牛磺酸对外源性羟自由基诱导心肌细胞凋亡的影响

目的：观察牛磺酸对外源性羟自由基诱导心肌细胞凋亡的影响。方法：采用培养的第2代心肌细胞，随机分面4组：（1）正常对照组：仅用DMEM培养；（2）羟自由基组：OH－终浓度为0．1mmol.L^-1；（3）牛左酸组：牛磺酸终浓度为30mmol.L^-1；（4）牛磺酸＋羟自由基组：牛磺酸30mmol.L^-1 OH-0.1mmol.L。观察心肌细胞存活率和形态学，流式细胞仪双标法检测细胞凋亡率。结果：（1）羟自由基组心肌细胞存活率降低，和对照组比较有显性差异（P＜0．05），牛磺酸＋羟自由基组细胞存活率较羟自由基组高（P＜0．05），（2）Annexin V /PI-细胞即凋亡细胞在羟自由基组有较高的发生率，明显高于其他各组（P＜0．05）；牛磺酸＋羟自由基组的细胞凋亡率显低于羟自由基组（P＜0．05），（3）羟自由基组凋亡心肌细胞呈特征性超微结构。结论：牛磺酸能减轻外源性羟自由基诱导的心肌细胞凋亡。     

卡托普利对外源性羟自由基诱导心肌细胞凋亡的影响

目的观察卡托普利对由外源性羟自由基诱导的心肌细胞凋亡有无影响。方法①利用第４代心肌细胞,随机分１５组,在终浓度为１０＾－５ｍｏｌ／Ｌ、１０＾－４ｍｏｌ／Ｌ、１０＾－３ｍｏｌ／Ｌ、１０＾－２ｍｏｌ／Ｌ、１０＾－１ｍｏｌ／Ｌ的羟自由基无血清条件培养下分别共孵育８ｈ、１６ｈ、２４ｈ,确定１０＾－３ｍｏｌ／Ｌ及２４ｈ为诱导心肌细胞凋亡的最佳浓度及时间。②在上述条件下分别观察３种不同浓度卡托普利（１０＾－７     

卡维地洛对外源性羟自由基诱导心肌细胞凋亡的影响

目的 :观察卡维地洛 (Carvedilol)对外源性羟自由基诱导心肌细胞凋亡的影响。方法 :采用培养的第 2代心肌细胞 ,随机分成 4组 :1.正常对照组 :仅用DMEM培养 ;2 .羟自由基组 :FeSO410 0 μmol·L- 1 +H2 O2 10 0 μmol·L- 1 ;3.卡维地洛组 :卡维地洛终浓度为 1μmol·L- 1 ;4 .卡维地洛 +羟自由基组 :卡维地洛 1μmol·L- 1 +FeSO4 10 0 μmol·L- 1 +H2 O2 10 0 μmol·L- 1 。观察心肌细胞存活率和形态学 ,流式细胞仪检测细胞凋亡率。结果 :1.羟自由基组心肌细胞存活率降低 ,和对照组比较有显著性差异(P <0 0 5 ) ;卡维地洛 +羟自由基组细胞存活率较羟自由基组高 (P <0 0 5 )。 2 .羟自由基组凋亡率较高 ,明显高于对照组 (P <0 0 5 ) ;卡维地洛 +羟自由基组的细胞凋亡率显著低于羟自由基组 (P <0 0 5 )。3.羟自由基组凋亡心肌细胞呈特征性超微结构及AnnexinV+ PI- 荧光染色。结论 :卡维地洛能减轻外源性羟自由基诱导的心肌细胞凋亡。     

