中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2014年
8期
632-634
,共3页
苏云洁%李熙鸿%杨欣%王静%屈艺%母得志
囌雲潔%李熙鴻%楊訢%王靜%屈藝%母得誌
소운길%리희홍%양흔%왕정%굴예%모득지
脓毒症%脑损伤%皮质%凋亡%含半胱氨酸的天冬氨酸蛋白水解酶-3
膿毒癥%腦損傷%皮質%凋亡%含半胱氨痠的天鼕氨痠蛋白水解酶-3
농독증%뇌손상%피질%조망%함반광안산적천동안산단백수해매-3
Sepsis%Brain injury%Cortex%Apoptosis%Cysteinyl aspartate specific proteinase-3
目的 探讨脓毒症大鼠大脑皮质神经元含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)表达的意义.方法 行盲肠结扎穿孔术(CLP)建立脓毒症模型.70只30日龄健康雄性Wistar大鼠,按简单随机法分为CLP组(50只)和对照组(20只).CLP组于CLP手术后6h、12 h、24 h、48 h,分别取5只大鼠做神经行为学检查,然后处死,取脑组织,采用免疫荧光染色和Western blot法检测大鼠大脑皮质凋亡蛋白caspase-3的表达.对照组不给予CLP,其余处理同CLP组.结果 CLP手术后12 h、24 h和48 h大鼠的神经行为学评分均明显低于对照组(=3.651、3.773、7.155,P均＜0.05),且随着时间变化,大鼠的神经行为评分逐渐降低,大鼠的整体情况越来越差.免疫荧光检测表明:对照组大鼠大脑皮质中仅表达微量的caspase-3蛋白,而在CLP手术后12 h,脓毒症大鼠大脑皮质中caspase-3蛋白表达明显增加.Western blot检测表明:CLP手术后6h、12 h和24h大鼠大脑皮质caspase-3蛋白表达均明显高于对照组(P均＜0.05).CLP手术后6h,caspase-3蛋白表达开始升高,12 h达高峰,48 h下降.结论 脓毒症时大鼠的神经行为学评分降低,大脑皮质神经元凋亡蛋白酶caspase-3表达上调.
目的 探討膿毒癥大鼠大腦皮質神經元含半胱氨痠的天鼕氨痠蛋白水解酶-3(caspase-3)錶達的意義.方法 行盲腸結扎穿孔術(CLP)建立膿毒癥模型.70隻30日齡健康雄性Wistar大鼠,按簡單隨機法分為CLP組(50隻)和對照組(20隻).CLP組于CLP手術後6h、12 h、24 h、48 h,分彆取5隻大鼠做神經行為學檢查,然後處死,取腦組織,採用免疫熒光染色和Western blot法檢測大鼠大腦皮質凋亡蛋白caspase-3的錶達.對照組不給予CLP,其餘處理同CLP組.結果 CLP手術後12 h、24 h和48 h大鼠的神經行為學評分均明顯低于對照組(=3.651、3.773、7.155,P均＜0.05),且隨著時間變化,大鼠的神經行為評分逐漸降低,大鼠的整體情況越來越差.免疫熒光檢測錶明:對照組大鼠大腦皮質中僅錶達微量的caspase-3蛋白,而在CLP手術後12 h,膿毒癥大鼠大腦皮質中caspase-3蛋白錶達明顯增加.Western blot檢測錶明:CLP手術後6h、12 h和24h大鼠大腦皮質caspase-3蛋白錶達均明顯高于對照組(P均＜0.05).CLP手術後6h,caspase-3蛋白錶達開始升高,12 h達高峰,48 h下降.結論 膿毒癥時大鼠的神經行為學評分降低,大腦皮質神經元凋亡蛋白酶caspase-3錶達上調.
목적 탐토농독증대서대뇌피질신경원함반광안산적천동안산단백수해매-3(caspase-3)표체적의의.방법 행맹장결찰천공술(CLP)건립농독증모형.70지30일령건강웅성Wistar대서,안간단수궤법분위CLP조(50지)화대조조(20지).CLP조우CLP수술후6h、12 h、24 h、48 h,분별취5지대서주신경행위학검사,연후처사,취뇌조직,채용면역형광염색화Western blot법검측대서대뇌피질조망단백caspase-3적표체.대조조불급여CLP,기여처리동CLP조.결과 CLP수술후12 h、24 h화48 h대서적신경행위학평분균명현저우대조조(=3.651、3.773、7.155,P균＜0.05),차수착시간변화,대서적신경행위평분축점강저,대서적정체정황월래월차.면역형광검측표명:대조조대서대뇌피질중부표체미량적caspase-3단백,이재CLP수술후12 h,농독증대서대뇌피질중caspase-3단백표체명현증가.Western blot검측표명:CLP수술후6h、12 h화24h대서대뇌피질caspase-3단백표체균명현고우대조조(P균＜0.05).CLP수술후6h,caspase-3단백표체개시승고,12 h체고봉,48 h하강.결론 농독증시대서적신경행위학평분강저,대뇌피질신경원조망단백매caspase-3표체상조.
Objective To investigate the expression of apoptotic protease cysteinyl aspartate specific proteinase (caspase-3) in cortex neurons of rats with sepsis.Methods Models of rats with sepsis were established by the cecal ligation and puncture (CLP).Totally 70 cases of 30-day-old male Wistar rats were randomly divided into CLP group (n =50) and control group (n =20).In CLP group,CLP was performed in the rats.Neurobehavioral score was measured in 5 rats at 6,12,24 and 48 h after CLP surgery,respectively.Then,they were killed and their brains were removed.The immunohistochemical staining and Western blot were used to detect the apoptotic protein caspase-3 expression in cortex of rats.Control group did not undergo CLP,and the other treatment was the same as CLP group.Results Neurobehavioral scores at 12,24 and 48 h after CLP surgery were significantly lower than that in the control group(t =3.651,3.773,7.155,all P ＜ 0.05),and the scores were gradually decreased,overall situation of rats was getting worse along with the time.Caspase-3 protein expressed only in trace amounts in rat cerebral cortex in the control group by immunofluorescence analysis,however,its expression was significantly increased at 12 h after CLP surgery.Western blot test showed that caspase-3 protein expression in rat cerebral cortex at 6,12 and 24 h after CLP surgery was significantly higher than those in control group (all P ＜ 0.05).Its expression began to increase at 6 h after CLP surgery,and reached the peak at 12 h,then decreased at 48 h.Conclusion The neurobehavioral scores decreases and the expression of apeptosis protease caspase-3 increases in cortex of rats with sepsis brain injury.
