中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2013年
4期
373-378
,共6页
康健%龚平%任延波%高冬娜%丁琼蕾
康健%龔平%任延波%高鼕娜%丁瓊蕾
강건%공평%임연파%고동나%정경뢰
心肺复苏%低氧诱导因子-1α%促红细胞生成素%血管内皮生长因子%神经元特异性烯醇化酶%S100β蛋白%β-七叶皂苷钠%脑保护
心肺複囌%低氧誘導因子-1α%促紅細胞生成素%血管內皮生長因子%神經元特異性烯醇化酶%S100β蛋白%β-七葉皂苷鈉%腦保護
심폐복소%저양유도인자-1α%촉홍세포생성소%혈관내피생장인자%신경원특이성희순화매%S100β단백%β-칠협조감납%뇌보호
Cardiopulmonary resuscitation%Hypoxia-inducible factor-1 α%Erythropoietin%Vascular endothelial growth factor%Neuron specific enolase%S100β%β-sodium aescinate%Neuroprotection
目的 探讨自主循环恢复(return of spontaneous circulation,ROSC)后大鼠脑细胞中低氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)的表达及β-七叶皂苷钠对其影响.方法 清洁级SD成年大鼠60只,随机(随机数字法)分3组(n=20).(1)实验组:ROSC后即刻腹腔注射β-七叶皂苷钠(5 mg/kg)一次；(2)对照组:ROSC后即刻腹腔注射等量生理盐水一次；(3)假手术组:未经历心搏骤停-心肺复苏和未给药.采用窒息合并冰氯化钾停跳液法制备大鼠心搏骤停-心肺复苏模型.分别在ROSC后1、6、12、24 h抽血,然后处死取脑组织.ELISA检测血清NSE、S100β,免疫组化和Real-time PCR检测脑细胞HIF-1α、VEGF、EPO蛋白和mRNA.各组同一时间点之间比较采用成组t检验,组内不同时间点之间比较采用单因素方差分析,多重比较采用Bonferroni法.相关性分析采用Pearson法.结果 与假手术组比较,ROSC后1、6、12和24 h对照组大鼠血清NSE与S100β明显升高(P＜0.05),脑细胞中HIF-1 α、VEGF、EPO蛋白和mRNA表达均明显增加(P＜0.05).与对照组比较,ROSC后1、6、12、24 h实验组大鼠血清NSE与S100β明显降低(P＜0.05),脑细胞中HIF-1α、VEGF、EPO蛋白和mRNA表达均明显增加(P＜0.05).ROSC后大鼠脑细胞HIF-1αmRNA与EPO mRNA、VEGF mRNA均呈正相关(r=0.866,P＜0.05；r =0.952,P＜0.01).结论 心肺复苏后大鼠脑细胞HIF-1α表达增加,β-七叶皂苷钠能上调HIF-1α转录和蛋白表达,可能是通过增加EPO、VEGF基因转录及蛋白表达而增加脑细胞对缺血缺氧的耐受性,从而起神经保护作用.
目的 探討自主循環恢複(return of spontaneous circulation,ROSC)後大鼠腦細胞中低氧誘導因子-1α(hypoxia-inducible factor-1α,HIF-1α)的錶達及β-七葉皂苷鈉對其影響.方法 清潔級SD成年大鼠60隻,隨機(隨機數字法)分3組(n=20).(1)實驗組:ROSC後即刻腹腔註射β-七葉皂苷鈉(5 mg/kg)一次；(2)對照組:ROSC後即刻腹腔註射等量生理鹽水一次；(3)假手術組:未經歷心搏驟停-心肺複囌和未給藥.採用窒息閤併冰氯化鉀停跳液法製備大鼠心搏驟停-心肺複囌模型.分彆在ROSC後1、6、12、24 h抽血,然後處死取腦組織.ELISA檢測血清NSE、S100β,免疫組化和Real-time PCR檢測腦細胞HIF-1α、VEGF、EPO蛋白和mRNA.各組同一時間點之間比較採用成組t檢驗,組內不同時間點之間比較採用單因素方差分析,多重比較採用Bonferroni法.相關性分析採用Pearson法.結果 與假手術組比較,ROSC後1、6、12和24 h對照組大鼠血清NSE與S100β明顯升高(P＜0.05),腦細胞中HIF-1 α、VEGF、EPO蛋白和mRNA錶達均明顯增加(P＜0.05).與對照組比較,ROSC後1、6、12、24 h實驗組大鼠血清NSE與S100β明顯降低(P＜0.05),腦細胞中HIF-1α、VEGF、EPO蛋白和mRNA錶達均明顯增加(P＜0.05).ROSC後大鼠腦細胞HIF-1αmRNA與EPO mRNA、VEGF mRNA均呈正相關(r=0.866,P＜0.05；r =0.952,P＜0.01).結論 心肺複囌後大鼠腦細胞HIF-1α錶達增加,β-七葉皂苷鈉能上調HIF-1α轉錄和蛋白錶達,可能是通過增加EPO、VEGF基因轉錄及蛋白錶達而增加腦細胞對缺血缺氧的耐受性,從而起神經保護作用.
