胞红蛋白、HMGB1在血塞通干预大鼠心脏停搏后的表达机制

目的通过建立大鼠心脏停搏复苏模型和原代培养海马神经元模型,探讨在血塞通治疗大鼠心脏骤停后,胞红蛋白、高迁移率族蛋白B1(HMGB1)发挥的作用机制。方法将实验大鼠分为假手术(SO)组、心肺复苏(CPR)组和干预治疗(IVT)组。在大鼠心脏停搏/CPR后3、12、24、48和72 h分别采用免疫组织化学法、蛋白免疫印迹法及实时荧光定量(qRT)-PCR法检测海马CA1区胞红蛋白、HMGB1的平均光密度值及其蛋白和基因的表达,ELISA法检测外周血清中胞红蛋白、HMGB1的浓度。在细胞上,通过原代培养海马神经元并建立缺氧缺糖(OGD/R)模型。结果在5个时间点,SO组中胞红蛋白、HMGB1的血清浓度,海马CA1区胞红蛋白和HMGB1的mRNA表达,体外培养海马神经元中胞红蛋白、HMGB1蛋白表达及其基因表达比较差异均无统计学意义(P均> 0.05);CPR组与SO组比较,差异均有统计学意义(P均<0.01);干预治疗组的表达水平与CPR组比较,各时间点差异亦均有统计学意义(P均<0.01)。在48h观察点,与SO组或对照组(未经OGD/R治疗)比较,CPR组中胞红蛋白的阳...

Mechanism of CyGB and HMGB1 expression after Xuesaitong intervention in rats with cardiac arrest

Objective To investigate the mechanism of CyGB and HMGB1 expression after Xuesaitong intervention in rats with cardiac arrest by establishing rat models of cardiac arrest/resuscitation and primary cultured hippocampal neurons. Methods All rats were divided into the sham operation （SO）, cardiopulmonary resuscitation （CPR） and intervention therapy （IVT） groups. At 3, 12, 24, 48 and 72 h after cardiac arrest/CPR, the average optical density values and the expression levels of CyGB and HMGB1 protein and gene in the hippocampal CA1 area were detected by immunohistochemical staining, Western blot and qRT-PCR, respectively. The concentrations of CyGB and HMGB1 in the peripheral serum were determined by ELISA. At the cellular level, primary hippocampal neurons were cultured, and an oxygen and glucose deprivation/reperfusion （OGD/R） model was established. Results SABC, ELISA, Western blot and qRT-PCR demonstrated that the serum concentrations of CyGB and HMGB1, the expression levels of CyGB and HMGB1 mRNA in the hippocampal CA1 area, and the expression levels of CyGB and HMGB1 protein and gene in the cultured hippocampal neurons did not significantly differ at five time points in the SO group （all P > 0.05）. Statistical significance was noted between the CPR and SO groups （P < 0.01）. Compared with the CPR group, the expression level at each time point was significantly different in the IVT group （all P < 0.01）. Compared with the SO or control group, the positive expression of CyGB protein at 48 h was down-regulated, whereas that of HMGB1 protein was up-regulated in the CPR group. In the OGD/R group （after OGD/R treatment）, the apoptosis rate of hippocampal neurons was significantly increased. Compared with the CPR or OGD/R group, the positive expression of CyGB protein was up-regulated, whereas that of HMGB1 protein was down-regulated in the IVT group. The apoptosis rate of hippocampal neurons was significantly reduced in the OGD/R group. Conclusion The expression level of CyGB in the cells is up-regulated and that of HMGB1 is down-regulated after Xuesaitong intervention in the treatment of cardiac arrest in rat models, which suppresses multiple signaling pathways, thereby inhibiting the release of other inflammatory cytokines, exerting anti-inflammatory and anti-apoptotic effects and playing a role in brain protection.