大面积烧伤患者早期红细胞膜胆固醇变化与红细胞变形性的关系

目的研究大面积烧伤患者早期红细胞膜胆固醇与红细胞变形指数的变化及意义。方法利用核孔滤膜法测量红细胞变形指数,使用化学修饰电极测量红细胞膜胆固醇。结果大面积烧伤患者早期与正常对照组比较红细胞变形指数显著下降,分别为7.826±0.53及6.541±0.61,而红细胞膜胆固醇则显著升高,分别为(0.645±0.062)mmol/g膜蛋白及(0.891±0.051)mmol/g膜蛋白(P＜0.01),且二者呈高度负相关(r=-0.801)。结论红细胞膜胆固醇能够有效反映大面积烧伤患者早期红细胞变形能力,并成为反映大面积烧伤的早期微循环状态的新型量化指标。     

大面积烧伤患者早期红细胞膜胆固醇变化与红细胞变形性的关系

目的研究大面积烧伤患者早期红细胞膜胆固醇与红细胞变形指数的变化及意义。方法利用核孔滤膜法测量红细胞变形指数 ,使用化学修饰电极测量红细胞膜胆固醇。结果大面积烧伤患者早期与正常对照组比较红细胞变形指数显著下降 ,分别为 7.82 6± 0 .5 3及 6 .5 41± 0 .6 1,而红细胞膜胆固醇则显著升高 ,分别为(0 .6 45± 0 .0 6 2 ) mm ol/ g膜蛋白及 (0 .891± 0 .0 5 1) m mol/ g膜蛋白 (P<0 .0 1) ,且二者呈高度负相关 (r=- 0 .80 1)。结论红细胞膜胆固醇能够有效反映大面积烧伤患者早期红细胞变形能力 ,并成为反映大面积烧伤的早期微循环状态的新型量化指标     

严重烧伤对红细胞膜胆固醇含量及红细胞滤过指数的影响

目的　研究严重烧伤患者早期红细胞膜胆固醇与红细胞滤过指数的变化及意义。方法　利用核孔滤膜法测量红细胞滤过指数 ,使用化学修饰电极测量红细胞膜胆固醇。结果　严重烧伤患者早期与对照组比较红细胞滤过指数显著下降 ,而红细胞膜胆固醇则显著升高 (P <0 0 1) ,且二者呈高度负相关 (r =- 0 80 1)。结论　严重烧伤患者早期红细胞膜胆固醇含量的升高 ,有可能是引起红细胞滤过指数下降的一个重要原因。     

氧自由基血清镁离子变化在支气管哮喘中的作用与机制研究

目的　为了探讨哮喘患者体内氧自由基、镁离子与红细胞膜ATP酶的变化情况,以及它们与疾病发生程度和相关关系。方法　观察了40例哮喘发作期、30例缓解期者和健康对照组40例正常人,用生化方法测定了黄嘌呤氧化酶(XOD)、红细胞膜ATP酶、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性变化,过氧化脂质(LPO)、丙二醛(MDA)和血清镁钾、钠离子变化情况。结果　①哮喘发作组红细胞膜钙镁ATP酶、钠钾ATP酶活性明显低于正常对照组(P<001)。而钙镁ATP酶活性与LPO、MDA成高度负相关性,分别r=-0.5426、P<005,r=-06410、P<001。钠钾ATP酶活性与LPO、MDA也成显著负相关性,分别r=-0.4733、P<005,r=-04260、P<005。②哮喘发作组XOD、LPO、MDA均显著高于正常对照组,P<005和P<001。而SOD、CAT则显著低于正常对照组P<005。缓解组XOD比正常对照组也显著增高,但和发作期比较有显著下降P<005,SOD则有所提高但无统计学意义,CAT则比正常对照组显著降低,但比发作期有所提高,LPO与正常对照组比较有增高但无统计学意义,与发作期比较有显著降低P<005。③哮喘发作组血清镁离子与红细胞膜钙镁ATP酶、钠钾ATP酶活性均呈显著正相关r=03904、P<005,r=03797、P<005。结论　哮喘患者体内存在氧化与抗氧化失衡现象,氧自由基的损伤血清镁离子降低     

