特异性抗原诱发致敏豚鼠心室乳头肌早期后除极和触发活动的研究

采用微电极细胞内记录和电子计算机实时采样技术,研究了特异性抗原（卵白蛋白０．２５μｍｏｌ／Ｌ）对致敏豚鼠心室乳头肌动作电位的影响。特异性抗原激发后,致敏心肌动作电位发生了下列改变：２ｍｉｎ左右ＡＰＤ＿（５０）、ＡＰＤ＿（９０）明显延长;５ｍｉｎ后,ＡＰＤ＿（５０）、ＡＰＤ＿（９０）、３期时程明显缩短;２０ｍｉｎ以后,上述改变逐渐恢复。心性过敏反应可诱发早期后除极（发生率５５％）和触发活动,形成快速自发动作电位（发生率４０％）,早期后除极在低频驱动（０．２－１．０Ｈｚ）时易于发生,且在２ｍｉｎ左右多见。结果提示,心性过敏反应诱发的动作电位时程的变化、早期后除极和触发活动,可能和超敏反应时发生的快速自律型心律失常有关。     

组胺受体在致敏豚鼠心室肌延迟后除极和触发活动中的作用

采用微电极细胞内记录和电子计算机实时采样技术,研究了组胺受体在特异性抗原（卵清白蛋白,０．２５μｍｏｌ／Ｌ）对致敏豚鼠心室乳头肌动作电位的影响中的作用。心性过敏反应可诱发延迟后除极（发生率６５％）和触发活动,并形成快速自发动作电位。高频刺激时,易产生延迟后除极。外源性组胺（０．６μｍｏｌ／Ｌ和６．０μｍｏｌ／Ｌ）作用于豚鼠心室乳头肌亦产生相似的变化。Ｈ＿２受体阻断剂西咪替丁（１０μｍｏｌ／Ｌ,ｎ＝１０）可明显抑制心性过敏反应和完全阻断组胺所致的延迟后除极现象,Ｈ＿１受体阻断剂扑尔敏（１０μｍｏｌ／Ｌ,ｎ＝６）对心性过敏反应和组胺的效应均无抑制作用。结果提示。心性过敏反应诱发的心肌延迟后除极,可能是心肌释放的组胺直接作用于心室肌Ｈ＿２受体所致。     

特异性抗原诱发致敏豚鼠心室乳头肌早期后除极和触发活动的…

采用微电极细胞内记录和电子计算机实时采样技术,研究了特异性抗原对致敏豚鼠心室乳头肌动作电位的影响。特异性抗原激发后,致敏心肌动作电位发生了下列改变：２ｍｉｎ左右ＡＰＤ５０、ＡＰＤ９０明显延长;５ｍｉｎ后,ＡＰＤ５０、ＡＰＤ９０、３期时程明显缩短;２０ｍｉｎ以后,上述改变逐渐恢复。心性过敏反应可诱发早期后除极（发生率５５％）和触发活动,形成快速自发动作电位（发生率５０％）,早期后除极在低频驱动时易于     

组胺致豚鼠心室乳头肌早发后去极化和触发活动的研究

采用微电极细胞内记录和电子计算机实时采样技术,研究了组胺致豚鼠心室乳头肌早发后去极化和触发活动。组胺(6.0μmol/L)可使动作电位APD_(50)、APD_(90)明显缩短(数据取自无异常自律性和振荡电位的细胞)。组胺可诱发早发后去极化(发生率57％),振荡电位和触发活动。早发后电位在低频驱动(0.2～1.0Hz)时易于发生且常常伴有动作电位时程的延长。触发活动可表现为快速自发性动作电位(发生率14％)。结果提示,组胺诱发的早发后去极化和触发活动可能和心性过敏反应等疾病发生的快速自律型心律失常有关。     