胺碘酮对外源性羟自由基诱导心肌细胞凋亡的影响

目的　观察胺碘酮对外源性羟自由基诱导心肌细胞凋亡的影响。方法　采用培养的第 2代心肌细胞 ,随机分成 4组 :1正常对照组 :仅用 DMEM培养 ;2羟自由基组 :OH-终浓度为 0 .1 mmol/ L;3胺碘酮组 :胺碘酮终浓度为 1 0μmol/ L;4胺碘酮 +羟自由基组 :胺碘酮 1 0μmol/ L+OH- 0 .1 mmol/ L。观察心肌细胞存活率和形态学 ,流式细胞仪双标法检测细胞凋亡率。结果　 1羟自由基组心肌细胞存活率降低 ,与对照组比较有显著性差异 (P<0 .0 5) ;胺碘酮 +羟自由基组细胞存活率较羟自由基组高 (P<0 .0 5)。 2 Annexin V+ / PI-细胞即凋亡细胞在羟自由基组有较高的发生率 ,明显高于其他各组 (P<0 .0 5) ;胺碘酮 +羟自由基组的细胞凋亡率显著低于羟自由基组 (P<0 .0 5)。 3羟自由基组凋亡心肌细胞呈特征性超微结构及 Annexin V+ / PI-荧光染色。结论　胺碘酮能减轻外源性羟自由基诱导的心肌细胞凋亡。     

脑缺血再灌注肾脏组织损伤老年大鼠ATP酶和自由基代谢的变化及其意义

目的 从ATP酶活性变化和自由基损伤方面研究老年大鼠脑缺血再灌注肾脏损伤机制。方法 青年（5-6月龄）和老年（20-22月龄）大鼠均分为模型组和正常对照组。观察大鼠全脑缺血30min再灌注60min后肾脏组织形态和肌酐（Cr)。尿素氮（BUN），丙二醛（MDA）含量及超氧化物岐化酶（SOD），ATP酶的活性，结果 青年和老年模型组大鼠肾脏组织形态和功能均出现明显的病理改变，老年模型组较青年模型组严重，青年模型组和老年模型组肾脏组织MDA含量和MDA/SOD比值分别高于青年对照组和老年对照组，老年模型组肾脏组织Na^ -K^ -ATP酶和Ca^2 -ATP酶活性低于青年模型组；老年对照组肾脏Ca^2 -ATP酶活性低于青年对照组，老年模型组肾脏Ca^2 -ATP酶活性低于老年对照组。结论 脑缺血再灌注肾脏损伤老年大鼠较青年大鼠严重。ATP酶活性降低和自由基损伤可能是其主要机制之一，由于老年大鼠ATP酶活性和自由基代谢的增龄变化使这些病理改变较青年明显并具有一定特点。     

老年大鼠脑缺血再灌注心肌组织ATP酶和自由基代谢的变化及其意义

目的：从ATP酶活性变化和自由基损伤方面研究老龄大鼠脑缺血再灌注心肌损伤机制。方法：青年（5月龄）和老年（20月龄以上）大鼠均分为模型组和正常对照组,观察大鼠全脑缺血30分钟再灌注60分钟后ATP酶、乳酸脱氢酶（LDH）、肌酸磷酸激酶（CPK）、超氧化物歧化酶（SOD）的活性和丙二醛（MDA）含量。结果：血清中CPK和LDH水平青年模组明显高于青年对照组和老龄模型组,老龄模型组高于老龄对照组;老龄对照组血清中CPK显高于青年对照组。老龄模型组心肌Na^ -K^ -ATP酶活性和青年对照组Ca^2 -ATP酶活性较老龄对照组和青年模型组均降低;老龄模型组心肌组织MDA／SOD比值高青年模型组和老龄对照组。结论脑缺血再灌注心肌损伤可能与ATP酶活性的变化和自由基损伤有关。但由于老龄大鼠ATP酶活性和自由基代谢的增龄变化使脑缺血再灌注后这些病理性变青年大鼠明显并具有一定特点。     

