β—七叶皂苷钠对心肺复苏后大鼠脑细胞中低氧诱导因子—1α的影响(1)
【摘要】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-1α、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-1α mRNA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。
【关键词】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
Effect of β-sodium aescinate on hypoxia-inducible factor-1α expression in rat brain neurons after cardiopulmonary resuscitation KANG Jian，GONG Ping，REN Yan-bo，GAO Dong-na，DING Qiong-lei.Department of Emergency，the First Hospital Affiliated to Dalian Medical University， Dalian 116011，China
Corresponding author： KANG Jian， Email： kangjian1998@yahoo.com.cn
【Abstract】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-1α 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-1α 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-1α 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-1α mRNA was positively correlated with EPO mRNA and VEGF mRNAs (r=0.866， P<0.05； r=0.952， P<0.01).Conclusions The expression of hypoxia-inducible factor-1α is increased in rat brain cells after ROSC， and β-sodium aescinate up-regulates the expression of hypoxia-inducible factor-1α 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 up-regulated EPO and VEGF expressions induced by hypoxia-inducible factor-1α.
, http://www.100md.com(康健 龚平 任延波 高冬娜 丁琼蕾)
【关键词】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
Effect of β-sodium aescinate on hypoxia-inducible factor-1α expression in rat brain neurons after cardiopulmonary resuscitation KANG Jian，GONG Ping，REN Yan-bo，GAO Dong-na，DING Qiong-lei.Department of Emergency，the First Hospital Affiliated to Dalian Medical University， Dalian 116011，China
Corresponding author： KANG Jian， Email： kangjian1998@yahoo.com.cn
【Abstract】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-1α 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-1α 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-1α 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-1α mRNA was positively correlated with EPO mRNA and VEGF mRNAs (r=0.866， P<0.05； r=0.952， P<0.01).Conclusions The expression of hypoxia-inducible factor-1α is increased in rat brain cells after ROSC， and β-sodium aescinate up-regulates the expression of hypoxia-inducible factor-1α 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 up-regulated EPO and VEGF expressions induced by hypoxia-inducible factor-1α.
, http://www.100md.com(康健 龚平 任延波 高冬娜 丁琼蕾)