国际脑血管病杂志
國際腦血管病雜誌
국제뇌혈관병잡지
INTERNATIONAL JOURNAL OF CEREBROVASCULAR DISEASES
2013年
2期
108-113
,共6页
脑缺血%环氧水解酶类%酶抑制剂%细胞凋亡%原癌基因蛋白质c-akt%疾病模型,动物%大鼠
腦缺血%環氧水解酶類%酶抑製劑%細胞凋亡%原癌基因蛋白質c-akt%疾病模型,動物%大鼠
뇌결혈%배양수해매류%매억제제%세포조망%원암기인단백질c-akt%질병모형,동물%대서
Brain Ischemia%Epoxide Hydrolases%Enzyme Inhibitors%Apoptosis%Proto-Oncogene Proteins c-akt%Disease Models,Animal%Rats
目的 探讨可溶性环氧物酶(soluble epoxide hydrolase,sEH)抑制剂12-(3-金刚烷-1-基脲基)-十二烷酸[12-(3-adamant an-1-yl-ureido)-dodec-anoic acid,AUDA]对局灶性脑缺血再灌注大鼠的神经保护作用和机制.方法 60只雄性Sprague-Dawley大鼠随机分为假手术组、生理盐水对照组以及小剂量(0.157 ml/kg)、中剂量(0.235 ml/kg)和大剂量(0.314 ml/kg) AUDA处理组(每组12只),各组随机取4只大鼠分别用于梗死体积、细胞凋亡和p-Akt免疫组织化学检测.线栓法建立大脑中动脉闭塞再灌注模型.各AUDA处理组和生理盐水对照组均在再灌注前分别经腹腔给予相应剂量的AUDA或等体积生理盐水.再灌注24 h时进行神经功能缺损评分.2,3,5-氯化三苯基四氮唑染色法检测脑梗死体积.原位缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测梗死周围区脑组织细胞凋亡.免疫组织化学法检测梗死周围区脑组织p-Akt表达.结果 TTC染色显示,假手术组未见梗死.生理盐水对照组以及小剂量、中剂量和大剂量AUDA组梗死体积分别为(254.146±25.481)、(212.679±7.514)、(150.188±33.997)和(99.563±3.415) mm3,存在显著性差异(F=39.637,P=0.000).各剂量AUDA组均显著性小于对照组(P均=0.000).中剂量AUDA组显著性小于小剂量AUDA组(P=0.002),而大剂量AUDA也显著性小于小剂量AUDA组(P =0.000)和中剂量AUDA组(P=0.006).TUNEL染色法显示,假手术组仅可见少量凋亡细胞[(6.400±1.477)个/高倍视野].生理盐水对照组以及小剂量、中剂量和大剂量AUDA组凋亡细胞数量分别为(57.550±13.067)、(47.030±8.423)、(34.530±4.393)和(26.400±2.683)个/高倍视野,各剂量AUDA组显著性少于生理盐水对照组(P均＜0.01),中剂量和大剂量AUDA组显著性少于小剂量AUDA组(P均＜0.01),大剂量AUDA组也显著性少于中剂量AUDA组(P＜0.01).免疫组织化学显示,假手术组仅可见少量p-Akt阳性细胞[(3.325±1.438)个/高倍视野],生理盐水对照组以及小剂量、中剂量和大剂量AUDA组p-Akt阳性细胞数量分别为(9.450 ±2.531)、(16.400 ±3.865)、(22.875±7.974)和(29.300±3.203)个/高倍视野,各剂量AUDA组显著性多于生理盐水对照组(P均＜0.01),中剂量和大剂量AUDA组显著性多于小剂量AUDA组(P均＜0.01),大剂量AUDA组也显著性多于中剂量AUDA组(P＜0.01).结论 抑制sEH可能通过上调PI3K/Akt通路减少梗死周围区神经元凋亡和缩小梗死体积,对局灶性脑缺血再灌注大鼠具有神经保护作用.