丹皮酚对心肌细胞自律性和延迟后除极的影响

目的与方法：采用常规玻璃微电极技术研究丹皮酚对离体心肌细胞自律性（ＡＭ）、延迟后除极（ＤＡＤ） 及触发活动（ＴＡ）的影响。结果：１．８×１０－４ｍｏｌ／Ｌ丹皮酚灌流组,肾上腺素（Ａｄｒ）的阈浓度空白对照组为（１．２８±０．５７）μｍｏｌ／Ｌ,药后为（１．５６±０．５３）μｍｏｌ／Ｌ（ｎ＝９,Ｐ＞０．０５）;用（１．８×１０－ ３） ｍｏｌ／Ｌ丹皮酚（Ｐａｅ）灌流组,Ａｄｒ 浓度由空白对照组的（１．２２ ±０．６２）μｍｏｌ／Ｌ升高到（６．２２±２．１１）μｍｏｌ／Ｌ（ｎ＝９,Ｐ＜０．０１）。１．８×１０－３ｍｏｌ／Ｌ的Ｐａｅ 能明显抑制哇巴因（Ｏｕａ）诱发的ＤＡＤ的幅值,当基本刺激周长为５００,４００,３００ 和２００ ｍｓ 时,其ＤＡＤ幅值从（５．５±２．０）ｍＶ,（７．３±２．１）ｍＶ,（８．０ ±２．４）ｍＶ和（９．２±１．９）ｍＶ减小到（３．０±１．１）ｍＶ、（３．６±１．７）ｍＶ,（４．３±２．０） ｍＶ和（５．９ ±１．６） ｍＶ,Ｐ＜０．０１。当基本刺激周长为２００ ｍｓ时,ＴＡ 数目由５．５±１．０ 降至０．７±０．３（Ｐ＜０．０１）。结论：丹皮酚能抑制心肌细胞ＡＭ、ＤＡＤ及ＴＡ,具有抗心律失常作用     

心肌缺O2再给O2时延迟后除极和触发活动的发生及机制

应用细胞内微电极技术观察了豚鼠右心室乳头肌缺O_2/再给O_2与延迟后除极(DADs)及触发活动的关系;以及高Ca~(2 )、Ca~(2 )通道阻断剂、吗啡对再给O_2过程诱发的DADs及触发活动的效应。结果表明:单纯缺O_2和再给O_2期间均未产生DADs及触发活动,在缺O_2和葡萄糖条件下,再给O_2和葡萄糖时能诱发出DADs及触发活动,并且DADs幅度与缺O_2和葡萄糖的持续时间有关。高Ca~(2 )(由2.45mmol/L增加到4.9mmol/L)明显增加再给O_2和葡萄糖期间产生的DADs幅度和触发活动发生数;异搏定(1mg/L)明显减低再给O_2和葡萄糖期间产生的DADs幅度。吗啡(120μmol/L)明显减低再给O_2和葡萄糖期间产生的DADs幅度,且吗啡这种作用可被纳洛酮(5μmol/L)所阻断。这些实验表明:DADs和触发活动的发生与心肌能量状态、缺O_2时间的长短、细胞外液中Ca~(2 )浓度有密切关系。异搏定和吗啡可明显降低DADs及触发活动。     

大鼠心室肌对低温的耐受性及触发活动的产生

用标准微电极技术记录大鼠心室肌经低温处理1～3d后某些电学性质的变化。低温处理后的标本,复温到36±1℃后,动作电位幅度无明显改变,但动作电位时程(APD_(90))显著延长。快速驱动(10～30Hz)可使低温处理标本产生迟后去极化(DAD)和触发活动(TA)。并且驱动频率越高,持续时间越长,所引起的DAD幅度越大,越易发生TA。无K~ 灌流、哇巴因1～4×10~(-5)mol/L以及咖啡碱7.1～14.2×10~3mol/L处理可使低温处理标本产生DAD和TA。结果表明,大鼠心室肌细胞能耐受较长时间的低温处理。但经处理后,心室肌的某些电学性质发生改变,它们对能引起细胞内Na~ 或Ca~(2 )增加的因素更为敏感,更易产生DAD及TA。     

在体心脏后去极化及触发性心律失常的细胞电生理学研究

应用漂浮微电极记录跨膜动作电位(TAP)和接触电极记录单相动作电位(MAP)两项技术,研究了 CsCl 诱发的猫在体心脏的触发性活动。结果表明,静脉注射 CsCl(0.5mmol/kg)后10s,左心室外膜 TAP 和 MAP 的3相中晚期出现早期后去极化(EAD),在30s 时,TAP上 EAD 的振幅为25.6±9.3mV,MAP 上的 EAD 振幅为3.4±1.3mV。EAD 表现为拖尾、平台、凸起三种不同形态。在两例猫中出现延迟性后去极化 DAD,其在 TAP 和 MAP上的振幅分别为13.0±5.3mV 和3.3±0.omV。MAP 上后去极化的形态与 TAP 极为相似。CsCl 的重复注射可诱发室性早搏、室性心动过速等多种心律失常,根据后去极化的发生与否以及后去极化和室性心搏的耦联间期的关系,可将心律失常分为两种类型:一种是由记录部位的后去极化引起的触发性心律失常;另一类可能由非记录部位的后去极化或其它机制引起。     

