血红素加氧酶-1对TNF-α引起内皮细胞炎症损伤的保护作用

目的探讨血红素加氧酶-1(HO-1)在肿瘤坏死因子-α(TNF-α)导致肺微血管内皮细胞损伤中的保护作用。方法采用TNF-α刺激人肺微血管内皮细胞(HPMECs)模拟重症急性胰腺炎肺损伤的体外模型,锌原卟啉-IX(ZNPP-IX)作为HO-1抑制剂预处理细胞。试验分为对照组、TNF-α组、ZNPP-IX组。CCK8比色法检测细胞活性,采用Western blotting法及RT-PCR法检测HO-1、细胞间黏附分子-1(ICAM-1)的表达,黏附试验检测HPMECs对多形核细胞(PMN)的黏附力。结果 1与对照组相比,TNF-α组(78.69%±5.54%)、ZNPP-IX组(62.00%±4.27%)细胞活性明显降低(P0.01);2与对照组相比,TNF-α组(1.59±0.19)HO-1表达增高(P0.05),ZNPP-IX组(0.01±0.01)比TNF-α组显著降低(P0.01);3与对照组相比,TNF-α组(32.72±0.95)、ZNPP-IX组(85.33±2.37)ICAM-1表达明显升高(P0.01),且ZNPP-IX组比TNF-α组更显著(P0.01);4TNF-α引起HPMECs对PMN黏附力增高,抑制HO-1表达后,黏附作用增强。结论 HO-1可能通过下调ICAM-1的表达降低炎症时HPMECs对PMN的黏附,从而改善重症急性胰腺炎引起的肺损伤。     

香菇多糖对谷氨酸损伤原代培养大鼠神经细胞保护作用的研究

目的观察香菇多糖（Lentinan，LNT）对谷氨酸（glutamate，Glu）损伤体外原代培养大鼠皮层神经细胞的保护作用并探讨可能的机制。方法体外培养胚胎大鼠大脑皮层神经细胞，建立Glu损伤模型，观察LNT的保护作用。结果LNT能显著提高Glu损伤神经细胞的生存率，降低乳酸脱氢酶（LDH）的漏出量，一氧化氮（No）含量及丙二醛（MDA）的生成，提高超氧化物歧化酶（SOD）的活性。结论LNT对Glu损伤神经细胞有显著的保护作用，其机制可能与拮抗Glu诱导的氧化应激有关。     

新型组蛋白去乙酰化酶抑制剂对缺氧损伤心肌细胞的保护作用研究

目的：利用化学发光方法和细胞筛选模型,检测新型组蛋白去乙酰化酶抑制剂（ histone deacetylase inhibitor, HDACi）JZ005的抑制组蛋白去乙酰化酶（histone deacetylases,HDACs）的活性;建立氯化钴损伤的心肌细胞缺氧模型,初步探讨JZ005对缺氧损伤细胞的保护作用。方法采用脂质体转染法将含有p21启动子元件的荧光素酶报告基因真核表达载体pCI-p21-Luc转入到人胚肾细胞293中,用G418筛选获得稳定转染荧光素酶报告基因的单克隆细胞系;采用已报道的HDACi曲古抑菌素A（ trichostatina A,TSA）为阳性对照,检测细胞筛选模型的稳定性;用HDACi化学发光检测试剂盒及上述细胞筛选模型测定JZ005抑制HDACs的活性;用不同浓度的JZ005处理氯化钴缺氧损伤的大鼠胚胎心肌细胞（H9c2）,MTT法检测JZ005对缺氧损伤细胞的保护作用。免疫印迹法检测JZ005处理后正常及缺氧损伤心肌细胞组蛋白H3的乙酰化水平变化。流式细胞术检测JZ005对H9c2细胞缺氧损伤后凋亡的影响。结果建立含p21启动子元件荧光素酶报告基因的HDACi细胞筛选模型;JZ005能够显著抑制HDACs的活性,浓度50～400μmol／L,抑制率＞50％。对缺氧损伤的心肌细胞具有明显保护作用,与对照组相比,细胞存活率提高38．33％、56．00％和35．20％,同时能够上调缺氧损伤心肌细胞组蛋白H3的乙酰化水平,拮抗缺氧损伤心肌细胞的凋亡,细胞凋亡数目从对照组的12．89％分别下降到给药组（25,50和100μmol／L）的6．63％、10．56％和8．89％。结论成功建立了HDACi的细胞筛选模型;JZ005作为一种新型的HDACi ,具有明显的保护心肌细胞拮抗缺氧损伤的作用,提示JZ005有可能开发成一种治疗缺氧损伤的药物。     

