根田鼠冷驯化过程中的适应性产热特征

对栖居青海高原的根田鼠于冷驯化１，７和２１天的非颤抖性产热，褐色脂肪组织和 脏的组织蛋白和线粒体蛋白含量，心脏和肌肉的线粒体蛋白含量，四种组织线粒 的细胞色素氧化酶活力及肝脏线粒体的状态Ⅳ呼吸能力等指标进行了测定。     

冷暴露对中缅树鼩肝脏、膈肌和心肌线粒体呼吸的影响

在4℃急性冷暴露(1 h,4 h,8 h,24 h)和持续冷暴露 (7 d,14 d,28 d)条件下,测定中缅树鼩膈肌、心肌和肝脏的线粒体状态Ⅲ呼吸、状态Ⅳ呼吸、呼吸控制率(RCR)、线粒体蛋白含量以及肝脏线粒体P/O值的变化.结果表明:肝脏线粒体状态Ⅲ、状态Ⅳ呼吸随着低温处理时间的延长,呼吸速率均极显著增加,在28 d后分别增加了132.9%、124.4%(P<0.01),RCR与对照比较,在8 h和7 d组分别显著增加了35.8%和48.4%(P<0.05),线粒体蛋白含量也极显著增加,在28 d后增加了104.7%(P<0.01),P/O值在整个低温处理过程中呈下降趋势,在28 d后降低了40.2%,达到极显著水平(P<0.01);膈肌线粒体状态Ⅲ呼吸在整个低温处理期间没有显著变化,状态Ⅳ呼吸在28 d达到极显著增加(P<0.01,64.9%),RCR在28 d后显著降低(P<0.05, 42.1%),线粒体蛋白只有4 h组有极显著增加(P<0.01,45.2%);心肌的状态Ⅲ呼吸在8 h组有着极显著的增加(P<0.01, 54.7%),状态Ⅳ呼吸随着低温处理时间的增加而显著增加,28 d后增加了94.7%(P<0.01),RCR在28 d后降低37.8%(P<0.01),线粒体蛋白表现出先下降再上升的趋势,8 h组下降37.8%(P<0.01),28 d增加25.2%(P<0.05).说明中缅树鼩在冷胁迫的条件下肝脏线粒体呼吸能力显著增强,主要表现为状态Ⅳ呼吸即质子漏产热的显著增强,膈肌和心肌的线粒体呼吸也具有一定的适应性变化,补偿了冷胁迫条件下中缅树鼩增加的能量需求,是中缅树鼩在冷胁迫中重要的适应对策.     

布氏田鼠对低温的适应性产热

将布氏田鼠置１到３０天的低温（６±１℃）环境下暴露,其体重与体温没有明显变化;静止代谢率（ＲＭＲ）、非震颤性产热（ＮＳＴ）及血清中三碘甲腺原氨酸（Ｔ３）含量,随冷暴露时间的延长而增加;肝线粒体状态３呼吸及细胞色素Ｃ氧化酶活力亦随冷暴露时间的延长而升高。肝线粒体蛋白质含量经冷暴露３０天时有明显升高,表明动物在低温适应中肝线粒体蛋白合成增加与呼吸功能增强是个体ＲＭＲ提高的细胞学机制之一。布氏田鼠褐色脂     

光周期和温度对中缅树()产热能力的影响

低温短光照诱导中缅树()体重增加、体温降低、产热能力增强,静止代谢率增加的速度和比例高于非颤抖性产热;低温短光照刺激肝脏线粒体蛋白质量、线粒体氧化能力和细胞色素C氧化酶活性显著增加;短光照也刺激褐色脂肪组织总蛋白和线粒体蛋白量、线粒体状态IV呼吸、细胞色素C氧化酶、α-磷酸甘油氧化酶及T45'-脱碘酶活性显著提高.低温和短光照促使血清T3浓度增加、T4浓度不变.结果表明,中缅树()的适应性产热特征不仅受到低温的显著影响,而且光周期也参与了适应性产热的调节.     