氯胺酮诱导不同年龄和性别小鼠的过度活动

虽然氯胺酮(Ketamine)在历史上作为麻醉剂用于人类和家畜,但由于其分解特性, 作为一种娱乐药物似乎更具知名度.以前的研究表明相对于成年人, 孩子使用氯胺酮不易显示出不利影响,但是对其发生的机理几乎没有研究.本文研究了氯胺酮对小鼠的活动程度和固有行为的作用.结果表明:使用氯胺酮可增加22、35和50日龄小鼠的运动器官的敏捷性,并证明了氯胺酮的作用随年龄的增长而降低;使用氯胺酮所导致的旋转与年龄的变化有关,但站立的减少与年龄无关,这种减少不依赖于小鼠的年龄.     

七种Mg2+转运体在出生后不同时期小鼠心肌细胞的表达

目的:研究小鼠心肌细胞上七种Mg2+转运体mRNA在不同发育时期的表达情况.方法:提取不同发育时期小鼠心脏组织mRNA,用七对Mg2+转运体特异性引物进行RT-PCR扩增,经凝胶成像仪进行凝胶荧光定量检测.结果:经RT-PCR半定量检测后,观察到小鼠心肌细胞上七种转运体mRNA在不同时期都有不同程度的表达,其中TRPM7和MagT1两种转运蛋白在各个时期表达较稳定,并且表达量明显高于其他五种Mg2+转运体.结论:TRPM7和MagT1在小鼠心肌细胞不同发育时期Mg2+调控过程中可能发挥主导作用.     

HLAMatchmaker软件分析Eplets的配型在临床肾移植配型中的应用

目的：评估基于Eplets错配的HLAMatchmaker软件的方法在临床肾脏移植的应用。方法：针对239例肾移植供受者甩A—Ⅰ、Ⅱ抗原的血清学和基因型,比较传统的HLA六抗原配型标准、交叉反应组配型标准和基于HLA Matchmaker软件Eplets配型三种方法优劣关系。结果：239例供受者用三种方法得出错配数,按照高中低错配分组,结果显示HLAMatchmaker软件配型方法能明显增加低错配组别中的人数22．6％（54）,相比较于HIA配型的7．9％（19）和CREGs配型的13．4％（32）,三种方法之间有统计学意义（P〈0．01）;此外对三种方法所得的错配数两两比较也显示出统计学差异（P〈0．01）。结论：采用基于Eplets的HLAMachmaker软件配型进行供受者配型的方法,是一种更高效省时高灵敏度的同种异体肾移植的配型方案。     

植物性雌激素genistein对哇巴因所致豚鼠乳头状肌迟后去极化及触发活动的效应

应用标准玻璃微电极技术,研究了植物性雌激素genistein(GST)对哇巴因所引起的豚鼠乳头状肌迟后去极化（DAD）及触发活动（TA）的效应,结果如下：（1）预先给予是GST（10,50,100umol/L)剂量依赖性地抑制哇巴因（1umol/L)所引起的鼠乳头状肌DAD及TA;（2）预先应用一氧化氮合酶抑制剂L－NAME（1mmol/L),不影响GST（50μmol/L）对DAD及TA的效应;（3）单独应用17β－雌二醇（E2,5μmol/L）或GST（10μmol/L）对DAD及TA无明显影响,而联合应用相同剂量的GST和E2则产生明显儿应,以上结果提示,GST可能通过抑制钙离子内流从而具有抗心律失常作用,这对于心血管系统的保护有一定意义。     

Alterations in monoamine oxidase activity of the mouse brain and liver after mixed neutron-gamma irradiation

Monoamine oxidase (MAO) plays an important role in the metabolism of neuro-transmitter biogenic amines. Its activity was determined in mouse brain and liver after exposure to different kinds of ionizing radiation and after pretreatment with a radioprotective agent. After a lethal dose of mixed neutron-gamma irradiation the MAO activity decreased in the brain and increased in the liver. In contrast, after a lethal dose of 60Co-gamma irradiation enzyme activity was considerably increased in the brain while in the liver it increased like after mixed neutron-gamma irradiation. AET (S2-aminoethyl-isothiuronium-Br X HBr), when administered in a radio-protective dose, inhibited MAO activity in the brain, while it increased in the liver. Even more marked changes of enzyme activity were observed in both brain and liver after AET pretreatment and mixed neutron-gamma irradiation. On the basis of the results it is suggested that different kinds of ionizing radiation lead to different types of lipid peroxidation in the lipid environment surrounding MAO, an event leading to altered enzyme activity. AET itself inhibited MAO in the brain and increased the activity in the liver but did not prevent the alterations caused by ionizing radiation in enzyme activity.     