卡托普利对再灌注损伤心肌细胞LDH和MDA的影响

目的 :观察卡托普利对大鼠心室肌细胞缺氧 -复氧损伤的影响。方法 :酶解法分离心肌细胞 ,将实验分为 9组 :不缺氧组、单纯缺氧组、缺氧 -复氧组、卡托普利 0 .0 5、0 .1、0 .2、0 .4、0 .8和 1.6μmol/L灌流组。观察杆形心肌细胞百分比、测定乳酸脱氢酶 （L DH）活性、L DH活性百分比和丙二醛 （MDA）含量。结果 :不缺氧组心肌细胞的生存率、释放 L DH活性百分比、MDA含量无明显变化。单纯缺氧组心肌细胞的生存率下降 ,释放 L DH活性百分比明显增加 ,MDA的含量亦增加 ;复氧组较缺氧组心肌细胞造成更严重的损伤 （P<0 .0 1） ;卡托普利 0 .0 5,0 .1μm ol/L 对缺氧 -复氧造成的损伤无明显作用 （P>0 .0 5） ,但 0 .2 ,0 .4,0 .8和 1.6μm ol/L能明显提高心肌细胞缺氧 -复氧过程中的生存率、减少脂质过氧化产物的生成 ,且呈剂量依赖性。结论 :卡托普利对缺氧 -复氧损伤的心肌细胞具有剂量依赖性保护作用     

Staurosporine诱导小鼠心肌细胞凋亡模型的构建

目的 :Staurosporine可诱导不同细胞类型的细胞凋亡 ,但尚缺乏其对小鼠心肌细胞作用影响的报道。本研究旨在观察 Staurosporine是否也可引起小鼠心肌细胞的凋亡 ,并对其模型进行分子水平的鉴定。方法 :用 Stau-rosporine处理原代培养的小鼠心肌细胞的直接刺激后 ,观察心肌细胞的形态学变化、DNA片段、半胱天冬蛋白酶(caspase) -3活性、细胞存活率以及细胞膜完整性。结果 :1Staurosporine处理过的心肌细胞具有明显的凋亡形态学特征 :细胞的皱缩、核的浓缩及 DNA梯改变。2 Staurosporine4μmol/ L 对心肌细胞作用 8h后 ,与正常对照组比较细胞存活率降至 3 1.2 % (与对照组比 ,P<0 .0 1) ,且细胞存活率与 Staurosporine的作用浓度和作用时间呈依赖关系 ;同时测定细胞培养液中能反映细胞膜完整性的胞浆内容物乳酸脱氢酶 (L DH)水平 ,发现在 Staurosporine作用 1～ 8h的各个时间点 ,L DH的漏出水平最高未超过 10 % (与对照组相比 ,P>0 .0 5) ;这种高的细胞死亡率与低的细胞膜的损伤率 ,恰恰反映了细胞死亡的性质是凋亡性死亡。3 Staurosporine能激活心肌细胞内的 Caspese-3的活性 ,当使用广谱的 Caspese抑制剂 ZVAD-fmk预处理培养的小鼠心肌细胞后 ,能够有效的阻止上述细胞凋亡的发生 ,从而避免了 Staurospor     

心力衰竭时红细胞内外离子及ATP酶变化及临床意义

近年来从细胞分子水平探讨心肌细胞钠、钾、钙、镁离子及ＡＴＰ酶的变化规律及在某些疾病的变异与临床意义，已引起人们的日益关注。研究资料证实心肌细胞与红细胞离子的含量、ＡＴＰ酶的变化基本上是一致的，因此常以研究红细胞离子的代谢变化以反映（或替代）心肌细胞离子的代谢变化。这里简要讨论正常人体及老年心力衰竭时红细胞钠、钾、钙、镁离子及ＡＴＰ酶的变化与临床意义。１　生理状况下钠、钾、钙、镁离子的分布　　生理状态下细胞内钾占９８％～９８－５％，细胞外液钾仅１－５％～２％；钠在细胞内占１０％，细胞外液占５０％，…     

肉桂水提液对全脑缺血再灌注损伤大鼠羟自由基和谷胱甘肽过氧化物酶的影响

目的 探讨肉桂水提液对全腩缺血再灌注损伤大鼠羟自由基(·OH)和谷胱甘肽过氧化物酶(GSH-Px)活性的影响.方法 采用4-血管阻断(4-VO)方法建立全脑缺血再灌注动物模型,利用分光光度法测定大鼠脑、肝、肾、心等组织中·OH抑制能力和GSH-PX活性的变化.结果 与模型组相比,各剂量给药组大鼠各组织中·OH的抑制能力和谷胱甘肽过氧化物酶(GSH-PX)的活性明显提高(P<0.01或P<0.05).结论 肉桂水提液可预防性降低自由基代谢造成的机体损伤,提高GSH-Px的活性,对脑缺血再灌注损伤具有保护作用.     

