中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2015年
5期
448-453
,共6页
谢东伟%赵信德%周忠保%黄柒金
謝東偉%趙信德%週忠保%黃柒金
사동위%조신덕%주충보%황칠금
脑出血%锰(Ⅲ)(四苯甲酸)卟啉%氧化-硝化应激
腦齣血%錳(Ⅲ)(四苯甲痠)卟啉%氧化-硝化應激
뇌출혈%맹(Ⅲ)(사분갑산)계람%양화-초화응격
Intracerebral hemorrhage%Mn(Ⅲ)tetrakis (4-benzoic acid) porphyrin%Oxidative and nitrative stress
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.
目的 探討金屬卟啉錳(Ⅲ)(四苯甲痠)卟啉(MnTBAP)對大鼠腦齣血的保護作用及其作用機製.方法 66隻大鼠按隨機數字錶法分為假手術組、對照組和實驗組,每組22隻.後2組大鼠應用立體定嚮註射自體尾動脈血法製作腦齣血模型,併在造模成功後30 min于實驗組大鼠患側腦室內註射2 μL MnTBAP(100 μg/μL),于對照組註射等體積生理鹽水.大鼠腦齣血後24 h採用免疫熒光染色檢測血腫週圍組織4-羥基壬烯醛(4-HNE)、3-硝基酪氨痠(3-NT)、8-羥基脫氧鳥苷(8-OHdG)、緊密連接蛋白ZO-1錶達情況及髓過氧化物酶(MPO)浸潤情況,Western blotting檢測ZO-1、基質金屬蛋白酶-9(MMP-9)蛋白錶達水平;大鼠腦齣血後24、72 h採用于濕重法檢測腦組織水含量,採用神經功能缺損評分進行行為學評分.結果 大鼠腦齣血後24h,與對照組比較,實驗組血腫週圍組織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)暘性細胞數及MMP-9蛋白錶達水平(0.75±0.07 vs 0.96±0.04)明顯降低,ZO-1蛋白錶達水平(0.74±0.05 vs 0.56±0.06)明顯增加,差異均有統計學意義(P＜0.05).大鼠腦齣血後24、72 h,與對照組比較,實驗組腦組織水含量(80.41％±0.69％vs 82.48％±0.94％;79.78％±0.65％vs 81.57％±0.82％)、神經功能缺損評分(9.33±1.37vs 11.33±1.51;6.17±0.98vs 9.50±1.38)明顯降低,差異均有統計學意義(P＜0.05).結論 大鼠腦齣血後早期應用MnTBAP能通過有效減輕氧化-硝化應激損傷,減少中性粒細胞浸潤,降低MMP-9錶達,減輕血腦屏障破壞,從而對腦齣血後腦損傷產生一定的保護作用.
목적 탐토금속계람맹(Ⅲ)(사분갑산)계람(MnTBAP)대대서뇌출혈적보호작용급기작용궤제.방법 66지대서안수궤수자표법분위가수술조、대조조화실험조,매조22지.후2조대서응용입체정향주사자체미동맥혈법제작뇌출혈모형,병재조모성공후30 min우실험조대서환측뇌실내주사2 μL MnTBAP(100 μg/μL),우대조조주사등체적생리염수.대서뇌출혈후24 h채용면역형광염색검측혈종주위조직4-간기임희철(4-HNE)、3-초기락안산(3-NT)、8-간기탈양조감(8-OHdG)、긴밀련접단백ZO-1표체정황급수과양화물매(MPO)침윤정황,Western blotting검측ZO-1、기질금속단백매-9(MMP-9)단백표체수평;대서뇌출혈후24、72 h채용우습중법검측뇌조직수함량,채용신경공능결손평분진행행위학평분.결과 대서뇌출혈후24h,여대조조비교,실험조혈종주위조직3-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)양성세포수급MMP-9단백표체수평(0.75±0.07 vs 0.96±0.04)명현강저,ZO-1단백표체수평(0.74±0.05 vs 0.56±0.06)명현증가,차이균유통계학의의(P＜0.05).대서뇌출혈후24、72 h,여대조조비교,실험조뇌조직수함량(80.41％±0.69％vs 82.48％±0.94％;79.78％±0.65％vs 81.57％±0.82％)、신경공능결손평분(9.33±1.37vs 11.33±1.51;6.17±0.98vs 9.50±1.38)명현강저,차이균유통계학의의(P＜0.05).결론 대서뇌출혈후조기응용MnTBAP능통과유효감경양화-초화응격손상,감소중성립세포침윤,강저MMP-9표체,감경혈뇌병장파배,종이대뇌출혈후뇌손상산생일정적보호작용.
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.