缺血性脑血管病患者血浆卵磷脂-胆固醇酰基转移酶活性与其脂质代谢

背景脂质代谢异常是缺血性脑血管病的危险因素之一.很多研究提出其与体内卵磷脂-胆固醇酰基转移酶活性变化有关.目的观察缺血性脑血管病患者血浆卵磷脂-胆固醇酰基转移酶活性与红细胞膜脂质成分含量变化的关系.设计病例对照(实验组对照,标准对照).单位一所大学医学院附属医院的检验科、急诊室及神经内科.对象2002-03/2003-12青岛大学医学院附属医院急诊神经门诊就诊及住院的脑血管病患者105例,均符合第二届全国脑血管病会议的诊断标准,选择脑动脉硬化患者42例和脑梗死63例构成两个患者组,其中男67例,女38例.同期选择在本院健康查体者65例构成对照组,男36例,女29例.方法采集参与者空腹血8 mL,采用酶学方法检测血浆卵磷脂-胆固醇酰基转移酶活性和血清高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、载脂蛋白A1和载脂蛋白B水平,采用邻苯二甲醛-醋酸硫酸方法测定红细胞膜胆固醇含量,采用化学定量法测定红细胞膜磷脂含量.主要观察指标患者组与对照组卵磷脂-胆固醇酰基转移酶活性变化及红细胞膜脂质成分含量的变化.结果按意向分析处理,105例患者组和65例对照组全部进入结果分析.①卵磷脂-胆固醇酰基转移酶活性脑动脉硬化组和脑梗死组活性变化均明显低于对照组[(2.14±0.72)kat/L,(2.06±0.80)kat/L,(2.61±0.74)kat/L,P＜0.01].②高密度脂蛋白胆固醇与载脂蛋白A1水平脑动脉梗化组与脑梗死组明显低于对照组[(11.32±0.33)mmol/L,(1.37±0.33)g/L,(1.28±0.33)mmol/L,(1.27±0.31)g/L,(1.60±0.43)mmol/L,(1.60±0.43)g/L,(t=2.72～5.01,P＜0.01)].③低密度脂蛋白胆固醇及红细胞胆固醇含量脑动脉硬化组与脑梗死组明显高于对照组[(2.94±0.82)mmol/L,(0.63±0.05)mmol/g,(3.02±0.79)mmol/L,(0.60±0.07)mmol/g,(2.56±0.58)mmol/L,(0.57±0.05)mmol/g,(P＜0.01)].并且卵磷脂-胆固醇酰基转移酶活性分别与高密度脂蛋白胆固醇及载脂蛋白A1呈正相关(r=0.247,P＜0.05;r=0.303,P＜0.01),而与低密度脂蛋白胆固醇和红细胞膜胆固醇呈负相关(r=-0.212,P＜0.05;r=-0.346,P＜0.01).结论缺血性脑血管病患者血浆卵磷脂-胆固醇酰基转移酶活性下降,且并非继发于脑梗死发生后,其活性变化与高密度脂蛋白胆固醇及载脂蛋白A1呈正相关,与低密度脂蛋白胆固醇及红细胞膜胆固醇呈负相关性.     

红细胞膜泵活性与血液粘滞性的关系及其对老年脑梗死的影响

目的探讨老年脑梗死(CI)病人红细胞膜Na＋—K＋ATP酶、Ca2＋—Mg2＋ATP酶与血液流变学指标的相关性。方法对49例非急性期的老年CI患者和29例健康人分别测定Na＋—K＋ATP酶、Ca2＋—Mg2＋ATP酶以及血液流变学指标。结果CI组全血粘度(ηb)、血浆粘度(ηp)、还原粘度(ηre)、红细胞压积(HCT)、红细胞聚集指数(EAI)、刚性(TK)与Na＋—K＋ATP酶、Ca2＋—Mg2＋ATP酶活力的变化呈明显负相关。结论CI患者红细胞膜泵活性的变化能引起血液粘滞性的改变,其作用主要是红细胞膜Na＋—K＋泵、Ca2＋泵的活性降低,导致红细胞变形能力下降。     

