醋氨酚大鼠肝细胞损伤及其机制的研究

醋氨酚（ＡＡＰ）引起大鼠肝细胞膜损伤发生在肝细胞还原型谷胱甘肽（ＧＳＨ）明显下降之后，提示ＧＳＨ下降是ＡＡＰ损伤肝细胞的原因之一。同步测定培养大鼠肝细胞ＧＳＨ和胞浆自由钙离子浓度（［Ｃａ＾２＋ｆ］ｃ）的结果证明：加１０ｍｍｏｌ／Ｌ ＡＡＰ培养２．５小时可引起ＧＳＨ显著降低，［Ｃａ＾２＋ｆ］ｃ明显升高。二巯基苏糖醇（ＤＴＴ）５ｍｍｏｌ／Ｌ可使加ＡＡＰ后，细胞ＧＳＨ仍维持较高水平，［Ｃａ＾２＋ｆ］ｃ明     

红细胞膜Ca^2＋—Mg^2＋—ATPase活性与慢性胃炎中医证型关系

从细胞和细胞膜酶分子水平探讨慢性胃炎中医证型病理生理特点,测定了３０例正常人和１０２例慢性胃炎病人基础和胰岛素刺激后红细胞膜Ｃａ＾２＋－Ｍｇ＾２＋－ＡＴＰａｓｅ活性,红细胞ＡＴＰ含量及２４ｈ尿１７－ＯＨＣＳ排出量。结果表明：湿热证膜Ｃａ６２＋－Ｍｇ＾２＋－ＡＴＰａｓｅ活性和细胞内ＡＴＰ含量呈代偿性增高;脾胃气虚酶活性和ＡＴＰ含量有所下降;脾肾气虚组织细胞代谢完全失代偿。     

Fas抗原在超抗原诱导T细胞凋亡中的作用

目的：研究Ｆａｓ抗原介导的死亡效应在超抗原诱导Ｔ细胞凋亡中的作用。方法：ＳＥＡ诱导从人外周血建立的短期ＳＥＡ反应Ｔ细胞系凋亡,１．８％琼脂凝胶ＤＮＡ电泳观察凋亡的特征条带;ＦＣＭ检测Ｆａｓ,ＦａｓＬ的表达量变化,Ｆｕｒａ－２／ＡＭ荧光批示剂检测胞浆「Ｃａ＾２＋」ｉ的变化。     

非胰岛素依赖型糖尿病患者红细胞膜ATPase活性改变及其影响因素

目的探讨非胰岛素依赖型糖尿病（ＮＩＤＤＭ）患者红细胞膜Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ及Ｃａ２＋－ＡＴ－Ｐａｓｅ活性改变的影响因素。方法测定７７例ＮＩＤＤＭ患者和５０例正常人血糖、总胆固醇（ＴＣ）、甘油三酯（ＴＧ）、高密度脂蛋白胆固醇（ＨＤＬＣ）、脂质过氧化物（ＬＰＯ）、谷胱甘肽（ＧＳＨ）、糖化血红蛋白（ＨｂＡｌｃ）、超氧化物歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨＰＸ）、红细胞膜Ｎａ＋－Ｋ＋－ＡＴ－Ｐａｓｅ和Ｃａ２＋－ＡＴＰａｓｅ活性。ｔ检验、直线相关分析和多元逐步回归分析。结果ＮＩＤＤＭ组血糖、ＨｂＡｌｃ、ＴＣ、ＴＧ、ＴＣ／ＨＤＬＣ、ＬＰＯ显著高于对照组（Ｐ＜０．０１），ＨＤＬＣ、ＧＳＨ、ＳＯＤ、ＧＳＨＰＸ、红细胞膜Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ和Ｃａ２＋－ＡＴＰａｓｅ活性低于对照组（除ＧＳＨ的Ｐ＜０．０５外，其他Ｐ＜０．０１）；不同ＨｂＡｌｃ、ＴＣ／ＨＤＬＣ、ＧＳＨ、ＬＰＯ、ＴＣ组的红细胞膜Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ活性有显著性差别。红细胞膜Ｎａ＋－Ｋ＋－ＡＴＰａｓｅ活性＝１２．８０＋０．２７（ＧＳＨ）－０．１２（ＬＰＯ）－０．０９（ＴＣ／ＨＤＬＣ），红细胞胞膜Ｃａ２＋－ＡＴＰａｓｅ活性＝１０８．２０＋１．     

