二氮嗪预处理对缺氧复氧后大鼠海马神经元凋亡的影响

目的研究线粒体内膜ATP敏感性钾通道（Mito-KAw）特异性开放剂二氮嗪预处理对缺氧复氧后大鼠海马神经元凋亡的影响。方法取离体培养的大鼠海马神经元，随机分为4组：对照组（A组）、二氮嗪30μmol／L组（B组）、二氮嗪1（30μmol／L组（C组）、二氮嗪100μmol／L＋Mito-KATP特异性阻断剂5．羟葵酸100μmol／L组（D组），各组神经元每天给予相应药物预处理1h，连续3d，继而缺氧4h复氧24h，观察神经元的活力、凋亡率、Bax和Bd．2蛋白的表达水平。结果与其它3组比较，C组海马神经元活力增强，凋亡率降低，Bcl．2蛋白表达水平升高，Bax蛋白表达水平下降（P〈0．01）。结论100μmol／L二氮嗪预处理通过改善Bcl-2与Bax蛋白表达的失衡，降低神经元的凋亡，对大鼠海马缺氧复氧神经元产生了保护效应。     

含二氮嗪停搏液对缺血再灌注心肌能量代谢保护作用研究

目的观察线粒体内膜三磷酸腺苷敏感钾离子通道开放剂二氮嗪加入停搏液内对缺血再灌注大鼠心肌能量代谢的保护作用.方法 50只大鼠随机分为五组,每组10只,取离体心脏;A组取正常心肌,其余制备离体工作心脏灌注模型;B组停灌30 min,复灌60 min;其余三组分别灌注不同的停搏液,C组灌注停搏液,D组灌注含二氮嗪的停搏液,E组灌注含二氮嗪停搏液之前10 min给予格列苯脲,各组复灌30 min后进行工作模式心脏灌注,监测心率、左室收缩峰压和舒张末压.实验终了取心肌,提取线粒体,测定细胞色素氧化酶活性、线粒体H -三磷酸腺苷酶活性;高效液相法测定心肌含量.结果缺血再灌注心肌线粒体细胞色素氧化酶活性和线粒体H -三磷酸腺苷酶活性降低(P＜0.05);而二氮嗪处理组相应指标显著改善(P＜0.01).结论含二氮嗪停搏液能够改善缺血再灌注心肌功能,能量代谢恢复.     

二氮嗪预处理联合低温对大鼠海马神经元缺氧复氧时Bcl-2和Bax表达的影响

Objective To investigate the effects of diazoxide preconditioning combined with hypothermia on the expression of Bcl-2 and Bax during anoxia-reoxygenation in rat hippocampal neurons. Methods The hippocampal neurons isolated from newborn SD rats ( < 24 h, weighing 5-6 g) were inoculated in the culture dish or 96 well plates. The hippocampal neurons were randomly assigned into 8 groups and each group contained 36 wells or 12 dishes of neurons: normal temperature .group (group NT), diazoxide preconditioning (DP) + NT group (group DP+ NT), mild hypothermia group (group MiH) , DP + MiH group (group DP + MiH) , moderate hypothermia group (group MoH), DP + MoH group (group DP + MoH), deep hypothermia group (group DeH) and DP+DeH group (group DP+ DeH). In group DP + NT, DP + MiH, DP+ MoH and DP + DeH, diazoxide was added to the culture media, the final concentration was 100 μmol/L,and the neurons were incubated for 1 h once a day for 2 d, and then subjected to 4 h of hypoxia at 37, 34, 30 and 22℃ , respectively, followed by 48 h of reoxygenation at 37℃ . The neuronal viability, apoptotic rates and expression of Bcl-2 and Bax were determined and the ratio of Bcl-2/Bax was calculated. Results The neuronal viability was significantly higher, apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP + NT, MiH, MoH and DeH than in group NT ( P < 0.05). The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP + MiH, DP + MoH and DP+ DeH than in group DP + NT ( P < 0.05), but there was no significant difference in the late apoptotic rate between group DP + MiH, DP + MoH, DP + DeH and DP + NT ( P > 0.05). The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP+ MiH and DeH than in group MiH (P < 0.05), but there was no significant difference in the late apoptotic rate between group DP + MiH, DeH and MiH, and no significant difference in the above indices between group MoH and MiH (P > 0.05) . The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP+ DeH than in group DP+ MiH ( P < 0.05). Conclusion Diazoxide preconditioning combined with hypothermia can attenuate the anoxia-reoxygenation injury in rat hippocampal neurons possibly through correcting the imbalance of Bcl-2 and Bax and inhibiting the early apoptosis of hippocampal neurons.     