目的 探討可溶性環氧物酶(soluble epoxide hydrolase,sEH)抑製劑12-(3-金剛烷-1-基脲基)-十二烷痠[12-(3-adamant an-1-yl-ureido)-dodec-anoic acid,AUDA]對跼竈性腦缺血再灌註大鼠的神經保護作用和機製.方法 60隻雄性Sprague-Dawley大鼠隨機分為假手術組、生理鹽水對照組以及小劑量(0.157 ml/kg)、中劑量(0.235 ml/kg)和大劑量(0.314 ml/kg) AUDA處理組(每組12隻),各組隨機取4隻大鼠分彆用于梗死體積、細胞凋亡和p-Akt免疫組織化學檢測.線栓法建立大腦中動脈閉塞再灌註模型.各AUDA處理組和生理鹽水對照組均在再灌註前分彆經腹腔給予相應劑量的AUDA或等體積生理鹽水.再灌註24 h時進行神經功能缺損評分.2,3,5-氯化三苯基四氮唑染色法檢測腦梗死體積.原位缺口末耑標記法(TdT-mediated dUTP nick end labeling,TUNEL)檢測梗死週圍區腦組織細胞凋亡.免疫組織化學法檢測梗死週圍區腦組織p-Akt錶達.結果 TTC染色顯示,假手術組未見梗死.生理鹽水對照組以及小劑量、中劑量和大劑量AUDA組梗死體積分彆為(254.146±25.481)、(212.679±7.514)、(150.188±33.997)和(99.563±3.415) mm3,存在顯著性差異(F=39.637,P=0.000).各劑量AUDA組均顯著性小于對照組(P均=0.000).中劑量AUDA組顯著性小于小劑量AUDA組(P=0.002),而大劑量AUDA也顯著性小于小劑量AUDA組(P =0.000)和中劑量AUDA組(P=0.006).TUNEL染色法顯示,假手術組僅可見少量凋亡細胞[(6.400±1.477)箇/高倍視野].生理鹽水對照組以及小劑量、中劑量和大劑量AUDA組凋亡細胞數量分彆為(57.550±13.067)、(47.030±8.423)、(34.530±4.393)和(26.400±2.683)箇/高倍視野,各劑量AUDA組顯著性少于生理鹽水對照組(P均＜0.01),中劑量和大劑量AUDA組顯著性少于小劑量AUDA組(P均＜0.01),大劑量AUDA組也顯著性少于中劑量AUDA組(P＜0.01).免疫組織化學顯示,假手術組僅可見少量p-Akt暘性細胞[(3.325±1.438)箇/高倍視野],生理鹽水對照組以及小劑量、中劑量和大劑量AUDA組p-Akt暘性細胞數量分彆為(9.450 ±2.531)、(16.400 ±3.865)、(22.875±7.974)和(29.300±3.203)箇/高倍視野,各劑量AUDA組顯著性多于生理鹽水對照組(P均＜0.01),中劑量和大劑量AUDA組顯著性多于小劑量AUDA組(P均＜0.01),大劑量AUDA組也顯著性多于中劑量AUDA組(P＜0.01).結論 抑製sEH可能通過上調PI3K/Akt通路減少梗死週圍區神經元凋亡和縮小梗死體積,對跼竈性腦缺血再灌註大鼠具有神經保護作用.
목적 탐토가용성배양물매(soluble epoxide hydrolase,sEH)억제제12-(3-금강완-1-기뇨기)-십이완산[12-(3-adamant an-1-yl-ureido)-dodec-anoic acid,AUDA]대국조성뇌결혈재관주대서적신경보호작용화궤제.방법 60지웅성Sprague-Dawley대서수궤분위가수술조、생리염수대조조이급소제량(0.157 ml/kg)、중제량(0.235 ml/kg)화대제량(0.314 ml/kg) AUDA처리조(매조12지),각조수궤취4지대서분별용우경사체적、세포조망화p-Akt면역조직화학검측.선전법건립대뇌중동맥폐새재관주모형.각AUDA처리조화생리염수대조조균재재관주전분별경복강급여상응제량적AUDA혹등체적생리염수.재관주24 h시진행신경공능결손평분.2,3,5-록화삼분기사담서염색법검측뇌경사체적.원위결구말단표기법(TdT-mediated dUTP nick end labeling,TUNEL)검측경사주위구뇌조직세포조망.면역조직화학법검측경사주위구뇌조직p-Akt표체.결과 TTC염색현시,가수술조미견경사.생리염수대조조이급소제량、중제량화대제량AUDA조경사체적분별위(254.146±25.481)、(212.679±7.514)、(150.188±33.997)화(99.563±3.415) mm3,존재현저성차이(F=39.637,P=0.000).각제량AUDA조균현저성소우대조조(P균=0.000).중제량AUDA조현저성소우소제량AUDA조(P=0.002),이대제량AUDA야현저성소우소제량AUDA조(P =0.000)화중제량AUDA조(P=0.006).TUNEL염색법현시,가수술조부가견소량조망세포[(6.400±1.477)개/고배시야].생리염수대조조이급소제량、중제량화대제량AUDA조조망세포수량분별위(57.550±13.067)、(47.030±8.423)、(34.530±4.393)화(26.400±2.683)개/고배시야,각제량AUDA조현저성소우생리염수대조조(P균＜0.01),중제량화대제량AUDA조현저성소우소제량AUDA조(P균＜0.01),대제량AUDA조야현저성소우중제량AUDA조(P＜0.01).면역조직화학현시,가수술조부가견소량p-Akt양성세포[(3.325±1.438)개/고배시야],생리염수대조조이급소제량、중제량화대제량AUDA조p-Akt양성세포수량분별위(9.450 ±2.531)、(16.400 ±3.865)、(22.875±7.974)화(29.300±3.203)개/고배시야,각제량AUDA조현저성다우생리염수대조조(P균＜0.01),중제량화대제량AUDA조현저성다우소제량AUDA조(P균＜0.01),대제량AUDA조야현저성다우중제량AUDA조(P＜0.01).결론 억제sEH가능통과상조PI3K/Akt통로감소경사주위구신경원조망화축소경사체적,대국조성뇌결혈재관주대서구유신경보호작용.