Alterations in protein kinase C isoenzyme expression and autophosphorylation during the progression of pressure overload-induced left ventricular hypertrophy

Cardiomyocytes express several isoenzymes of protein kinase C (PKC), which as a group have been implicated in the induction of left ventricular hypertrophy (LVH) and its transition to heart failure. Individual PKC isoenzymes also require transphosphorylation and autophosphorylation for enzymatic activity. To determine whether PKC isoenzyme expression and autophosphorylation are altered during LVH progression in vivo, suprarenal abdominal aortic coarctation was performed Sprague-Dawley rats. Quantitative Western blotting was performed on LV tissue 1, 8 and 24 weeks after aortic banding, using antibodies specific for total PKC, PKC and PKC, and their C-terminal autophosphorylation sites. Aortic banding produced sustained hypertension and gradually developing LVH that progressed to diastolic heart failure over time. PKC levels and autophosphorylation were not significantly different from sham-operated controls during any stage of LVH progression. PKC expression levels were also unaffected during the induction of LVH, but increased 3.2 ± 0.8 fold during the transition to heart failure. In addition, there was a high degree of correlation between PKC levels and the degree of LVH in 24 week banded animals. However, autophosphorylated PKC was not increased at any time point. In contrast, PKC autophosphorylation was increased prior to the development of LVH, and also during the transition to heart failure. The increased PKC autophosphorylation in 1 week banded rats was not accompanied by an increase in total PKC, whereas total PKC levels were markedly increased (6.0 ± 1.7 fold) in 24 week banded animals. Furthermore, both phosphorylated and total PKC levels were highly correlated with the degree of LVH in 24 week banded rats. In summary, we provide indirect evidence to indicate that PKC may be involved in the induction of pressure overload LVH, whereas both PKC and PKC may be involved in the transition to heart failure.     

Bone marrow derived cells and reactive oxygen species in hypertrophy of contralateral kidney of transient unilateral renal ischemia-induced mouse

Abstract

Renal mass reduction, such as unilateral nephrectomy induces a compensatory hypertrophy of remaining renal mass in response to overload induced by reduction of functional renal parenchyma. In our recent study, we observed that the recovery of ischemic injured kidney following transient unilateral renal ischemia took longer time than that following transient bilateral renal ischemia, indicating that non-damaged kidney may affect the damaged kidney and vice versa. Here, we investigated whether transient and partial renal parenchymal injury by transient unilateral renal ischemia (UI) results in the hypertrophy of its contralateral kidney (CLK) and reactive oxygen species is associated with the hypertrophy. Thirty minutes of UI resulted in gradual increase in CLK weight over time. UI increased superoxide formation, but not lipid peroxidation in the CLK. After UI, a significant increase in the number of NADPH oxidase 2 (Nox2)-expressing cells and the level of Nox2 expression in the CLK was observed. In parallel with the increases in Nox2-expressing cells in CLKs, infiltration of bone marrow-derived cells (BMDC) increased in CLK. Treatments with Mn(III) Tetrakis(1-methyl-4-pyridyl) porphyrin (MnTMPyP, a superoxide dismutase (SOD) mimetic) and apocynin (a putative NADPH oxidase inhibitor) inhibited UI-induced hypertrophy of CLK along with reduction in Nox2-positive cell, BMDC, amount of Nox2 expression and superoxide formation. In conclusion, transient and partial renal mass reduction by UI resulted in the hypertrophy of CLK through increased ROS formation by infiltrated cells into the interstitium of CLK.     

