“大通牦牛”胎盘-胎儿组织线粒体SDH、CCO和ATP酶活性的测定

为探究牦牛发育早期低氧适应的生物学机制,以"大通牦牛"(海拔3 200 m左右)为研究对象,采用酶联免疫分析法(ELISA)和比色法对妊娠期4月龄牦牛胎盘、胎儿及出生1日龄牦牛组织中琥珀酸脱氢酶(SDH)、细胞色素C氧化酶(CCO)及三磷酸腺苷(ATP)酶活性进行测定。结果:胎儿和犊牦牛骨骼肌、心肌、肝脏及大脑各组织线粒体SDH、CCO活性差异均不显著(P>0.05);妊娠4月龄胎盘组织和胎儿肝脏组织线粒体中Na+-K+-ATP、Mg2+-ATP、Ca2+-ATP和Ca2+-Mg2+-ATP 4种ATP酶活性均高于心肌、骨骼肌、大脑组织,差异显著(P<0.05);出生1日龄牦牛骨骼肌、心肌、肝脏及大脑各组织间4种ATP酶差异不显著(P>0.05),各组织中Ca2+-Mg2+-ATP酶活性均高于其他ATP酶活性。研究表明牦牛在发育早期组织线粒体中SDH、CCO保持相对稳定的水平,而ATP酶在物质运送、能量转换以及信息传递方面发挥重要的作用。     

高海拔地区牦牛心肌骨骼肌ATP酶活性的测定

为了探讨高原动物对高原低氧的适应机制,采用标准ATP酶测试盒对青海省(平均海拔4 000 m)16头成年牦牛心肌、骨骼肌ATP酶活性进行了测定和分析.结果:Na+ -K+ -ATP,Mg2+ -ATP,Ca2+ ATP和Ca2+-Mg2+-ATP 4种酶活性在骨骼肌线粒体中分别为3.65±1.31(μmolP/mg蛋白·h,下同),2.70士1.36,2.87±1.23和4.08士1.30;在心肌线粒体中分别为4.44±1.19,2.42士0.87,2.29±0.57和3.62±0.92;在骨骼肌组织中分别为2.02±0.80,1.54士0.36,1.69士0.91和2.02±0.96;在心肌组织中分别为1.41士0.67,1.04士0.51,1.24±0.54和1.42±0.60;骨骼肌、心肌线粒体中4种ATP酶活性均显著高于骨骼肌、心肌组织(P＜0.05).     

噻拉嗪对山羊不同脑区ATP酶活性的影响

为研究噻拉嗪对山羊不同脑区Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的影响,探讨噻拉嗪麻醉山羊的中枢作用机制。试验将25只山羊随机平均分成对照组、诱导组、麻醉组、恢复Ⅰ组和恢复Ⅱ组共5组。对照组肌肉注射生理盐水10 m L,试验组肌肉注射噻拉嗪12.8 mg/kg体重。10 min后、山羊倒地、翻正反射消失后、翻正反射恢复后和翻正反射恢复后6 h断头取脑组织,采用分光光度法测定山羊各脑区Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性。结果大脑皮质、丘脑的Na+-K+-ATP酶活性分别较对照组降低30.1%和21.9%(P<0.01);大脑皮质、海马、丘脑、小脑和脑干内Ca2+-Mg2+-ATP酶活性分别较对照组降低36.5%、38.6%、35.1%、36.9%和37.9%(P<0.01)。恢复Ⅰ组和恢复Ⅱ组两种ATP酶活性的变化与对照组相比差异均无统计学意义(P>0.05)。表明噻拉嗪通过大脑皮质和丘脑Na+-K+-ATP酶及大脑皮质、海马、丘脑、小脑和脑干内Ca2+-Mg2+-ATP酶活性作用的改变起麻醉作用。     

氯胺酮对大鼠海马神经元ATP酶活性的影响

本试验研究不同浓度氯胺酮对大鼠海马神经元ATP酶活性的影响,并探讨氯胺酮的中枢麻醉机制。体外培养怀孕16~18 d的胎鼠神经细胞,至第8天直接给予3种不同浓度的氯胺酮,使用超微量ATP酶测定试剂盒,检测胎鼠海马中ATP酶的活力。结果显示,海马中Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶的活性均受到不同程度的抑制,并且这种抑制作用的程度具有剂量依赖的趋势。据此推测氯胺酮的麻醉机制可能与抑制海马中Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶的活性有关。     