利用培养乳鼠心肌细胞建立缺血再灌注损伤实验模型的探讨

利用培养乳鼠心肌细胞，复制缺血再灌注损伤模型，通过观察细胞三磷酸腺苷（ＡＴＰ）、细胞内钙离子（［Ｃａ＾２＋］ｉ）、脂质过氧化物（ＬＰＯ）、乳酸脱氢酶（ＬＤＨ）及心肌细胞扫描电镜变化，发现：①缺血３ｈ培养液中ＬＤＨ及［Ｃａ＾２＋］ｉ、ＬＰＯ轻度上升，ＡＴＰ耗竭明显，细胞严重水肿；②再灌注早期ＬＤＨ、［Ｃａ＾２＋］ｉ及ＬＰＯ急剧增加，ＡＴＰ含量逐渐升高，再灌注１ｈ细胞水肿减轻。结果提示：①在常温、缺氧     

非胰岛素依赖型糖尿病患者红细胞膜ATPase活性改变及其影 …

目的 搪塞非胰岛素依赖型糖尿病（ＮＩＤＤＭ）患者红细胞膜Ｎａ－Ｋ－ＡＴＰａｓｅ及Ｃａ＾２＋－ＡＴＰａｓｅ活性改变的影响因素。方法 测定７７例ＮＩＤＤＭ患者和５０例正常人血糖、总胆固醇（ＴＣ）、甘油三酯（ＴＧ）高密度脂蛋白胆固醇（ＨＤＬＣ）脂质过氧化物（ＬＰＯ）、谷胱甘肽（ＧＳＨ）、糖化血红蛋白、超氧化物歧化酶ＳＯＤ、谷胱甘肽过氧化物酶（ＧＳＨＰＸ）、红细胞膜Ｎａ－Ｋ－ＡＴＰａｓｅ和Ｃａ＾２＋－ＡＴ     

高钾离子引起PC12细胞内游离Ca2+浓度升高的机制

目的：分析高钾离子（Ｋ＾＋）引起ＰＣ１２细胞内游离钙离子浓度（［Ｃａ＾２＋］ｉ）升高的可能机制。方法：利用ＭｉｒａＣａｌ Ｉｍｉａｇｅ Ｓｙｓｔｅｍ检测［Ｃａ＾２＋］ｉ，Ｃａ＾２＋荧光探针为Ｆｕｒａ－２／ＡＭ。结果：（１）ＫＣ１浓度为３０￣１００ｍｍｏｌ／Ｌ时，可剂量依赖性地诱导ＰＣ１２细胞［Ｃａ＾２＋］ｉ升高；（２）在细胞外无Ｃａ＾２＋时，高Ｋ＾＋对ＰＣ１２细胞［Ｃａ＾２＋］ｉ无影响；（３）Ｌ－     

烫伤早期大鼠肾皮质细胞氧自由基,Ca^2＋细胞化学和Ca^2＋—Mg…

研究重度烫伤大鼠肾皮质细胞膜Ｃａ＾２＋－Ｍｇ＾２＋－ＡＴＰ酶活性，细胞内Ｃａ＾２＋定位和半定量及肾皮质内Ｈ２Ｏ２定位，以探讨烫伤早期肾损伤的机制。     

慢性肺心病急性期患者红细胞膜Na^＋—K^＋—ATPase,Ca^2＋—Mg^2…

应用酶学比色法测定３５例肺心病急性期患者及３０例健康人红细胞膜Ｎａ＾＋－Ｋ＾＋－ＡＴＰａｓｅ，Ｃａ＾２＋－Ｍｇ＾２＋－ＡＴＰａｓｅ活性，同时测定患者动脉血氧分压，动脉血二氧化碳分压。结果：患者红细胞膜Ｎａ＾＋－Ｋ＾＋－ＡＴＰａｓｅ，Ｃａ＾２＋－Ｍｇ＾２＋－ＡＴＰａｓｅ活性下降与对照组差异十分显著。     