含二氮嗪停搏液对豚鼠心肌电生理的影响

目的　观察线粒体内膜ATP敏感钾离子通道开放剂二氮嗪加入St Thomas液内对缺血再灌注后豚鼠心肌的保护作用。方法　 2 4只豚鼠随机分为 3组 ,每组 8只 ,取离体心脏的乳头肌标本 ,充氧台氏液灌注平衡 80min。分别灌注不同的停搏液 ,停搏 30min ,复灌 6 0min。对照组灌注St Thomas液 ;处理组灌注含二氮嗪的St Thomas液 ;阻滞剂组 ,在平衡后 15min加入Glibenclamide ,余同处理组。结果　处理组的复跳时间明显短于其他两组 (P <0 0 5 )。再灌注后处理组的动作电位振幅 (APA)、动作电位超射值 (OS)、动作电位最大除极速度 (Vmax)、5 0 %动作电位时程 (APD50 )及 90 %动作电位时程 (APD90 )的恢复优于对照组 ,差异有显著性 (P <0 0 5 )。处理组复灌早期的APD50 和APD90 明显少于对照组和阻滞剂组。结论　含二氮嗪的St Thomas液能够促进缺血再灌注后心肌功能的恢复。     

二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时PI3K mRNA和Akt mRNA表达的影响

目的 评价二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时磷脂酰肌醇-3激酶(PI3K) mRNA和蛋白质丝氨酸苏氨酸激酶(Akt) mRNA表达的影响.方法 培养SD大鼠心肌微血管内皮细胞,以1×106个/ml密度接种于96孔培养板(100μl/孔)或培养皿(2 ml/皿),采用随机数字表法,将其随机分为4组(n=25).正常对照组(C组)不作任何处理;缺氧复氧组(H/R组)、二氮嗪预先给药组(DZ组)、二氮嗪预先给药+线粒体ATP敏感性钾通道阻断剂5-羟葵酸组(DZ+ 5-HD组)均进行缺氧2h,复氧2h.DZ组和DZ+ 5-HD组在缺氧前2h分别加入100 μmol/L二氮嗪、100μnol/L二氮嗪+ 100 μmol/L 5-羟葵酸.于复氧2h时测定细胞活力、细胞凋亡率、PI3K mRNA和Akt mRNA表达.结果 与C组比较,H/R组细胞活力降低,细胞凋亡率升高,PI3K mRNA和Akt mRNA表达上调(P＜0.05或0.01);与H/R组比较,DZ组细胞活力升高,细胞凋亡率降低,PI3K mRNA和Akt mRNA表达上调(P ＜0.05或0.01);5-羟葵酸可逆转二氮嗪预先给药导致的上述改变(P＜0.05或0.01).结论 二氮嗪预先给药减轻大鼠心肌微血管内皮细胞缺氧复氧损伤的机制与激活线粒体ATP敏感性钾通道,促进PI3K和Akt基因的转录有关.     

二氮嗪预处理联合低温对大鼠海马神经元缺氧复氧时Bcl-2和Bax表达的影响

Objective To investigate the effects of diazoxide preconditioning combined with hypothermia on the expression of Bcl-2 and Bax during anoxia-reoxygenation in rat hippocampal neurons. Methods The hippocampal neurons isolated from newborn SD rats ( < 24 h, weighing 5-6 g) were inoculated in the culture dish or 96 well plates. The hippocampal neurons were randomly assigned into 8 groups and each group contained 36 wells or 12 dishes of neurons: normal temperature .group (group NT), diazoxide preconditioning (DP) + NT group (group DP+ NT), mild hypothermia group (group MiH) , DP + MiH group (group DP + MiH) , moderate hypothermia group (group MoH), DP + MoH group (group DP + MoH), deep hypothermia group (group DeH) and DP+DeH group (group DP+ DeH). In group DP + NT, DP + MiH, DP+ MoH and DP + DeH, diazoxide was added to the culture media, the final concentration was 100 μmol/L,and the neurons were incubated for 1 h once a day for 2 d, and then subjected to 4 h of hypoxia at 37, 34, 30 and 22℃ , respectively, followed by 48 h of reoxygenation at 37℃ . The neuronal viability, apoptotic rates and expression of Bcl-2 and Bax were determined and the ratio of Bcl-2/Bax was calculated. Results The neuronal viability was significantly higher, apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP + NT, MiH, MoH and DeH than in group NT ( P < 0.05). The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP + MiH, DP + MoH and DP+ DeH than in group DP + NT ( P < 0.05), but there was no significant difference in the late apoptotic rate between group DP + MiH, DP + MoH, DP + DeH and DP + NT ( P > 0.05). The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP+ MiH and DeH than in group MiH (P < 0.05), but there was no significant difference in the late apoptotic rate between group DP + MiH, DeH and MiH, and no significant difference in the above indices between group MoH and MiH (P > 0.05) . The neuronal viability was significantly higher, early apoptotic rate lower, Bcl-2 expression higher, Bax expression lower and Bcl-2/Bax ratio higher in group DP+ DeH than in group DP+ MiH ( P < 0.05). Conclusion Diazoxide preconditioning combined with hypothermia can attenuate the anoxia-reoxygenation injury in rat hippocampal neurons possibly through correcting the imbalance of Bcl-2 and Bax and inhibiting the early apoptosis of hippocampal neurons.     