乳化异氟醚静脉麻醉对大鼠中枢突触体ATP酶活性的影响

为探讨乳化异氟醚对大鼠中枢突触体ATP酶活性的影响,选用36只Wistar大鼠随机分为对照组、麻醉组和麻醉恢复组,经乳化异氟醚尾静脉注射对大鼠进行麻醉,采用比色法检测各脑区的突触体ATP酶活性。结果显示:大鼠注射乳化异氟醚后大脑皮层、海马和丘脑Na+-K+-ATP酶和Ca2+-ATP酶与对照组相比显著下降(P0.05),大脑皮质和海马Mg2+-ATP酶与对照组相比显著下降(P0.05)。提示:乳化异氟醚能够抑制大脑皮层、海马和丘脑内Na+-K+-ATP酶和Ca2+-ATP酶的生成,抑制大脑皮质和海马内Mg2+-ATP酶的生成,推测中枢突触体ATP酶是乳化异氟醚产生麻醉效应的主要靶位。     

鹿特异性复合麻醉剂对大鼠不同脑区ATP酶活性的影响

为研究鹿特异性复合麻醉剂麻醉与大鼠各脑区突触体ATP酶活性的关系,探讨其麻醉机理。将20只SD大鼠分为对照组和麻醉组,对照组大鼠腹腔注射30mL/kg生理盐水,试验组腹腔注射30mg/kg鹿特异性复合麻醉剂,采集大鼠各脑区,利用比色法测定脑区内Na＋-K＋-ATP、Ca2＋-AT P和Mg2＋-ATP酶活性。结果显示,药物作用后大鼠大脑皮层和脑干Na＋、K＋-ATP酶活性与对照组比较降低显著（P〈0.05或P〈0.01）,小脑、脑干和海马Ca2＋-ATP酶活性与对照组比较显著降低（P〈0.05或P〈0.01）,大脑Mg2＋-AT P酶活性低于对照组（P〈0.05或P〈0.01）。结果表明,麻醉引起大鼠大脑皮层、脑干中Na＋-K＋-ATP酶活性降低,大脑皮层中Mg2＋-ATP酶活性低,小脑、脑干和海马脑区中Ca2＋-ATP酶活性降低可能是鹿特异性复合麻醉剂麻醉作用的机理之一。     

不同发育期大通牦牛组织线粒体COX、SDH活性的测定

为了探讨不同发育期牦牛组织线粒体标志酶氧化供能的作用,试验采用酶联免疫分析法(ELISA)对青海省大通种牛场1日龄、1月龄、3月龄、6月龄犊牦牛心肌、骨骼肌、肝脏、大脑组织线粒体细胞色素C氧化酶(COX)、琥珀酸脱氢酶(SDH)活性进行测定。结果表明:大通牦牛心肌、骨骼肌、肝脏、大脑组织线粒体COX、SDH活性在4个年龄组间差异不显著(P0.05),肝脏组织线粒体COX、SDH活性均高于其他组织。在同一海拔梯度,大通牦牛在发育期心肌、骨骼肌、肝脏、大脑组织线粒体氧化磷酸化作用及有氧代谢能力保持在相对稳定的水平。     

小型猪特异性麻醉颉颃剂对大鼠不同脑区Ca~(2+),Mg~(2+)-ATP酶活性的影响

为探讨Ca2+,Mg2+-ATP酶在小型猪特异性麻醉颉颃剂催醒过程中的作用,144只Wistar大鼠随机分为对照组和试验组,2组再分为早期催醒组、中期催醒组和晚期催醒组。用分光光度法测定不同脑区Ca2+,Mg2+-ATP酶活性。结果显示大鼠在不同时期注射小型猪特异性麻醉颉颃剂后,大鼠不同脑区的Ca2+,Mg2+-ATP酶活性均被激活,且Ca2+,Mg2+-ATP酶活性的这种变化趋势与大鼠行为学的变化相一致。结果表明小型猪特异性麻醉颉颃剂催醒作用可能与激活脑干和海马等脑区的Ca2+,Mg2+-ATP酶活性相关。     