星状神经节阻滞对家兔急性肺损伤外周血中炎性细胞因子的影响

目的 探讨星状神经节阻滞(SGB)对急性肺损伤(ALI)家兔炎性反应的影响,从免疫调节角度阐明SGB的肺保护作用.方法 30只健康成年家兔随机分成三组(n=10):盐水对照组(A组)、盐酸模型组(B组)和SGB干预组(C组).A组气管内给1.5 ml/kg0.9％氯化钠注射液；B组气管内给予盐酸1.5 ml/kg建立吸入性肺损伤模型；C组模型建立后给予0.25％布比卡因行右侧星状神经节阻滞.测定建模后6h外周血中IL-2、IL-4、IL-6及IL-10含量；取肺组织测湿干重比(G/T).结果 与A组比较,B组血清中IL-2和IL-6含量显著升高(P =0.037,P=0.043)；与B组比较,C组血清中IL-2和IL-6含量降低(P =0.035,P=0.043)；与A组比较,B组血清中IL-4和IL-10含量显著降低(P =0.032,P=0.045)；与B组比较,C组血清中IL-4和IL-10含量增加(P =0.033,P=0.046)；与A组比较,B组G/T值显著升高(P=0.045)；与B组比较,C组G/T值显著降低(P =0.048).结论 星状神经节阻滞可调节盐酸致家兔急性肺损伤外周血中细胞因子蛋白表达,减轻炎性反应,发挥肺保护作用.     

全身麻醉药的临床新用途及其作用机制研究进展

全身麻醉药主要通过正向调控中枢γ-氨基丁酸A型受体使意识可逆性消失,是外科手术中达到理想全身麻醉状态的必须药物。随着全身麻醉药临床应用经验的积累以及新研究技术、方法的发展,其具有的一些潜在临床应用方向和线索被发现,如器官保护作用、抗肿瘤作用、抗精神病作用和抗癫痫作用。不同的全身麻醉药在药效作用和临床应用方向上存在差异,对其作用机制的深入研究和比较,不仅可为临床上合理用药提供科学性依据,也可进一步针对关键作用靶点进行药物优化和创新。     

非酒精性脂肪性肝炎大鼠血红素加氧酶-1的表达与氧应激的关系

非酒精性脂肪性肝病是一种常见肝脏疾患,有逐年增加的趋势。近年的研究认为氧应激和脂质过氧化反应是其发病机制的轴心[1]。血红素加氧酶(HO)是血红素降解的起始酶和限速酶,HO-1为其诱导型。众多证据表明,各种体内及体外氧化性损伤模型中HO-1提供了细胞保护作用,因而认为HO-1的诱导是机体对抗氧应激的一种代偿机制[2]。本文在建立大鼠高脂饮食脂肪性肝炎模型的基础上,探讨非酒精性脂肪性肝炎中HO-1的表达情况及其与氧应激的关系,为进一步研究HO-1在非酒精性脂肪性肝炎中的作用提供依据。材料与方法一、动物模型的建立和分组雄性Wistar…     

黄芪甲苷对过氧化氢致损心肌细胞的保护作用及机制研究

目的 探讨黄芪甲苷对过氧化氢致损心肌细胞的保护作用及机制.方法 选取新生Wistar 大鼠3只,体外分离大鼠原代心肌细胞,按0.6×106/ml密度接种至75 cm2培养瓶培养,待80%融合后予0.5 mmol/L过氧化氢培养基培养4 h,分别加入100、10、1、0.1 μmol/L黄芪甲苷培养基6 ml培养24 h;设立对照组,仅加入等量普通培养基.观察心肌细胞存活率,乳酸脱氢醇(LDH)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)酶活性情况,bax和bcl-2基因的表达,bax和bcl-2蛋白的表达情况.结果 与不同浓度黄芪甲苷共培养24 h的过氧化氢致损心肌细胞的成活率(0.1、1、10、100 μmol/L黄芪甲苷组分别为48%、55%、57%、52%)均明显高于对照组(30%),(均P＜0.01);黄芪甲苷可降低H2O2致损心肌细胞内的bax mRNA和蛋白的表达,提高bcl-2 mRNA和蛋白的表达.结论 黄芪甲苷对过氧化氢致损心肌细胞有保护作用,其机制可能是通过调节bax/cbl-2基因和蛋白的表达实现的.