二氮嗪治疗先天性高胰岛素血症7例

目的 分析二氮嗪试验性治疗先天性高胰岛素血症(CHI)的疗效.方法 回顾性分析2006-2008年本院收治并经实验室检查诊断为CHI,并试用二氮嗪治疗的7例患儿(男5例;女2例;起病年龄2 d～4个月;出生体质量2.65～4.60 kg)的临床资料,并对其方法及疗效进行分析.结果 7例患儿确诊CHI后,均应用二氮嗪进行7～10 d的试验性治疗.其中5例均为新生儿期以后起病,经治疗后,血糖逐渐恢复至正常水平,对二氮嗪治疗有效.2例分别于出生2 d和出生1个月起病,治疗10 d后,仍呈现严重而持续的低血糖症,对二氮嗪治疗无效.结论 因部分CHI患儿的细胞具有功能正常的ATP敏感性钾通道,应用二氮嗪治疗有效,因此对确诊CHI的患儿应首先试用二氮嗪治疗.     

二氮嗪对氯化锂-匹鲁卡品致痫大鼠海马神经元超微结构及自由基的影响

目的 观察线粒体ATP敏感性钾通道(mitoKATP)开放剂二氮嗪(DZ)对氯化锂-匹鲁卡品致痫大鼠海马神经元超微结构及自由基的影响,探讨mitoK开放剂对癫痫发作后神经元的保护机制.方法 随机将成年雄性Wistar大鼠80只,分为对照组、癫痫组(PILO组)、DZ组(DZ组)、DZ+5-羟基癸酸(5-HD)...     

奥曲肽治疗先天性高胰岛素血症1例报道

<正>患儿,男,54 d,因发现血糖低51 d入院。患儿51 d前(生后3 d)无明显诱因于睡眠中出现惊厥,于当地医院就诊,发现患儿存在严重低血糖(最低0.9 mmol/L),血清胰岛素水平明显升高(50～281μIU/mL),诊断为"顽固性低血糖,高胰岛素血症",因诊治效果欠佳,后转来我院,以"低血糖原因待查"收入院治疗。     

二氮嗪预处理对自发性高血压大鼠心肌缺血再灌注损伤的影响及机制

目的　探讨二氮嗪在离体自发性高血压大鼠心脏上模拟缺血预处理效应的可能性及线粒体KATP在其中的作用。方法　雄性自发性高血压大鼠取心 ,行Langendorff灌流。实验分为 5组 (n =6 ) :对照组 (Con)在平衡后继续灌流 4 0min ,全心缺血 2 5min ,复灌 30min ;其余各组除全心缺血前处理不同外 ,余均同对照组。缺血预处理组 (IP组 ) 2次给予 5min缺血 +10min复灌 ;二氮嗪预处理组 (DP组 )给予 2次含 5 0 μmol·L- 1 二氮嗪的K -H液10min后给不含二氮嗪的K -H液 5min ;5 -HD、5 -HD +DP组则在平衡后给予 10min 15 0 μmol·L- 1 线粒体KATP(mitoKATP)阻断剂 5 -HD ,余同Con组及DP组。结果　IP组及DP组复灌末左室发展压、左心室最大上升速率 (+dP dtmax)和左心室最大下降速率 (-dP dtmax)均高于Con组 (P <0 .0 1) ,但 2组左室舒张末期压低于Con组 (P <0 .0 1) ;5 -HD能拮抗二氮嗪引起的心功能指标的改善。IP组和DP组冠脉流量高于Con组 ,而MDA含量低于Con组 (P <0 .0 1) ,5 -HD组和Con组无差异。结论　二氮嗪预处理对离体自发性高血压大鼠心肌缺血再灌注损伤有保护作用 ,mitoKATP拮抗剂 5 -HD能拮抗此作用