荷斯坦牛血红细胞Na~+-K~+-ATP酶活力与其耐热性的相关性研究

[目的]为探讨Na+-K+-ATP酶在荷斯坦牛热应激过程中的作用.[方法]本研究比较测定了329头荷斯坦牛在非热应激期和热应激期的体温、红细胞K+和红细胞Na+-K+-ATP酶活力的变化.[结果]显示,试验牛群红细胞Na+-K+-ATP酶活力呈正态分布;不同胎次、产犊季节、泌乳期、场次牛Na+-K+-ATP酶活力差异不显著(P>0.05);不同性别牛Na+-K+-ATP酶活力差异极显著(P<0.01).在热应激期,同一泌乳期牛红细胞Na+-K+-ATP酶活力与产奶量下降率成负相关;荷斯坦牛的体温明显升高,产奶量明显下降,红细胞K+无显著变化,红细胞Na+-K+-ATP酶活力明显下降,与非热应激期相比只有红细胞K+差异不显著(P>0.05),其余都差异极显著(P<0.01);红细胞Na+-K+-ATP酶与体温成负相关,与产奶量成正相关,与红细胞K+的相关性不显著.[结论]在热应激期,荷斯坦牛红细胞Na+-K+-ATP酶活力与耐热性成正相关,红细胞Na+-K+-ATP酶活力高的荷斯坦牛,耐热性好.     

不同海拔地区牦牛心肌、骨骼肌线粒体氧自由基代谢的研究

对青海省玛多和刚察两县牦牛心肌、骨骼肌组织中肌红蛋白(Mb),心肌、骨骼肌中线粒体总抗氧化能力(T-AOC),超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-PX),乳酸脱氢酶(LDH)活性及丙二醛(MDA)含量进行了测定。结果:玛多牦牛心肌、骨骼肌中Mb含量均高于刚察牦牛,但差异不显著(P0.05);玛多牦牛心肌、骨骼肌线粒体中T-AOC,SOD,GSH-PX酶活性均显著高于刚察牦牛(P0.05),而LDH活性显著低于刚察牦牛(P0.05),玛多牦牛骨骼肌和心肌线粒体MDA含量低于刚察牦牛(P0.05)。结论:在高原低氧环境下,牦牛心肌、骨骼肌线粒体抗氧化能力随海拔高度的升高而增强,表明牦牛对高原低氧环境具有显著的适应性。     

一日龄大通牦牛和平原黄牛骨骼肌显微结构的比较

为了探讨1日龄大通牦牛骨骼肌组织学特点及对低氧的适应,以平原黄牛为对照,利用光镜和计算机图像分析系统测定骨骼肌肌纤维直径、表面积密度;通过透射电镜比较骨骼肌线粒体的面数密度、面积密度、体积密度、平均体积等结构参数。结果显示,1日龄大通牦牛骨骼肌肌纤维直径显著细于1日龄平原黄牛,表面积密度明显大于平原黄牛,差异极显著(P<0.01);1日龄大通牦牛骨骼肌细胞中线粒体平均体积小于1日龄平原黄牛,并具有极显著差异(P<0.01);而1日龄大通牦牛骨骼肌细胞中线粒体体积密度、面积密度、面数密度均大于1日龄平原黄牛,且差异极显著(P<0.01)。以上结果表明,大通牦牛通过增加骨骼肌线粒体面数密度、面积密度、体积密度,降低线粒体平均体积来提高其在低氧环境中对氧的利用,并且在长期进化中形成了肌纤维直径小,表面积密度大的组织学特点。     

铜缺乏对奶牛红细胞及组织细胞膜ATP酶活性的影响

为探讨铜缺乏对奶牛红细胞及组织细胞膜ATP活性的影响，在调查铜缺乏奶牛水土食物链微量元素状态的基础上，首次测度了病区6头发病犊奶牛（Ⅱ组）及该地区20头同年生临床健康奶牛（IH组，血清铜低于0.56mg/kg）红细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶及Mg^2 -ATP酶的活性，对20头IH组奶牛进行为期80d的补铜试验（含铜40mg/kg干饲料），以20头健区同年生健康奶犊牛作为对照（HH组，全血铜大于０.85mg/L）。同时测试了剖杀后６头病牛及健康牛组织细胞膜ＡＴＰ酶的活性。结果表明：Ⅱ组奶牛红细胞膜ＡＴＰ酶活性显著低于ＨＨ组及ＩＨ组。补铜后４０d，ＩＨ组奶牛红细胞膜Na^ -K^ -ATP酶均显著升高，病牛肝、脾、肾、大脑、小脑、淋巴结等组织细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶活性显著低于对照组。结论：铜缺乏较为严重地影响了奶牛红细胞及组织膜ＡＴＰ酶的活性，抑制了这些组织的正常生理功能。     