Alterations in heart sarcolemmal Ca2+-ATPase and Ca2+-binding activities due to oxygen free radicals

Summary Effects of oxygen free radicals on Ca²⁺/Mg²⁺ ATPase and ATP-independent Ca²⁺-binding activities were examined in rat heart sarcolemma. Membranes were incubated with different oxygen radical generating media such as xanthine + xanthine oxidase, hydrogen peroxide, and hydrogen peroxide + Fe²⁺. In the presence of xanthine + xanthine oxidase, Ca²⁺ ATPase activity was stimulated and this effect was prevented by the addition of superoxide dismutase. Hydrogen peroxide also showed a significant increase in Ca²⁺-ATPase activity in a dose-dependent manner and this effect was blocked by catalase. On the other hand, a combination of hydrogen peroxide + Fe²⁺ decreased Ca²⁺-ATPase activity; this depression was prevented by the addition of D-mannitol. The observed change in Ca²⁺-ATPase activity due to oxygen free radicals was associated with changes in V_max, whereas Ka remained unaffected. Both xanthine + xanthine oxidase and hydrogen peroxide increased whereas, hydrogen peroxide + Fe²⁺ inhibited the ATP-independent Ca²⁺-binding activities. It is suggested that oxygen free radicals may influence Ca²⁺ movements in the cell by altering the Ca²⁺/Mg²⁺ ATPase and Ca²⁺-binding activities of the membrane and these effects may be oxygen-radical species specific.     

心肌肌浆网钙转运蛋白结构、功能调节及基因表达

本文简要综述了心肌肌浆网的结构和功能 ,重点综述钙转运蛋白的结构、功能及其调节和基因表达     

Alterations of Na+-K+-ATPase, Ca2+-ATPase, and Mg2+-ATPase activities in erythrocyte, muscle, and liver of traumatic and septic patients

Na+-K+-ATPase, Ca2+-ATPase and Mg2+-ATPase activities of erythrocyte membrane, microsomal fractions of rectus muscle, and liver were measured colorimetrically in the biopsy specimens of 14 control, 7 uncomplicated trauma (group 2), and 14 severe trauma or septic patients (groups 3-A and 3-B). In erythrocytes, these three ATPase activities in group 2 were not significantly changed but sepsis of both the acute (group 3-A) and ongoing type (group 3-B) decreased all of the ATPase activities. In muscle, there was a significant loss of three ATPase activities in the acute insult of severe trauma or sepsis (group 3-A), while Na+-K+-ATPase and Mg2+-ATPase activities were not significantly changed in ongoing, severe trauma (group 3-B). In the liver, a tendency for all three ATPase activities to decrease is noted in the severe traumatic group. However, a statistical difference between the control and severe traumatic group showed only for Na+-K+ ATPase and Mg2+-ATPase in group 3-A and Ca2+-ATPase in group 3-B. Correlation coefficients between erythrocyte, muscle, and liver for three ATPase activities are between 0.4 and 0.5. The mechanism which alters ATPase activity remains unknown in this study, but it may account for the variation in traumatic insult, in hemodynamic and hormone changes, and in tissue energy stores.     