钠泵的基础与临床研究

钠泵与钠、钾-ATP酶钠泵(又称钠钾泵)是指细胞膜上具有主动转运钠钾离子功能的一种分子机构。在人和动物机体细胞膜内外,存在明显的电化学梯度,使钠钾离子分布具有很大的差异:细胞内呈高钾、低钠和负电势。这种差异须消耗机体的能量方得以维持。钠泵利用ATP水解释放的能量来对抗电化学梯度,实现钠钾离子的主动转运:泵出钠,泵入钾。1957年,Skou首先在一种蟹神经中发现了钠、钾依赖性三磷酸腺苷酶(钠、钾ATP酶,Na、K-ATPase,EC 3.6.1.3)。1965年,他证明了钠、钾-ATP酶具有与钠泵相同的性质:①位于     

临床水、电解质失调述要——二、钠代谢失调的临床(上)

【钠的代谢】细胞外液含钠量:血清为135～145毫当量/升,细胞间液则稍低;细胞内液的钠浓度约为10毫当量/升。由于两者之间相差10多倍,故钠不断由细胞外渗入细胞内,为了维持细胞内钠的衡定,钠泵要不断地将渗入的钠泵出,其能源是来自三磷酸腺苷(ATP),促进ATP的分解的ATP酶,其活力受细胞内液的钠钾浓度调节(钠、钾激活性ATP酶),是一种     

肾综合征出血热患者红细胞膜ATP酶活性改变的观察

目的探讨肾综合征出血热患者红细胞膜ATP酶活性的改变。方法按Reinila制膜法检测了33例肾综合征出血热患者红细胞膜Na ·K －ATP酶和Ca2 ·Mg2 －ATP酶含量。结果肾综合征出血热患者红细胞膜Na ·K －ATP酶活性低于正常人（P＜0．01）,而Ca2 ·Mg2 －ATP酶活性与正常人比较无差异（P＞0．05）。结论测定肾综合征出血热患者红细胞膜Na ·K －ATP酶和Ca2 ·Mg2 －ATP,酶活性的变化有助于临床的诊断和治疗。     

急性脑梗塞患者红细胞膜脂质过氧化改变和红细胞流变学的关系

本文对48例急性脑梗塞患者红细胞膜脂质过氧化物、血浆脂质过氧化物、红细胞滤过指数、红细胞电泳及血沉方程 K 值等项进行了测定,结果表明:病例组较正常对照组有显著性差异(P<0.01)。动态观察结果表明:急性脑梗塞病程1周内红细胞膜过氧化脂质水平最高,1月后逐渐下降接近正常。直线相关分析结果显示:红细胞膜脂质过氧化改变与红细胞聚集性、变形性有显著相关性。本文结果提示:急性脑梗塞患者细胞膜脂质过氧化改变可能是导致红细胞流变学异常的重要原因之一。     

Relation of erythrocyte Na+-K+ ATPase activity and cholesterol and oxidative stress in patients with type 2 diabetes mellitus

BACKGROUND: Diabetic patients are at high risk of atherosclerotic complications, and factors associated with this include hypercholesterolemia, hemorheologic disturbances in erythrocytes and oxidative stress. We, therefore, carried out a study in type 2 diabetic patients to determine the relationships of erythrocyte Na+-K+ ATPase activity, plasma cholesterol and oxidative stress in this population. METHODS: Erythrocyte Na+-K+ ATPase activity and its relationship between plasma cholesterol and thiobarbituric acid reactive substance (TBARS, a marker of oxidative stress) were studied in type 2 diabetic patients with (n = 26) or without angiopathy (n = 30). Na+-K+ ATPase activity was measured by a colorimetric enzymatic method. Plasma TBARS levels were determined spectrophotometrically. Diabetic patients were classified according to plasma cholesterol concentrations as normo- or hypercholesterolemic (plasma total cholesterol > 5.18 mmol/L). RESULTS: Diabetic patients with or without angiopathy had lower erythrocyte Na+-K+ ATPase activity (p < 0.001 and p < 0.001 respectively) and higher plasma TBARS levels than healthy subjects (n = 20) (p < 0.001 and p < 0.001 respectively). Na+-K+ ATPase activity in the diabetic patients with angiopathy was lower than in the diabetic patients without angiopathy (p < 0.001). In the diabetic patients both with and without angiopathy, hypercholesterolemic patients had lower erythrocyte Na+-K+ ATPase activity and higher plasma TBARS levels than normocholesterolemic patients (p < 0.001, p < 0.001 respectively) There was no difference in the plasma TBARS concentrations between diabetic patients with and without angiopathy. There were negative correlations between erythrocyte Na+-K+ ATPase activity and both plasma cholesterol (r = -0.72) and plasma TBARS (r = -0.46) and a positive correlation between plasma cholesterol and TBARS (r = 0.42). CONCLUSIONS: Elevated plasma cholesterol may be responsible for the inhibition of erythrocyte Na+-K+ ATPase activity. Together with elevated cholesterol, free radical-induced mechanisms may be involved in the inhibition of Na+-K+ ATPase activity.     