大通牦牛骨骼肌胶原纤维含量增龄性变化特点

为探讨高原牦牛随着年龄的增长,骨骼肌中胶原纤维含量的变化规律及其对高原低氧环境的适应性。本研究以不同生长期大通牦牛作为研究对象,并以平原黄牛作对照,对生长发育过程骨骼肌中胶原纤维含量变化进行测量。结果表明,大通牦牛骨骼肌中胶原纤维含量从1日龄的一个相对低点,到30日龄迅速增长至一个峰值,然后再逐渐下降,至成年时,胶原纤维含量降至最低。平原黄牛骨骼肌中胶原纤维含量1日龄处于峰值,与其他年龄组差异显著(P<0.05),而随着年龄的增长,胶原纤维含量下降,30日龄、180日龄和成年组差异不显著(P>0.05);相同发育阶段大通牦牛骨骼肌中胶原纤维含量均高于平原黄牛胶原纤维含量,且差异显著(P<0.05)。结果提示,大通牦牛骨骼肌中胶原纤维具有适应高原环境的组织学特点,但也在个体发育过程中,不断发生改变,以满足机体生长发育的需要和对外界环境的适应。     

Ca2+ ATPase in Dutch warmblood foals compared with Na+, K+ ATPase: intermuscular differences and the effect of exercise

We studied the effects of exercise without or with a subsequent period on pasture on Ca2+ ATPase concentration in foal skeletal muscle, and compared the results with those previously reported on Na+, K+ ATPase. Ca2+ ATPase was measured in homogenates as Ca2+-dependent steady-state phosphorylation from [gamma-32P]ATP. From day 7 after birth, 24 foals were divided into three groups: (i) staying in a box stall (Box); (ii) staying in a box stall with an exercise programme of an increasing number of sprints per day (Exercise); and (iii) staying on pasture (Pasture). Half of the foals (12 with four in each treatment group) were killed after 5 months. The remaining foals stayed on pasture until 11 months. In the 5-month Pasture group, Ca2+ ATPase concentration was 29.4 +/- 4.3 nmol/g wet weight (wt) (n = 4) in gluteus medius muscle, 25.2 +/- 3.3 nmol/g wet wt (n = 4) in semitendinosus muscle (both mixed fibre type), and 4.1 +/- 1.7 nmol/g wet wt (n = 3) in the slow masseter muscle. These values were not altered by exercise or by box rest. This was in contrast to the Na+, K+ ATPase concentration which was not different between the three muscles, but showed a 20% rise in gluteus medius and semitendinosus muscle after exercise. In the period from 5 to 11 months on pasture, there was no change in Ca2+ ATPase in any group. In conclusion, the Ca2+ ATPase concentration in foal muscle is around 6-fold higher in mixed fibres than in slow fibres. Furthermore, the enzyme is not up- or down-regulated by sprint exercise or subsequent rest.     

Calcium regulation by skeletal muscle membranes of horses with recurrent exertional rhabdomyolysis

OBJECTIVE: To determine whether an alteration in calcium regulation by skeletal muscle sarcoplasmic reticulum, similar to known defects that cause malignant hyperthermia (MH), could be identified in membrane vesicles isolated from the muscles of Thoroughbreds with recurrent exertional rhabdomyolysis (RER). SAMPLE POPULATION: Muscle biopsy specimens from 6 Thoroughbreds with RER and 6 healthy (control) horses. PROCEDURES: RER was diagnosed on the basis of a history of > 3 episodes of exertional rhabdomyolysis confirmed by increases in serum creatine kinase (CK) activity. Skeletal muscle membrane vesicles, prepared by differential centrifugation of muscle tissue homogenates obtained from the horses, were characterized for sarcoplasmic reticulum (SR) activities, including the Ca2+ release rate for the ryanodine receptor-Ca2+ release channel, [3H]ryanodine binding activities, and rate of SR Ca2+-ATPase activity and its activation by Ca2+. RESULTS: Time course of SR Ca2+-induced Ca2+ release and [3H]ryanodine binding to the ryanodine receptor after incubation with varying concentrations of ryanodine, caffeine, and ionized calcium did not differ between muscle membranes obtained from control and RER horses. Furthermore, the maximal rate of SR Ca2+-ATPase activity and its affinity for Ca2+ did not differ between muscle membranes from control horses and horses with RER. CONCLUSIONS AND CLINICAL RELEVANCE: Despite clinical and physiologic similarities between RER and MH, we concluded that RER in Thoroughbreds does not resemble the SR ryanodine receptor defect responsible for MH and may represent a novel defect in muscle excitation-contraction coupling, calcium regulation, or contractility.     