Alterations of mitochondrial protein metabolism in liver, brown adipose tissue and skeletal muscle during cold-acclimation

Incorporation of -[U-¹⁴C] leucine into liver, brown adipose tissue and skeletal muscle mitochondrial proteins was determined in vivo and in vitro during cold-acclimation. Major alterations in mitochondrail protein metabolism were observed in brown adipose tissue and skeletal muscle but not in liver. Immediate cold-exposure is accompanied by an inhibition of the in vivo incorporation of -[U-¹⁴C] leucine into mitochondrial proteins of all tissues. However, during cold-acclimation the incorporation of leucine increases markedly in brown adipose tissue, continues to decrease in skeletal muscle, nut does not change appreciably in the liver. Because increased incorporation of -[U-¹⁴C] leucine into brown adipose tissue mitochondrial proteins was observed both in vivo and in vitrom it can be concluded that the mitochondrial protein-synthesizing system of this tissue is directly affected by the acclimation process. The observed changes in mitochondrial protein metabolism of brown adipose tissue and skeletal muscle might be responsible for the development of several morphological and biochemical alterations that characterize the establishment in these tissues of the cold-acclimated state.     

Alterations in Ca2+-ATPase activity and calcium-binding protein regucalcin mRNA expression in the kidney cortex of rats with saline ingestion

The alteration in calcium metabolism in rats ingested with saline was investigated. Rats were freely given saline as drinking water for 2 and 7 days. Calcium concentration in the serum was significantly elevated by saline ingestion for 2 and 7 days, while serum inorganic phosphorus concentration was not altered. Serum urea nitrogen concentration was significantly increased by saline ingestion for 7 days. Calcium content in the femoral-diaphyseal and metaphyseal tissues was not altered by saline ingestion for 7 days. Calcium content in the kidney cortex was significantly elevated by saline ingestion for 7 days. Ca2+-ATPase activity in the basolsateral membranes of kidney cortex was clearly increased by saline ingestion for 2 and 7 days. The enzyme activity was not altered by the addition of sodium chloride (10-3 and 10-2 M), parathyroid hormone (10-7 and 10-6 M), and calcitonin (3 × 10-8 and 3 × 10-7 M) in the enzyme reaction mixture. A calcium-binding protein regucalcin mRNA expression in the kidney cortex was markedly suppressed by saline ingestion for 7 days, although such a suppression was not seen for 2 days. These results suggest that saline ingestion causes the disturbance of calcium transport system in the kidney cortex of rats, and that the renal disorder may induce hypercalcemia.     

Structural and functional roles of desmin in mouse skeletal muscle during passive deformation

Mechanical interactions between desmin and Z-disks, costameres, and nuclei were measured during passive deformation of single muscle cells. Image processing and continuum kinematics were used to quantify the structural connectivity among these structures. Analysis of both wild-type and desmin-null fibers revealed that the costamere protein talin colocalized with the Z-disk protein alpha-actinin, even at very high strains and stresses. These data indicate that desmin is not essential for mechanical coupling of the costamere complex and the sarcomere lattice. Within the sarcomere lattice, significant differences in myofibrillar connectivity were revealed between passively deformed wild-type and desmin-null fibers. Connectivity in wild-type fibers was significantly greater compared to desmin-null fibers, demonstrating a significant functional connection between myofibrils that requires desmin. Passive mechanical analysis revealed that desmin may be partially responsible for regulating fiber volume, and consequently, fiber mechanical properties. Kinematic analysis of alpha-actinin strain fields revealed that knockout fibers transmitted less shear strain compared to wild-type fibers and experienced a slight increase in fiber volume. Finally, linkage of desmin intermediate filaments to muscle nuclei was strongly suggested based on extensive loss of nuclei positioning in the absence of desmin during passive fiber loading.     

葡萄籽中原花青素对实验动物主动脉收缩和血小板聚集的影响

目的:研究葡萄籽中原花青素(PA)对大鼠离体主动脉平滑肌收缩活动和兔血小板聚集的影响.方法:采用大鼠离体主动脉环灌流方法,记录主动脉环张力变化,观察PA对去甲肾上腺素(NA)和KCl预收缩大鼠离体主动脉平滑肌收缩反应的舒张作用以及对NA量效曲线的影响.比浊法测定兔血小板聚集.结果:PA能明显抑制NA(10-6mol/L)预收缩大鼠离体主动脉环的反应,使NA量效曲线压低,最大反应降低,此作用无内皮依赖性,但对KCl预收缩主动脉环的舒张作用无明显影响,也不影响花生四烯酸(AA),ADP和胶原(collagen)蛋白诱导的兔血小板聚集.结论:PA能对抗NA而不影响KCl诱导的大鼠离体主动脉平滑肌的收缩,不影响兔血小板聚集.