非冬眠期达乌尔黄鼠对低温的适应性产热

达乌尔黄鼠在夏季冷驯化低温环境下（５±１℃）暴露４周后,导致脂肪积累的抑制;其静止代谢率（ＲＭＲ）和非颤抖性产热（ＮＳＴ）分别提高了１２％和８５％;肝脏线粒体的蛋白含量、状态３呼吸和细胞色素Ｃ氧化酶活力分别提高了２２％、４２％和４８％,表明在低温环境下肝脏线粒体产热能力的增强是ＲＭＲ提高的细胞学机制之一;褐色脂肪组织（ＢＡＴ）的重量、蛋白含量、细胞色素Ｃ氧化酶活力和α－磷酸甘油氧化酶活力极显著提高,表明在低温环境下ＢＡＴ是达乌尔黄鼠的主要产热器官,ＢＡＴ产热能力的增强是ＮＳＴ提高的重要细胞学机制之一。冷驯化没有引起达乌尔黄鼠血清Ｔ３、Ｔ４含量的变化     

冷驯化对大绒鼠和高山姬鼠肝脏线粒体呼吸的影响

大绒鼠Eothenomys miletus和高山姬鼠Apodemus chevrieri为横断山地区小型哺乳动物的代表.分别测定大绒鼠和高山姬鼠冷驯化组(7 d、14 d、21 d、28 d)与对照组(0 d)中肝脏线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ、呼吸磷氧比的变化以及较大绒鼠和高山姬鼠的相应项,探讨大绒鼠和高山姬鼠对不同冷环境的适应情况以及肝脏产热的机理.结果 表明:在冷驯化条件下,大绒鼠和高山姬鼠的产热显著增加,无论是大绒鼠还是高山姬鼠实验组比对照组肝脏线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ、呼吸磷氧比有着显著的提高,在两者的比较中,在同等冷驯化条件下高山姬鼠增加的程度高于大绒鼠.     

高原鼠兔冷驯化和脱冷驯化中的产热变化

本文探讨了冷驯化和脱冷驯化对高原鼠兔代谢产热的影响。在冷驯化中，鼠兔的静止代谢率逐渐增加，血清Ｔ３‘／Ｔ４比率上升，肝线粒体状态３呼吸明显增加，线粒体细胞色素Ｃ氧化酶激活，而肝脏线粒体蛋白含量没有明显变化。     

冷暴露对中缅树鼩褐色脂肪组织中解偶联蛋白1含量的影响

自备抗血清采用酶联免疫法测定了中缅树鼩(Tupaia belangeri)在(5±1)℃冷暴露0 d、7 d、14 d、21d、28 d时,褐色脂肪组织(BAT)中解偶联蛋白1(UCP1)的含量.结果表明,随着冷暴露时间的延长,中缅树鼩的体重、褐色脂肪组织重量均表现出了增加的趋势,BAT线粒体总蛋白和UCP1的含量也呈增加的趋势,其中UCP1的含量在28 d时达到极显著水平,比对照组增加了55.9%.说明冷暴露能够诱导中缅树鼩UCP1表达增加,从而使其适应性产热增加.     

光周期和温度对中缅树Qu产热能力的影响

低温短光照诱导中缅树Ｑｕ体重增加、体温降低、产热能力增强,静止代谢率增另的速度和比例高于非颤抖性产热,低温短光照刺激肝脏一体蛋白质量、线粒体氧化能力和细胞色素Ｃ氧化酶活性显著增加;短光照也刺激褐色脂肪组织总蛋白和线粒体蛋白量、线粒体状态ＩＶ呼吸、细胞色素Ｃ氧化酶、α－磷酸甘油氧化酶及Ｔ４５－脱碘酶活性显著提高。低温和短光照促使血清Ｔ３浓度增加、Ｔ４浓度不变。结果表明,中缅树Ｑｕ的适应性产热特征不仅     

中缅树鼩的非颤抖性产热及细胞呼吸特征

中缅树鼩(Tupaia belangeri chinensis)是东南亚树鼩中分布最北的一个种。在热中性区内的非颤抖性产热(nonshivering thermogenesis NST)分别为2.57±0.21(冬)和2.21±0.12(夏)mlO_2/(g·h);分别为体重预期值的75.9％和61.2％,两者不仅冬季显著高于夏季,而且亦高于典型的热带种类,但低于温带类群。褐色脂肪组织(brown adipose tissue,BAT)的重量冬季为0.622±0.015 S,夏季0.532±0.80 g,冬季也显著高于夏季;同时,BAT总蛋白含量、线粒体蛋白含量以及细胞α-磷酸甘油氧化酶和细胞色素C氧化酶活性,冬季也显著高于夏季,但增加的比例较温带种类低;而肝脏细胞的上述指标及线粒体状态Ⅲ、状态Ⅳ呼吸等,冬夏两季均无显著差异。因此,中缅树鼩的NST和细胞产热能力介于热带与温带类群之间,显示出向温带类型过渡的趋势。     

The role of insulin in norepinephrine turnover and thermogenesis in brown adipose tissue after acute cold-exposure