三七总皂苷对烫伤大鼠心肌细胞及红细胞膜ATP酶活性的影响

目的研究三七总皂苷(PNS)对烫伤早期心肌细胞膜(Ca2+-Mg2+)-ATP酶和(Na+-K+)-ATP酶活性的影响.方法采用30%体表面积Ⅲ度烫伤大鼠模型,用生化反应法分别测定心肌细胞膜与红细胞膜上(Ca2+-Mg2+)-ATP酶和(Na+-K+)-ATP酶活性变化.结果大鼠烫伤后心率加快,心电图T波低平,心肌细胞与红细胞膜(Ca2+-Mg2+)-ATP酶和(Na+-K+)-ATP酶活性均显著下降(P＜0.01),(Ca2+-Mg2+)-ATP酶活性在心肌细胞与红细胞之间呈显著正相关(r=0.951,P＜0.01).100mg/kg的PNS可使心率及T波恢复、ATP酶活性明显增加(P＜0.01或0.05).结论PNS显著增加烫伤大鼠心肌细胞膜(Ca2+-Mg2+)-ATP酶和(Na+-K+)-ATP酶活性是其改善心功能的重要机制之一.     

休克肠淋巴液对大鼠红细胞膜泵活性及氧自由基的影响

目的 观察休克肠淋巴液引流对失血性休克大鼠、休克肠淋巴液回输对正常大鼠红细胞膜泵活性及氧自由基的影响.方法 雄性Wistar大鼠24只,按随机数字表法分为假休克组、休克组、休克肠淋巴液引流组、休克肠淋巴液回输组,每组6只.休克组与引流组于麻醉和手术后复制失血性休克模型,引流组于休克1 h引流肠淋巴液;假休克组仅麻醉与手术;回输组于麻醉和手术后将引流的休克肠淋巴液回输.各组于休克3 h或相应时间点取腹主动脉血制备红细胞膜悬液,检测红细胞膜泵活性与自由基指标.结果 与假休克组比较,休克组红细胞膜Na+-K+-ATP酶(μmol·mg-1·h-1)、Ca2+-ATP酶(μmol·mg-1·h-1)、超氧化物歧化酶(SOD)活性(NU/mg)均显著降低(Na+-K+-ATP酶:0.039±0.011比0.068±0.010;Ca2+-ATP酶:0.035±0.016比0.087±0.015; SOD活性:0.785±0.289比1.202±0.328,P＜0.05或P＜0.01),丙二醛(MDA)含量(nmol/mg)显著升高(1.914±0.225比0.913±0.138,P＜0.01);与休克组比较,引流组红细胞膜Na+-K+-ATP酶活性(0.056±0.009)、Ca2+-ATP酶活性(0.079±0.025)、SOD活性(1.220+0.380)均显著升高,MDA含量(1.214±0.127)显著下降(P＜0.05或P＜0.01).与假休克组比较,回输组红细胞膜Na+-K+-ATP酶活性(0.050+0.013)、Ca2+-ATP酶活性(0.056±0.023)显著降低(均P＜0.05),SOD活性(0.862±0.288)有所降低(P＞0.05),MDA含量(1.456±0.270)显著升高(P＜0.01).结论 休克肠淋巴液回流是引起红细胞膜泵活性降低、加重红细胞膜自由基损伤的关键因素.     