凋亡相关基因bcl—2,bax,Fas及FasL在乙型肝炎病毒相关性？…

目的 探讨原发性肝癌中乙型肝炎病毒感染与凋亡基因ｂｃｌ－２、ｂａｘ、Ｆａｓ及ＦａｓＬ表达的关系。方法 采用免疫组化技术分析４０例原发肝细胞癌组织中乙肝病毒表现抗原（ＨＢｓＡｇ）阳性组和ＨＢｓＡｇ阴性组ｂｃｌ－２、ｂａｘ、Ｆａｓ及ＦａｓＬ的表达。结果 癌组织中ＨＢｓＡｇ阳性率为３０％（１２／４０）,癌旁组织中ＨＢｓＡｇ阳性率为６２．５％（２５／４０）,１２例ＨＢｓＡｇ阳性的癌组织中ｂｃｌ－２的ｂａｘ     

Method for human peripheral blood eosinophil isolation for patch-clamp study]

OBJECTIVE: To establish a rapid and economic method for isolating human peripheral blood eosinophils with high viability for patch-clamp studies and investigate the electrophysiological properties of Ca(2+)-activated K(+) channel of the isolated cells. METHODS: Peripheral blood eosinophils were isolated by modified discontinuous Percoll density gradient centrifugation, and the electric currents in the single Ca(2+)-activated K(+) channels of the cells were recorded using patch-clamp technique with cell-attached configuration. RESULTS: The purity of the eosinophils from healthy donors reached (90.5+/-1.6)%, with a viability rate over 99% and recovery rate of (48.2+/-6.9)%. The isolated cells were morphologically intact, from which Ca(2+)-activated K(+) channel activity could be detected. CONCLUSION: The peripheral blood eosinophils isolated using this rapid, simple and highly efficient method are characterized by high purity and viability without obvious cellular injuries, which are ideal for patch-clamp studies.     

活性氧对神经母细胞株SH-SY5Y细胞L型钙通道电流的影响

①目的 探讨氧自由基导致神经元钙超载的发生机制。②方法 在神经母细胞SH—SY5Y细胞体外培养的基础上，采用全细胞膜片钳技术，观察H2O2对SH—SY5Y细胞L型钙通道电流(ICa,L)的影响。③结果 H2O2能够明显增加峰值ICa,L，且该作用具有浓度依赖性。H2O2使ICa,L的I-V相关曲线下移，但对其形状和最大激活电位无明显影响。④结论 H2O2对L型钙通道有明显增强作用，此可能是其导致神经元钙超载的机制之一。     

细胞内Ca2+变化在砷剂诱导胰腺癌细胞株凋亡中的作用

目的：探讨细胞内游离Ca^2 在As2O3诱导的胰腺癌细胞株凋亡过程中的作用及Fas,Fas配体（FasL)的变化和意义。方法：2μmol/L的As2O3与胰腺癌细胞株SW－1990共同孵育后,H－E染色、光镜观察,抽提细胞DNA进行琼脂糖凝胶电泳,上流式细胞仪检测细胞凋亡特征; 用Fura-2荧光负荷技术测细胞内游离Ca^2 浓度（[Ca^2 ]i),流式细胞仪检测Fas,FasL的表达情况。结果：2μmol/LAsO3作用于胰腺癌细胞48h后,胰腺癌细胞出现典型的凋亡特性改变,且凋亡过程中[Ca^2 ]i明显升高,Fas,FasL表达呈先升后降,结论：As2O3在诱导胰腺细胞凋亡过程中,肿瘤细胞凋亡与[Ca^2 ]i升高和Fas、FasL表达有关。     