肝硬化红细胞钠钾ATP酶、钙镁ATP酶及钠钾钙镁改变

目的:探讨肝硬化时细胞内钠钾钙镁的改变及细胞膜钠钾ATP酶(NKA)、钙镁ATP酶 (CMA)活性改变在细胞内钠钾钙镁改变中的作用．方法:测定了52例肝硬化失代偿期(实验组 A)、36例代偿期(实验组B)患者红细胞及血清钠钾钙镁(RNa、RK、RCa、RMg;SNa、 SK、SCa、SMg)含量和NKA和CMA活性．以 36名健康人为对照组．结果:与对照组比较,实验组A的NKA、 CMA、RK、RMg(t=5．92,P<0．001;t=7．21, P<0．001;t=2．32,P<0．02;t=4．79,P<0．001)和买验组B的NKA、CMA、RK、RMg(t=3．83, P<0．001;t=2．53,P<0．02;t=2．03,P<0．05;t= 3．33,P<0．002)均显著降低;与实验组B比较, 实验组A的NKA、CMA活性(t=2．29,P<0．05; t=4．14,P<0．005)显著降低．与对照组比较, 买验组A的SNa、SK、SCa、SMg(t=8．25, P<0．001;t=5．73,P<0．001;t=9．82,P<0．001; t=6．15,P<0．001)显著降低;与实验组B比较, 买验组A的SNa、SK、SCa、SMg(t=6．94, P<0．001;t=5．00,P<0．001;t=5．57,P<0．001; t=5．73,P<0．001)显著降低．与Child B级组比较,Child C级组的NKA、CMA、RK、RMg、 SNa、SK、SCa、SMg(P<0．05或P<0．01) 显著降低．与非肝性脑病组比较,肝性脑病组NKA、CMA、RK、RMg、SNa、SK、 SMg(P<0．05或P<0．01)显著降低．实验组A中, 低SMg者的NKA和CMA显著低于高SMg者 (16．87±3．19 vs 19．04±3．25;109．83±13．51 vs 120．13±13．27;P均<0．05)．结论:肝硬化患者存在缺钾缺镁,且随病情加重而加重,缺钾缺镁可能为病情加重的原因之一．NKA和CMA活性降低可导致细胞内低钾低镁和钠钙蓄积．缺镁为ATP酶活性在失代偿期进一步降低的原因之一．     

Role of Superoxide and hydroxyl radicals in rat gastric mucosal injury induced by ethanol

It has been reported that oxygen-derived free radicals play an important role in the pathogenesis of mucosal injury in the small intestine as well as in the stomach. The aims of this study were to test whether ethanol-induced damage in the rat stomach was prevented by the administration of (1) Superoxide dismutase (SOD; a scavenger of Superoxide radicals), (2) allopurionol (ALP; an inhibitor of xanthine oxidase), (3) dimethyl sulfoxide (DMSO; a scavenger of hydroxyl radicals). SOD significantly decreased the ulcer index from 100±8.5% (control) to 39.6±8.2% (P<0.001). Ethanol-induced damage was reduced by the administration of ALP by 37.4% (P<0.01). DMSO also diminished the ulcer index from 100±8.5% (control) to 31.6±5.8% (P<0.01). Histochemical studies supported these results. A scanning EM study, however, revealed that surface epithelial cells were not protected by SOD against ethanol-induced damage. These results demonstrated that SOD, ALP and DMSO had the ability to protect gastric mucosa against ethanol-induced injury. Accordingly, oxygen-derived free radicals may be involved in the pathogenesis of ethanol-induced gastric mucosal damage. Surface epithelial cells, however, were not protected even by SOD against ethanol-induced injury. This paper was presented in 87th Annual Meeting of American Gastroenterological Association (1986).     

芩连合剂对实验性胃溃疡大鼠胃黏膜细胞内钙及氧自由基的影响

[目的]探讨芩连合剂对胃溃疡(GU)的保护作用及其作用机制。[方法]采用无水乙醇灌胃法复制大鼠急性GU模型,测定胃黏膜细胞内丙二醛(MDA)、超氧化物歧化酶(SOD)水平、谷胱甘肽过氧化物酶(GSH-Px)活力及钙离子(Ca2 )浓度。[结果]模型组大鼠胃黏膜细胞内Ca2 浓度及MDA表达较其他各组明显升高,SOD、GSH-Px的表达明显降低,差异均有统计学意义(均P<0.01)。[结论]芩连合剂通过降低胃黏膜细胞内Ca2 浓度及MDA的表达,提高SOD和GSH-Px的表达,起到保护胃黏膜的作用,从而发挥其抗GU的作用。     