Functionally Abnormal Na+-K+ Pump in Erythrocytes of a Morbidly Obese Patient

The Na(+)-K(+) pump in the erythrocytes of a mordibly obese patient shows a unique constellation of functional abnormalities. The number of pump units, measured by [(3)H]ouabain binding to intact cells, as well as the enzymatic activity of the (Na(+)-K(+))-dependent ATPase in erythrocyte membranes were found to be markedly increased compared with control cells (18-fold and 14-fold, respectively). There was a concomitant fivefold increase in the rate of pump-mediated uptake of (86)Rubidium (a K analogue); this was balanced by an increased rate of (86)Rb efflux. In striking contrast to normal cells, however, a major portion of this efflux (80%) was inhibited by ouabain, and thus appeared to be mediated by the Na(+)-K(+) pump.Erythrocytes from this patient had elevated levels of intracellular K(+) and reduced levels of intracellular Na(+). This finding, taken together with the ouabain inhibition of K(+) efflux and the absence of associated abnormalities, argues against the possibility that the increased number of Na(+)-K(+) pump units was a compensation for a primary increase in the permeability of the erythrocyte membrane to monovalent cations, as is seen in a variety of erythrocyte disorders. Further evidence for a primary abnormality of the enzyme was our observation that the cardiac glycoside ouabain bound to these cells with reduced affinity and had a right shifted dose response for pump inhibition. The markedly increased number of Na(+)-K(+) pump units in these cells did not appear to extend to mononuclear leukocytes.In conclusion, the erythrocytes from this patient have a very large number of functionally abnormal Na(+)-K(+) ATPase units. A unique abnormality of the erythrocyte Na(+)-K(+) ATPase of these cells is the most likely explanation for these findings.     

Decreased activity of the red blood cell ATPase-dependent Na+ pump in patients with cardiac syndrome X.

Marked Na(+)/Li(+) countertransport hyperactivity and post-load hyperinsulinaemia have been described in 93% of patients with cardiac syndrome X. We hypothesized that more complex abnormalities in Na(+) traffic across the cell membrane are present in these patients. The aim of the present study was to evaluate the activities of the two main transporters responsible for transmembrane Na(+) transport, i.e. the ATPase-dependent Na(+) pump and the Na(+)-K(+)-2Cl(-) co-transporter, in a selected group of patients with cardiac syndrome X. We evaluated 19 patients with cardiac syndrome X and 14 control subjects. The ATPase-dependent Na(+) pump and Na(+)-K(+)-2Cl(-) co-transport activities were assessed from Na(+)-loaded red blood cells by using nystatine, in the presence of furosemide and ouabain, as appropriate. Erythrocyte Na(+)/Li(+) countertransport activity, serum lipid and post-load (75 g of oral glucose) insulin levels were also evaluated. The V(max) of Na(+)/Li(+) countertransport (P=0.0001) and post-load insulin levels (120 min; P=0.001) were confirmed to be higher in patients with syndrome X than in controls. The V(max) of Na(+)-K(+)-2Cl(-) co-transport was similar in patients and controls. By contrast, the V(max) of the ATPase-dependent Na(+) pump was significantly lower (P=0.002) in syndrome X patients (3.13+/-0.87 mmol.h(-1).l(-1)) than in controls (4.28+/-1.10 mmol.h(-1).l(-1)). Serum total cholesterol and triacylglycerol concentrations were also higher in patients with syndrome X than in control subjects (P<0.0001). Thus decreased activity of the ATPase-dependent Na(+) pump was present in patients with cardiac syndrome X. Such an abnormality has the biological potential to augment microvascular tone and the response to constrictor stimuli via increased intracellular free Ca(2+). Of note, syndrome X patients also manifested Na(+)/Li(+) countertransport hyperactivity which, in turn, is known to induce peripheral insulin resistance and consequent abnormalities in insulin secretion and lipid turnover. Thus cardiac syndrome X appears as a multifaceted syndrome presenting with either metabolic or cardiovascular symptoms, or both, based on the expression of complex abnormalities in Na(+) traffic across the cell membrane.     