氯胺酮对谷氨酸诱导大鼠脊髓背角星形胶质细胞内游离钙浓度的影响

目的:观察氯胺酮对谷氨酸诱导大鼠脊髓背角星形胶质细胞凋亡和胞内游离钙浓度([Ca2+]i)变化的影响。方法:取新生2~3 d W istar大鼠T11~L6脊髓背角星形胶质细胞,原代纯化培养。将细胞随机分为:对照组(C组),谷氨酸组(G组),氯胺酮组(K组),余下3组为谷氨酸+不同浓度氯胺酮组(标记为GK1~GK3组),培养30 m in后取各细胞检测[Ca2+]i,再培养48 h检测星形胶质细胞凋亡率。结果:与C组比较,G组细胞发生了大量凋亡(P<0.01),[Ca2+]i显著升高(P<0.01);与G组比较,GK2组细胞凋亡率和[Ca2+]i明显降低(P<0.05),GK3组细胞凋亡率和[Ca2+]i显著降低(P<0.01)。结论:适量氯胺酮通过抑制细胞内钙超载显著抑制了谷氨酸诱导星形胶质细胞凋亡。     

EFFECTS OF THE HOE 694 ON TRANSIENTINWARD CURRENT AND NA＋－ CA2＋ EXCHANGE IN GUINEA PIG CARDIOMYOCYTES

OBJECTIVE: To determine the effects of HOE 694, a new and potent Na(+)-H+ exchanger blocker, on transient inward current (It(i)) and Na(+) - Ca(2+) exchange during hypoxia-reoxygenation in guinea pig cardiomyocytes. METHODS. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. RESULTS: In the absence of HOE 694, hypoxia-reoxygenation induced It(i) in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10 approximately 50 micro mol/L), the incidence of It(i) observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P < 0.05 vs control respectively. The Na(+) - Ca(2+) exchange current was unaffected by HOE 694 under normoxic condition. However, when cells were pretreated with 10 micromol/L HOE 694 for 10 min, then subjected to hypoxia condition, the Na(+) - Ca(2+) exchange current significantly inhibited. CONCLUSIONS: Blockade of the Na(+)-H+ exchange by HOE 694 could reduce Ca(2+) overload upon hypoxia-reoxygenation, and inhibition of Na(+) - H+ exchange may also indirectly decrease Na(+) - Ca(2+) exchange activity during hypoxia.     

Ionic mechanisms of a novel neurotoxin from the sea anemone Anthopleura sp. in rat ventricular myocytes.

OBJECTIVE: To determine the ionic mechanisms of a novel neurotoxin rhk2a obtained from the sea anemone Anthopleura sp. in rat ventricular myocytes. METHODS: Whole-cell patch-clamp recording technique was used to record the sodium, calcium, and sodium-calcium exchange currents (I(Na), I(Ca, L), and I(Na-Ca), respectively) in the isolated single rat ventricular myocytes with or without rhk2a treatment. RESULTS: The current-voltage (I-V) relationship for whole-cell I(Na) in the non-treated and rhk2a-treated (at the dose of 1 micromol/L) myocytes showed no significant difference (P>0.05), but the time constants for inactivation (tau(h)) were significantly greater (P<0.05) for the treated cells over the entire course of the experiment, while the time constants for activation (tau(m)) exhibited no significant difference between the two cells. The inactivation curve of I(Na) of rhk2a-treated cells was similar to that of the non-treated cells, as with the I-V relationship for whole-cell L-type calcium current (I(Ca, L) and I(Na-Ca)). CONCLUSIONS: Delayed inactivation of Na(+) channel plays an important role in the positive inotropic effect of rhk2a, possibly resulting from the alteration in Na(+) channel kinetics induced by rhk2a. rhk2a does not directly affect I(Ca, L), or I(Na-Ca).     