Abstract：

Objective To study the effects of Astragaloside Ⅳ on injured rat cardiomyocytes caused by hydrogen peroxide (H2O2). Methods Three clean Wistar rats were supplied by Experimental Animal Center,Xingjiang medical university. Primary rat cardiomyocytes were isolated in vitro and cultured in 75 cm2 flasks. When cells were at 80% confluence, they were cultured with 0.5 mmol/L H2O2 medium for 4 hours and then treated with 6 ml 100, 10, 1,0. 1 μmol/L Astragaloside Ⅳ medium for24 hours. Cells in the control group were treated with an equal volume of normal medium. Cell survival rate were examined by trypan blue exclusion assay. Enzyme activities of lactate dehydrogenase(LDH), superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) were assayed by Elisa. The mRNA and protein expression were detected with realtime polymerase clain reaction(PCR) and Westernhlot method separately. Results Astragaloside Ⅳ promoted cardiomyocytes survival rate in a dose-dependent manner, while high concentration showed an inhibition tendency. Astragaloside Ⅳ decreased LDH activity (P ＜0.01 ) but increased SOD and GSH-Px activities (P ＜0.05 ). The results of Realtime PCR and Westernblot showed that Astragaloside Ⅳ could suppress expression of bax mRNA and protein, promote expression of cbl-2 mRNA and protein. Conclusions Astragaloside Ⅳ has protective effects on injured rat cardiomyocytes caused by H2O2, which is probably associated with regulating expression of bax/bcl-2 gene and protein.     

阿魏酸钠对婴幼儿体外循环血管内皮功能的保护作用

Objective Cardiopulmonary bypass (CPB) and its related ischemia reperfusion injury may cause endothelial cell injury.To study the protective effects of sodium ferulate in vascular endothelial function during CPB by testing the changes of vascular endothelial cell( CEC),nitric oxide( NO) and endothelin-1 ( ET-1 ) in children with congenital heart disease.Methods Sixty patients with congenital heart disease,including 28 males and 32 females were studied.The mean age was (19.7 ±10.4) months and body weight (10.5 ±6.1) kg.There were 37 VSD,8 ASD,7 TOF,5 TAPVC and 3 CAVC,among them 26 patients had pulmonary hypertension.They were randomly divided in to two groups:sodium ferulate group ( group S,n = 30),and control group ( group C,n =30) .Sodium ferulate (8 mg/kg) was given intravenously before CPB.Blood samples were taken from the arterial line at following time points:before CPB (TO),bypass 30 min(Tl ),the termination of CPB (T2 ),2h after operation ( T3 ) and 6h after operation ( T4 ),respectively for determination the concentration of vascular endothelial cell (CEC) in the blood,the concentration of nitric oxide (NO) and endothelin-1 ( ET-1) in the plasma.Results There were no significant difference for the two groups regarding above parameters at TO ( P > 0.05).The level of CEC was significantly elevated after CPB in both groups ( P < 0.05 ) .CEC were lower at T2 in group S than in group C ( P < 0.05 ) .NO was decreased in both groups,but was higher in group S at T2,T3 and T4 ( P < 0.05 ) .The concentration of plasma ET-1 was not significantly different before CPB,but there was a slight decrease at T1,and then it was significantly increased in both groups (P<0.05).But it was lower in group S than in group C at T1,T2,T3 and T4(P<0.05 orP<0.01).Conclusion There was severe endothelial cell damage during CPB.Sodium Ferulate can effectively antagonize the secretion of ET-1 to promote the formation of NO.Therefore,it reduces CPB-induced endothelial cell damage and protects vascular endothelial function during CPB.     

硫化氢对PC12细胞存活素表达的影响及其神经保护作用

Objective To explore the effect of hydrogen sulfide (H2S) on the expression of survivin in PC12 cells and the neuroprotective function of H2S on PC12 cells.Methods Different concentrations of sodium hydrosulfide (NaHS) were used to treat the PC12 cells at different times.Dose-effect (50-800 μmol/L) and time-effect (0-180 min) on the expression of survivin were evaluated by Western blotting.Cell viability was tested by using cell counter kit-8.Results NariS treatment at the concentrations from 50 to 200 μmol/L for 30 min could up-regulate the expression of survivin in a dose dependent manner,however,when the concentration of NariS was above that,the expression of survivin decreased gradually;when the concentration of NariS reached 800 μmoi/L,the expression level of survivin was lower than the normal level.Treatment with 400 μmol/L NariS within the range of 0-60 min could promote the expression of survivin in a time dependent manner,but with the extension of time,the expression of survivin was declined.On the other hand,400 μmol/L NaHS preconditioning could enhance the expression of survivin promoted by CoCl2 and reduce the injuries of PC12 cells induced by CoCl2 to increase the cell viability.Conclusion H2S increases the expression ofsurvivin in a dose and time dependent manners at certain degree,which may be related to the protection of PC12 cells against chemical hypoxic damage.