去除神经支配大鼠骨骼肌中钠钾三磷酸腺苷酶表达的变化

背景胰岛素诱导的离子运输改变可影响代谢调节,并且胰岛素抵抗条件下钠钾三磷酸腺苷酶活性的降低导致高血压以及糖原合成、葡萄糖氧化和ATP产生的减少.目的观察钠钾三磷酸腺苷酶在胰岛素抵抗模型中的表达调控.设计对照实验.单位英国邓迪大学生命科学学院分子生理学系.材料实验于1999-10/2000-02完成.使用15只体质量200～250 g的清洁级成年雄性Sprague Dawley大鼠.每次实验使用5只,相同实验重复3次.方法去除大鼠一侧后肢神经支配,4 d后将骨骼肌从去除神经支配的后肢和对侧对照后肢中解剖并分离,从红色肌肉(比目鱼与红色腓肠肌)和白色腓肠肌中分离粗制细胞膜.应用免疫印迹法和Northern杂交分析法分别检测蛋白质和mRNA的表达.使用Bio-Rad GS-670图像密度仪对蛋白质和mRNA的信号强度进行定量.主要观察指标对照侧和去除神经支配后肢骨骼肌中钠钾三磷酸腺苷各亚基蛋白质和mRNA表达水平的比较.结果15只大鼠全部进入结果分析.与对照侧相比,去除神经支配的后肢中①红色骨骼肌中α2和β1亚基的蛋白质表达分别下降46%和77%.在红色和白色骨骼肌中,α1亚基的蛋白质表达分别增加了20%和15%.β2亚基的蛋白质表达在红色和白色骨骼肌中均未发生显著变化.②在红色腓肠肌和比目鱼肌中,α2亚基的mRNA水平分别下降了29%和39%,β1亚基的mRNA水平分别下降了80%和52%.在红色和白色腓肠肌中,α1亚基的mRNA水平分别增加了9倍和1.3倍.β2亚基的mRNA水平在红色腓肠肌中无显著变化,但在白色腓肠肌中减少了59%.结论①在去除神经支配的大鼠骨骼肌中,钠钾三磷酸腺苷酶各亚基的表达受转录和转录后双重调控.②α2和β1亚基的蛋白质表达显著下降的结果提示,在大鼠骨骼肌中,去除神经支配造成的胰岛素抵抗使原本受胰岛素调节的钠钾三磷酸腺苷酶亚基的表达机制被破坏.     

中药补肾方对心力衰竭大鼠Na+-K+ATP酶、Ca2+-Mg2+ATP酶和琥珀酸脱氢酶的作用研究

目的研究补肾方对心力衰竭大鼠Na+-K+ATP酶、Ca2+-Mg2+ATP酶和琥珀酸脱氢酶(SDH)的作用。方法 60只大鼠随机分为模型组、假手术组、曲美他嗪组、补肾低、中、高剂量组,每组10只。采用结扎冠状动脉前降支法制备心力衰竭大鼠模型,术后2周开始灌胃,分别给予药物干预8周。8周后通过颈动脉插管记录大鼠血流动力学改变,包括左室收缩压(LVSP)、左室舒张压(LVEDP)、左室内压上升下降最大速率(±LVdp/dtmax);采用分光光度计检测心力衰竭大鼠心肌Na+-K+ATP酶、Ca2+-Mg2+ATP酶以及SDH水平。结果模型组大鼠心肌LVSP和+LVdp/dtmax较假手术组明显降低(P<0.01),而LVEDP和-LVdp/dtmax明显升高(P<0.05);补肾方干预后,中、高剂量大鼠心肌收缩、舒张功能明显优于模型组(P<0.01),以高剂量补肾方改善心力衰竭大鼠心功能明显。补肾方干预后,Na+-K+ATP酶、Ca2+-Mg2+ATP酶和SDH活力高于模型组,但低于假手术组,且随补肾方剂量增加,Na+-K+ATP酶、Ca2+-Mg2+ATP酶和SDH活力逐渐增强。结论补肾方可改善心力衰竭大鼠的心功能,可能与Na+-K+ATP酶、Ca2+-Mg2+ATP酶以及SDH活力增强相关。