Objective To investigate the neuroprotective effect of soluble epoxide hydrolase (sEH) inhibitor 12-(3-adamantan-l-yl-ureido) dodecanoic acid (AUDA) on focal cerebral ischemia/reperfusion in rats and its mechanisms.Methods Sixty male Sprague-Dawley rats were randomly divided into sham operation and saline control groups,as well as low-dose (0.157 ml/kg),medium-dose (0.235 ml/kg) and high-dose (0.314 ml/kg) AUDA groups (n =12 in each group).Four rats in each group were selected for infarct volume,cell apoptosis and p-Akt immunohistochemistry detection.A model of middle cerebral artery ischemia/ reperfusion was induced by the suture method.The corresponding dose AUDA or equal volume of saline was injected intraperitoneally before reperfusion in each AUDA group and the saline control group.Neurological deficit scores were performed at 24 h of reperfusion.2,3,5 triphenyltetrazolium chloride (TTC) staining was used to detect infarct volume.TdT-mediated dUTP nick end labeling (TUNEL) was used to detect apoptotic cells of brain tissue in the periinfarction area.Immunohistochemical method was used to detect p-Akt expression of brain tissue in the peri-infarction area.Results TTC staining showed no infarction was observed in the sham operation group.The infarction volumes in the saline control group as well as the low-dose,medlum-dose and high-dose AUDA groups were 254.146 ± 25.481,212.679 ± 7.514,150.188 ± 33.997,and 99.563 ± 3.415 mm3,respectively.There were significant differences (F =39.637,P =0.000).The each dose AUDA group was significant less than the control group (all P=0.000).The medium-dose AUDA group was significantly less than the low-dose AUDA group (P=0.002),and the high-dose AUDA group was also significantly less than the low-dose AUDA group (P =0.000) and medium-dose AUDA group (P =0.006).TUNEL staining showed that a small number of apoptotic cells (6.400 ± 1.477/high-power field) were observed in the sham operation group.The numbers of apoptotic cells in the saline control group as well as in the low-dose,medium-dose and high-dose AUDA groups were 57.550 ± 13.067,47.030 ± 8.423,34.530 ± 4.393 and 26.400 ± 2.683/high power field,respectively.Each dose AUDA group was significantly less than the saline control group (all P ＜0.01).The medium-dose and high-dose AUDA groups were significantly less than the low-dose AUDA group (P ＜ 0.01),and the high-dose AUDA group was also significantly less than the medium-dose AUDA group (P ＜0.01).Immunohistochemistry showed that only a few p-Akt-positive cells (3.325 ± 1.438/high power field) were observed in the sham operation group.The numbers of p-Akt-positive cells in the saline control group as well as the low-dose,medium-dose and high-dose AUDA groups were 9.450 ±2.531,16.400 ± 3.865,22.875 ± 7.974,and 29.300 ± 3.203/high-power field,respectively.Each dose AUDA group was significantly more than the saline control group (all P ＜0.01).The medium-dose and high-dose AUDA groups were significantly more than the low-dose AUDA group (all P ＜0.01).The high-dose AUDA group was also significantly more than the medium-dose AUDA group (P ＜ 0.01).Conclusions The inhibition of sEH may decrease neuronal apoptosis and reduce infarct volume in the peri-infarction area by upregulating the PI3K/Akt pathway.It has a neuroprotective effect for focal cerebral ischemia/reperfusion in rats.