敌百虫对大鼠肝细胞[Ca~(2+)]_i、ATP酶及GJIC的影响

通过给大鼠肝细胞培养液中加入敌百虫(染毒终质量浓度分别为0,2,10,50 mg/L),探讨敌百虫对肝细胞内钙离子([Ca2+]i)、Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性及间隙连接通讯(GJIC)的影响。结果显示,不同质量浓度敌百虫作用12 h,[Ca2+]i均极显著高于对照组(P0.01);随着染毒剂量的增加,Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性及GJIC均显著或极显著下降(P0.05或P0.01)。说明敌百虫对肝细胞有明显的毒性作用,GJIC的下调可能与Ca2+浓度的升高和Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性的下降有关。     

秋季断犊母牦牛促发情试验

在青海省大通种牛场育种一队对27头秋季断犊母牦牛进行了促发情试验。母牦牛断犊并经21-23d恢复饲养后,采用两次PG注射法和GnRH+PG注射法对母牦牛进行同期发情处理,结果显示:试验I组母牦牛发情6头,发情率66.67%,试验II组母牦牛发情2头,发情率22.22%。经统计分析,试验I组与试验II组母牦牛之间发情率差异不显著(P﹥0.05),但显著高于对照组(P﹤0.05),试验II组母牦牛发情率与对照组间差异不显著(P﹥0.05)。     

Effects of training on potassium homeostasis during exercise and skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged Dutch Warmblood horses

OBJECTIVE: To investigate the effects of moderate short-term training on K+ regulation in plasma and erythrocytes during exercise and on skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged horses. ANIMALS: Four 4- to 6-year-old and four 10- to 16-year-old Dutch Warmblood horses. PROCEDURE: The horses underwent a 6-minute exercise trial before and after 12 days of training. Skeletal muscle Na+,K(+)-ATPase concentration was analyzed in gluteus medius and semitendinosus muscle specimens before and after the 12-day training period. Blood samples were collected before and immediately after the trials and at 3, 5, 7, and 10 minutes after cessation of exercise for assessment of several hematologic variables and analysis of plasma and whole-blood K+ concentrations. RESULTS: After training, Na+,K(+)-ATPase concentration in the gluteus medius, but not semitendinosus, muscle of middle-aged horses increased (32%), compared with pretraining values; this did not affect the degree of hyperkalemia that developed during exercise. The development of hyperkalemia during exercise in young adult horses was blunted (albeit not significantly) without any change in the concentration of Na+,K(+)-ATPase in either of the muscles. After training, the erythrocyte K+ concentration increased (7% to 10%) significantly in both groups of horses but did not change during the exercise trials. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, the activation of skeletal muscle Na+,K(+)-ATPase during exercise is likely to decrease with age. Training appears to result in an increase in Na+,K(+)-ATPase activity in skeletal muscle with subsequent upregulation of Na+,K(+)-ATPase concentration if the existing Na+,K(+)-ATPase capacity cannot meet requirements.     

Specificity and reversibility of the training effects on the concentration of Na+,K+-Atpase in foal skeletal muscle

The purpose of the present study was to determine whether training and detraining affect the Na+,K+-ATPase concentration in horse skeletal muscles, and whether these effects are specific for the muscles involved in the training programme. Twenty-four Dutch Warmblood foals age 7 days were assigned randomly to 3 groups: Box (box-rest without training), Training (box-rest with training: short-sprint) and Pasture (pasture without training). Exercise regimens were carried out for 5 months and were followed by 6 months of detraining. Five of the foals in each group were subjected to euthanasia at age 5 months and the remaining foals at 11 months. Muscle samples were collected from the deep part of the gluteus medius, semitendinosus and masseter muscles. The Na+,K+-ATPase concentration was quantified by [3H]ouabain binding. In the Training group, the concentration of Na+,K+-ATPase in gluteus medius and semitendinosus muscle, but not in masseter muscle, showed a relative increase of 20% (P<0.05) as compared to Box foals. After detraining for the subsequent 6 months, the concentration of Na+,K+-ATPase in semitendinosus muscle remained the same, while that in gluteus medius muscle was reduced by 10%. It is concluded that: 1) short-sprint training for 5 months induced an increase of the Na+,K+-ATPase concentration in gluteus medius and semitendinosus muscles of the foal. Interestingly, this effect persisted during the 6 months of the detraining period. Whether the higher Na+,K+-ATPase concentration due to training of young foals leads to a better athletic performance when they become mature still needs to be established; 2) the factors that initiate an increase in Na+,K+-ATPase concentration following training are likely to be located in the muscle itself and 3) the training effect may last for several months after returning to normal activity, especially in muscles containing a high percentage of fast-twitch fibres.