목적 탐토자주순배회복(return of spontaneous circulation,ROSC)후대서뇌세포중저양유도인자-1α(hypoxia-inducible factor-1α,HIF-1α)적표체급β-칠협조감납대기영향.방법 청길급SD성년대서60지,수궤(수궤수자법)분3조(n=20).(1)실험조:ROSC후즉각복강주사β-칠협조감납(5 mg/kg)일차；(2)대조조:ROSC후즉각복강주사등량생리염수일차；(3)가수술조:미경력심박취정-심폐복소화미급약.채용질식합병빙록화갑정도액법제비대서심박취정-심폐복소모형.분별재ROSC후1、6、12、24 h추혈,연후처사취뇌조직.ELISA검측혈청NSE、S100β,면역조화화Real-time PCR검측뇌세포HIF-1α、VEGF、EPO단백화mRNA.각조동일시간점지간비교채용성조t검험,조내불동시간점지간비교채용단인소방차분석,다중비교채용Bonferroni법.상관성분석채용Pearson법.결과 여가수술조비교,ROSC후1、6、12화24 h대조조대서혈청NSE여S100β명현승고(P＜0.05),뇌세포중HIF-1 α、VEGF、EPO단백화mRNA표체균명현증가(P＜0.05).여대조조비교,ROSC후1、6、12、24 h실험조대서혈청NSE여S100β명현강저(P＜0.05),뇌세포중HIF-1α、VEGF、EPO단백화mRNA표체균명현증가(P＜0.05).ROSC후대서뇌세포HIF-1αmRNA여EPO mRNA、VEGF mRNA균정정상관(r=0.866,P＜0.05；r =0.952,P＜0.01).결론 심폐복소후대서뇌세포HIF-1α표체증가,β-칠협조감납능상조HIF-1α전록화단백표체,가능시통과증가EPO、VEGF기인전록급단백표체이증가뇌세포대결혈결양적내수성,종이기신경보호작용.
Objective To investigate the expression of the hypoxia-inducible factor (HIF)-1α in rat brain neurons and the intervention of β-sodium aescinate after restoration of spontaneous circulation (ROSC).Methods Sixty SD adult rats were randomly (random number) divided into 3 groups (n =20),namely experiment group,control group and sham operation group.(1) The rats of experiment group were injected intraperitoneally with β-sodium aescinate (5 mg/kg) immediately after ROSC.(2) The rats of control group received normal saline injected intraperitoneally instead of β-sodium aescinate solution.(3)The rats of sham operation group did not have cardiac arrest and β-sodium aescinate intervention.Cardiac arrest rat model was established by using asphyxiation and intra-venous potassium chloride solution.Blood samples were taken 1 h,6 h,12 h and 24 h after ROSC,and subsequently rats were sacrificed and their brain tissues were harvested.The expressions of HIF-1 α mRNA,vascular endothelial growth factor (VEGF)mRNA and erythropoitin (EPO) mRNA and their protein levels in rat brain neurons were detected by using RT-PCR and immunohistochemistry,and the levels of serum neuron-specific enolase (NSE) and S100β proteins were determined by using enzyme-linked immunosorbent assay.The t test or one-way ANOVA was used to assess overall differences among groups for each of the variables,followed by Bonferroni test for multiple comparisons.Pearson method was used for correlation analysis.Results Compared with the sham operation group at intervals of 1 h,6 h,12 h and 24 h after ROSC,levels of serum S100β and NSE proteins were significantly increased in rats of the control group (P ＜ 0.05).Meanwhile,the expressions of HIF-1 α mRNA,VEGF mRNA and EPO mRNA and their protein levels in rat brain neurons were significantly increased in the control rats (P ＜0.05).Compared with the control group at intervals of 1 h,6 h,12 h and 24 h after ROSC,levels of serum NSE and S100β proteins were significantly decreased in rats of the experiment group (P ＜ 0.05).Whereas,the expressions of HIF-1 α mRNA,VEGF mRNA and EPO mRNA and their protein levels in rat brain neurons were significantly increased in rats of the experiment group (P ＜0.05).HIF-1 α mRNA was positively correlated with EPO mRNA and VEGF mRNAs (r =O.866,P ＜0.05 ; r =0.952,P ＜ O.01).Conclusions The expression of hypoxia-inducible factor-1 α is increased in rat brain cells after ROSC,and β-sodium aescinate up-regulates the expression of hypoxia-inducible factor1 α mRNA and protein levels.The up-regulated expression of hypoxia-inducible factor-1α improves the resistance of brain cells to ischemia and hypoxia contributing neuronal protection,which might be due to upregulated EPO and VEGF expressions induced by hypoxia-inducible factor-1α.