The role of insulin in norepinephrine turnover (NE) and thermogenesis in brown adipose tissue (BAT) after acute cold-exposure was studied using streptozocin (STZ)-induced diabetic rats. NE turnover was estimated by the NE synthesis inhibition technique with alpha-methyl-p-tyrosine. BAT thermogenesis was estimated by measuring mitochondrial guanosine-5'-diphosphate (GDP), cytochrome oxidase activity and mitochondrial oxygen consumption in BAT at an ambient temperature of 22 degrees C and during a six-hour cold-exposure at 4 degrees C. In insulin-deficient diabetic rats, the NE turnover, mitochondrial GDP binding, cytochrome oxidase activity and mitochondrial oxygen consumption in BAT at 22 degrees C were significantly reduced, compared with those of control rats. Treatment of STZ-induced diabetic rats with insulin prevented a decrease in NE turnover and BAT thermogenesis. Acute cold-exposure increased the NE turnover of BAT in insulin-deficient diabetic rats. The BAT thermogenic response to acute cold-exposure, however, did not occur in insulin-deficient diabetic rats. These results suggest that insulin is not essential in potentiating NE turnover in BAT after acute cold-exposure, but is required for cold-induced thermogenesis.     

Adaptive thermogenesis in Brandt's vole (Lasiopodomys brandti) during cold and warm acclimation

Brandt's voles (Lasiopodomys brandti) exposed to cold (5±1 °C) or warm (23±1 °C) showed some physiological and biochemical variations which might be important in adaptation to their environments. Cold acclimation induced increases in resting metabolic rate (RMR) and the serum triiodothyronine (T₃) level, the state-4 respiration of liver and muscle mitochondria were activated after 7 days when animals exposed to cold, and the activity of cytochrome c oxidase (COX) of liver and muscle mitochondria tended to rise with cold exposure. RMR and T₃ level decreased during warm acclimation. The state-4 respiration of liver mitochondria declined after 3 days and muscle after 7 days when animals exposed to warm, and the activities of COX of liver and muscle mitochondria tended to decrease with warm acclimation. The cold activation of liver and muscle mitochondrial respiration (regulated by T₃) was one of the cytological mechanisms of elevating RMR. Both state-4 respiration and COX activity of brown adipose tissue (BAT) mitochondria increased significantly during cold acclimation and decreased markedly after acclimated to warm. The uncoupling protein 1 (UCP1) contents in BAT increased after exposure to cold and decreased after warm acclimation. Nonshivering thermogenesis (NST) plays an important role in the process of thermoregulation under cold acclimation for Brandt's voles. Changes in thermogenesis is a important way to cold adaptation for Brandt's voles in natural environments.     

The consequences of acute cold exposure on protein oxidation and proteasome activity in short-tailed field voles,microtus agrestis

During cold exposure, animals upregulate their metabolism and food intake, potentially exposing them to elevated reactive oxygen species (ROS) production and oxidative damage. We investigated whether acute cold (7 +/- 3 degrees C) exposure (1, 10, or 100 h duration) affected protein oxidation and proteasome activity, when compared to warm controls (22 +/- 3 degrees C), in a small mammal model, the short-tailed field vole Microtus agrestis. Protein carbonyls and the chymotrypsin-like proteasome activity were measured in plasma, heart, liver, kidney, small intestine (duodenum), skeletal muscle (gastrocnemius), and brown adipose tissue (BAT). Trypsin-like and peptidyl-glutamyl-like proteasome activities were determined in BAT, liver, and skeletal muscle. Resting metabolic rate increased significantly with duration of cold exposure. In skeletal muscle (SM) and liver, protein carbonyl levels also increased with duration of cold exposure, but this pattern was not repeated in BAT where protein carbonyls were not significantly elevated. Chymotrpsin-like proteasome activity did not differ significantly in any tissue. However, trypsin-like activity in SM and peptidyl-glutamyl-like activity in both skeletal muscle and liver, were reduced during the early phase of cold exposure (1-10 h), correlated with the increased carbonyl levels in these tissues. In contrast there was no reduction in proteasome activity in BAT during the early phase of cold exposure and peptidyl-glutamyl-like activity was significantly increased, correlated with the lack of accumulation of protein carbonyls in this tissue. The upregulation of proteasome activity in BAT may protect this tissue from accumulated oxidative damage to proteins. This protection may be a very important factor in sustaining uncoupled respiration, which underpins nonshivering thermogenesis at cold temperatures.     