Mechanisms of Ca2+ overload induced by extracellular H2O2 in quiescent isolated rat cardiomyocytes

Rat cardiomyocytes were exposed to H₂O₂ (1–100 μmol/L) for 10 min with washout for 10 min. Intracellular Ca²⁺ concentration ([Ca²⁺]_i) was measured using fluo-3. [Ca²⁺]_i increased with 100 μmol/L H₂O₂ and further increased during washout, causing irreversible contracture in one-half of the cells. The increase in [Ca²⁺]_i with 10 μmol/L H₂O₂ was modest with few cells showing irreversible contracture and attenuated by caffeine, and [Ca²⁺]_i gradually decreased during washout and this decrease was accelerated by a calcium-free solution, while 1 μmol/L H₂O₂ did not have any effects on [Ca²⁺]_i or cell viability. Ca²⁺ overload caused during exposure to 100 μmol/L H₂O₂ was attenuated by caffeine with improved cellular viability but not by chelerythrine, KB-R7943 or nifedipine. With 100 μmol/L H₂O₂ calcium-free solution attenuated the increase during exposure and washout while KB-R7943 or chelerythrine partly attenuated further increase during washout but not improved cell viability, but chelerythrine did not have additional effect on calcium-free treatment. Catalase abolished the effects of H₂O₂. We concluded that the increased [Ca²⁺]_i during exposure to 100 μmol/L H₂O₂ was caused both by release of Ca²⁺ from the intracellular store sites including the sarcoplasmic reticulum and by influx through route(s) other than the voltage-dependent Ca²⁺ channels or Na⁺/Ca²⁺ exchanger, although the Na⁺/Ca²⁺ exchanger or protein kinase C-mediated mechanism was partly responsible for a further increase during washout. Received: 1 February 2001, Returned for revision: 19 February 2001, Revision received: 11 April 2001, Accepted: 11 April 2001     

氧自由基在一氧化氮缺乏所致高血压及心血管重构中的作用

目的 :通过观察抗氧化剂Ebselen的干预效果 ,探讨氧自由基 (OFR)在一氧化氮 (NO)缺乏所致高血压、心血管重构中的作用。方法 :将 2 4只Wistar大鼠随机分为 3组 ,每组 8只 :①C组 :正常对照组 ;②L组 :L NAME 5 0mg·kg-1·d-1;③L +E组 :L NAME 5 0mg·kg-1·d-1加Ebselen 30mg·kg-1·d-1。隔周测动脉收缩压 ;8周末处死大鼠采集标本 ,检测血浆、心尖部心肌匀浆组织中NO含量及心肌超氧阴离子 (O-2 )水平、血管紧张素Ⅱ一型受体 (AT1受体 )表达 ;取心脏作病理形态检测。结果 :①L组血压逐渐升高 :自 1周末起 ,对比C组同期血压差异均有统计学意义 (P <0 .0 5 )。Ebselen干预早期对血压升高无明显影响 ,但在 8周末则明显低于L组 (P<0 .0 5 ) ;②L组心肌组织NO水平明显低于C组 ,L +E组则明显高于L组 ;③L组心肌O-2 、AT1受体表达水平 ,明显高于C组 ,L +E组则明显低于L组 ;④L组及L +E组心脏重量 /体重 (HW /BW )较对照组有显著增高 ;L +E组较L组有下降趋势 ,但P >0 .0 5。左室壁厚度L组及L +E组均较C组明显增高 (P <0 .0 5 ) ,而Ebselen干预后其左室壁厚度则明显变薄 (P <0 .0 5 ) ;心肌小动脉壁厚 /腔径比值的变化 :L组明显较对照组为大 ,L +E组与L组比较则此种比值有所减小 ,但L +E组仍较对照组为高 (P <0