9表达,减轻血脑屏障破坏,从而对脑出血后脑损伤产生一定的保护作用.
Objective To explore the protective effect of Mn （Ⅲl）tetrakis （4-benzoic acid） porphyrin （MnTBAP） on rats after intracerebral hemorrhage （ICH） and its mechanism.Methods Sixty-six adult SD rats were randomized into sham-operated group,control group and experimental group （n=22）.Rats in the latter two groups were performed stereotactic injection of autologous tail arterial blood to induce ICH models;the rats in the experimental group were given 2 μL MnTBAP （100 μg/μL） by intracerebroventricular injection 30 min after ICH,while the rats in the control group were given normal saline of same volume.The expressions of 4-hydroxynonenonal （4-HNE,a marker of lipid peroxidation）,3-nitrotyrosine （3-NT,a reliable marker of protein nitration）,8-hydroxy-2＇-deoxyguanosine （8-OHdG,a marker of DNA oxidative damage）,Zonula occludens-1 （ZO-1,a kind of tight junction protein） and myeloperoxidase （MPO,a marker of neutrophil） in the perihematomal brain tissues 24 h after ICH were detected by immunofluorescence;protein expressions of ZO-1 and matrix metalloproteinase-9 （MMP-9） were detected by Western blotting 24 h after ICH;brain water content and modified neurological severity （mNSS） scores were measured 24 and 72 h after ICH.Results As compared with those in the control group,3-NT （264.53±83.99vs413.22±89.16）,4-HNE （245.64±73.10vs 391.41±51.43）,8-OHdG （221.53±68.25 vs 332.32±94.93）,MPO （296.14±66.34 vs 431.59±102.68） and MMP-9 （0.75±0.07 vs 0.96±0.04） expressions in perihematomal brains of experimental group were significantly decreased,while the expressions of ZO-1 （0.74±0.05 vs 0.56±0.06） were significantly increased （P〈0.05）.The mNSS scores （9.33±1.37 vs 11.33±1.51;6.17±0.98 vs 9.50±1.38） and brain water contents in the experimental group were significantly lower as compared with those in the control group 24 and 72 h after ICH （80.41%±0.69% vs 82.48%±0.94%;79.78%±0.65% vs 81.57%±0.82%） （P〈0.05）.Conclusion MnTBAP could protect injured brain tissues by alleviating oxidative and nitrative stress,decreasing neutrophils invasion and MMP-9 activation at early stage of ICH;meanwhile,MnTBAP could relieve the blood-brain barrier disruption and neurological deficit following ICH.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2015年第5期448-453,共6页
Chinese Journal of Neuromedicine
关键词
脑出血
锰(Ⅲ)(四苯甲酸)卟啉
氧化-硝化应激
Intracerebral hemorrhage
Mn（Ⅲ）tetrakis （4-benzoic acid） porphyrin
Oxidative and nitrative stress