达乌尔黄鼠产热的季节性变化

达乌尔黄鼠（Ｃｉｔｅｌｌｕｓｄａｕｒｉｃｕｓ）的产热表现出明显的季节性变化。在非冬眠期,静止代谢率（ＲＭＲ）和非颤抖性产热（ＮＳＴ）于春季最高,秋季次之,夏季最低。冬眠期,ＲＭＲ降到极低水平,只为春季的３．０％。肝脏的线粒体蛋白含量、线粒体呼吸和细胞色素Ｃ氧化酶活力在秋季显著高于其它各季。褐色脂肪组织（ＢＡＴ）的重量、线粒体蛋白含量、细胞色素Ｃ氧化酶活力和α－磷酸甘油氧化酶活力,在夏季处于一年中的最低水平,到了冬季这些指标达到一年中的最高水平。在非冬眠季节ＢＡＴ产热能力升高时,ＮＳＴ能力也相应升高,这表明ＢＡＴ产热能力的增强是ＮＳＴ能力提高的部分机制。达乌尔黄鼠血清Ｔ＿４含量在年周期中没有明显改变,冬眠时血清Ｔ＿３含量显著高于其它各季。     

长爪沙鼠褐色脂肪组织和肝脏产热特征的季节性变化

长爪沙鼠（Meriones unguiculatus）是一种栖息于典型草原和荒漠草原非冬眠群居性的小型哺乳动物。为研究其产热功能的季节变化,我们分别在2003年秋季（9月下旬）、冬季（11月下旬）、2004年春季（3月底-4月下旬）和夏季（7月下旬）,分别测定了其体重、褐色脂肪组织和肝脏的重量、线粒体总蛋白含量和细胞色素c氧化酶活力,以及褐色脂肪组织中解偶联蛋白1（Uncoupling protein1,UCPl）的含量等。结果显示：除雄鼠的体重显著高于雌鼠外,其它各项指标均无性别差异。体重和褐色脂肪组织的重量都在冬季较高,显著高于夏季,而肝脏的重量在夏季显著高于其它季节。褐色脂肪组织和肝脏的线粒体蛋白含量和细胞色素c氧化酶活力以及UCP1含量,都在冬季较高,夏季较低。这些结果表明：在野外条件下,褐色脂肪组织和肝脏在细胞水平上产热能力的提高和UCP1含量的增加,是长爪沙鼠抵御寒冷的重要方式。     

Vitamin E attenuates cold-induced rat liver oxidative damage reducing H2O2 mitochondrial release

Vitamin E is a major chain-breaking antioxidant which is able to reduce liver oxidative damage without modifying aerobic capacity in T(3)-treated rats. We investigated whether vitamin E has similar effects in hyperthyroid state induced by cold exposure. Cold exposure increased aerobic capacity and O(2) consumption in homogenates and mitochondria and tissue mitochondrial protein content. Vitamin E did not modify aerobic capacity and mitochondrial protein content of cold liver, but increased ADP-stimulated respiration of liver preparations. Succinate-supported H(2)O(2) release rates were increased by cold during basal and stimulated respiration, whereas the pyruvate/malate-supported ones increased only during basal respiration. Vitamin administration to cold-exposed rats decreased H(2)O(2) release rates with both substrates during basal respiration. This effect reduced ROS flow from mitochondria to cytosol, limiting liver oxidative damage. Cold exposure also increased mitochondrial capacity to remove H(2)O(2), which was reduced by vitamin treatment, showing that the antioxidant also lowers H(2)O(2) production rate. The different effects of cold exposure and vitamin treatment on H(2)O(2) generation were also found in the presence of respiration inhibitors. Although this can suggest that the cold and vitamin induce opposite changes in mitochondrial content of autoxidizable electron carriers, it is likely that vitamin effect is due to its capacity to scavenge superoxide radical. Finally, vitamin E reduced mitochondrial oxidative damage and susceptibility to oxidants, and prevented Ca(2+)-induced swelling elicited by cold. In the whole, our results suggest that vitamin E is able to maintain aerobic capacity and attenuate oxidative stress of hepatic tissue in cold-exposed rats modifying mitochondrial population characteristics.     