硫胺素和核黄素对H2O2诱导的DNA氧化损伤的影响

目的 探讨硫胺素和核黄素对H2O2诱导人脐静脉内皮细胞ECV304的DNA氧化损伤的影响.方法 于细胞培养液中分别加入硫胺素(终浓度为10、100、500、1000 mg/L)或核黄素(终浓度为20、100、300、500 nmol/L)预处理24 h,给予H2O2(终浓度为25 μmol/L)反应30 min,采用单细胞凝胶电泳技术(SCGE)分析DNA氧化损伤情况.以不加H2O2、硫胺素和核黄素者为阴性对照组;以加H2O2而不加硫胺素、核黄素者为阳性对照组.结果 H2O2诱导ECV304细胞DNA断裂损伤,SCGE分析显示,阳性对照组拖尾细胞率、彗星尾长、尾部DNA含量和Olive尾矩等指标均较阴性对照组升高;经10、100、500 mg/L硫胺素或20、100、300 nmol/L核黄素预处理后,各DNA损伤指标均较阳性对照组显著下降(P<0.05或P<0.01);经1000 mg/L硫胺素或500 nmol/L核黄素预处理后,各DNA损伤指标与阳性对照组比较差异无统计学意义(P>0.05).结论 适宜浓度的硫胺素和核黄素能降低H2O2诱导的DNA氧化损伤,高浓度的硫胺素和核黄素不能保护H2O2诱导的DNA氧化损伤.     

黄芪皂苷对过氧化氢诱导的肺泡上皮细胞凋亡的作用

目的探讨黄芪皂苷对过氧化氢(H2O2)引起的肺泡上皮细胞凋亡的拮抗作用。方法采用Annexin V评价细胞凋亡,Westernblot检测细胞内Fas和FasL,分别对过氧化氢单纯刺激组,黄芪100 uM和200 uM孵育后过氧化氢刺激组进行检测。结果黄芪皂苷孵育组凋亡数明显少于单纯H2O2刺激组(P<0.01)。黄芪皂苷孵育组中Fas和FasL的表达较单纯H2O2刺激组减弱(P<0.01)。结论黄芪皂苷可有效减少由H2O2引起的肺泡上皮细胞的凋亡,减少Fas和FasL的表达。     

胰腺腺泡细胞钙超负荷在诱发大鼠由水肿性向坏死性胰腺炎转变中 …

OBJECTIVE: To investigate the potential of pancreatic acinar cell calcium overload in the conversion of acute edematous pancreatitis (AEP) to necrotizing pancreatitis (ANP). METHODS: Ninety-six Sprague-Dawley rats were randomized in three experimental groups. Sham-operated control (Group I) AEP (Group II) was induced by pancreatic duct ligation and intravenous injection of bombesin (100 micrograms/kg) and secretin (10 micrograms/kg). ANP (Group III) was induced same as group II but with a large dose of dextran 110,000(500 mg/kg) intravenously. Pancreatic acinar cell Ca2+ overload was studied using fluorescent probe Fura2. Cytosolic free Ca2+ concentration ([Ca2+]i) in isolated pancreatic acinar cells and Ca(2+)-ATPase activity in pancreatic cell plasma membranes were determined 1, 3, 6, 9 h respectively after treatment. RESULTS: The results showed that pancreatic acinar cell [Ca2+]i was elevated at 1 h[(213 +/- 19) nmol/L, P < 0.05] and increased to (464 +/- 29) nmol/L at 6 h(P < 0.05) in rats with ANP, pancreatic cell plasma membrane Ca(2+)-ATPase activity decreased significantly from (29.8 +/- 0.4) nmol.min-1.mgp-1 at 1 h to (18.6 +/- 0.5) nmol.min-1.mgp-1 at 9 h in rats with ANP (P < 0.05). CONCLUSIONS: In ischemia-induced conversion of AEP to ANP, there exists Ca2+ overload in the pancreatic acinar cells. The decreased pancreatic cell plasma membrane Ca(2+)-ATPase activity may be an important reason for acinar cell Ca2+ overload in the development of acute pancreatitis.