The molecular and biochemical basis of nonshivering thermogenesis in an African endemic mammal, Elephantulus myurus

Uncoupling protein 1 (UCP1) mediated nonshivering thermogenesis (NST) in brown adipose tissue (BAT) is an important avenue of thermoregulatory heat production in many mammalian species. Until recently, UCP1 was thought to occur exclusively in eutherians. In the light of the recent finding that UCP1 is already present in fish, it is of interest to investigate when UCP1 gained a thermogenic function in the vertebrate lineage. We elucidated the basis of NST in the rock elephant shrew, Elephantulus myurus (Afrotheria: Macroscelidea). We sequenced Ucp1 and detected Ucp1 mRNA and protein restricted to brown fat deposits. We found that cytochrome c oxidase activity was highest in these deposits when compared with liver and skeletal muscle. Consistent with a thermogenic function of UCP1 isolated BAT mitochondria showed increased state 4 respiration in the cold, as well as palmitate-induced, GDP-sensitive proton conductance, which was absent in liver mitochondria. On the whole animal level, evidence of thermogenic function was further corroborated by an increased metabolic response to norepinephrine (NE) injection. Cold acclimation (18 degrees C) led to an increased basal metabolic rate relative to warm acclimation (28 degrees C) in E. myurus, but there was no evidence of additional recruitment of NE-induced NST capacity in response to cold acclimation. In summary, we showed that BAT and functional UCP1 are already present in a member of the Afrotheria, but the seasonal regulation and adaptive value of NST in Afrotherians remain to be elucidated.     

布氏田鼠胎后发育过程中褐色脂肪组织和肝脏的产热特征

为探讨布氏田鼠胎后恒温能力的发育过程,本文测定了1、5、9、17、21、33 和41 日龄幼体的褐色脂肪组织(BAT)和肝脏的重量、线粒体蛋白含量和细胞色素c氧化酶(COX) 的活性。布氏田鼠胎后发育期间BAT 增补明显,主要表现为重量的增加和单位组织重量COX 活性的升高等,属典型的晚成型发育特征。布氏田鼠胎后发育过程中BAT 和肝脏产热特征的变化与幼体的产热特点和恒温能力的发育是相一致的。     

Leptin-programmed rats respond to cold exposure changing hypothalamic leptin receptor and thyroid function differently from cold-exposed controls

We showed that neonatal leptin treatment programmes for hyperleptinemia and central leptin resistance both at 30days-old and adulthood, while programmes for lower serum T3 at 30days-old, but higher thyroid hormones (TH) at adulthood. As in these animals, acute cold at 30days-old normalized leptinemia and restored the expression of hypothalamic leptin receptor (OBR), here we evaluate the effect of cold exposure on the thyroid function and OBR in adult rats programmed by neonatal hyperleptinemia. Pups were divided into 2 groups: Lep-injected with leptin (8μg/100g/BW, sc) for the first 10days of lactation, and C-injected with saline. At 150days, both groups were subdivided into: LepC and CC, which were exposed to 8°C for 12h. Serum leptin, TH, TSH, liver type I and brown adipose tissue (BAT) type II deiodinases (D1 and D2) activities, liver mitochondrial alpha-glycerol-3-phosphate dehydrogenase (mGPD) activity and adrenal catecholamine content were measured. Hypothalamic and thyroid OBR protein contents were evaluated. Differences were significant when p<0.05. Lep group had hyperleptinemia (+19%), higher T4 (+20%) and T3 (+30%) with lower TSH (-55%), higher liver D1 (1.4 fold-increase), lower BAT D2 (-44%) and liver mGPD activities (-55%), higher adrenal catecholamines (+44%), lower hypothalamic OBR (-51%) and normal thyroid OBR. Cold exposure normalized leptinemia, D1, mGPD, catecholamine and hypothalamic OBR. However, cold exposure further increased TH and decreased D2. Thus, cold exposure normalizes most of the changes programmed by neonatal hyperleptinemia, at the expense of worsening the hyperthyroidism and BAT thermogenesis.     

布氏田鼠冷暴露中褐色脂肪组织的增补及解偶联蛋白基因表达

布氏田鼠 (Microtusbrandti)随机分组暴露在冷环境 [12L∶12D ,(4± 2 )℃ ]中 12h ,1d ,3d ,7d ,14d ,2 1d和 2 8d ;对照组生活在温暖环境下 [12L∶12D ,(2 5± 2 )℃ ]。与对照组相比 ,布氏田鼠的褐色脂肪组织 (BAT)重量在冷暴露 12h～ 3d时降低 ,7～ 2 1d时则增加。 7～ 2 8d冷暴露组动物的BAT总蛋白和总DNA含量均比对照组明显提高。冷环境中的布氏田鼠解偶联蛋白 (UCP)的mRNA随时间的延长而表达上调 ,在冷暴露 2 1d时达到高峰。结果表明 ,冷暴露能够诱导布氏田鼠BAT细胞增补和UCP基因表达 ,从而使适应性产热增加。