冷驯化对中缅树鼩PPARα、COXⅡ及PGC-1α基因表达量的影响

为了研究冷驯化条件下中缅树鼩Tupaia belangeri的脂肪组织是否会转化,本研究测定了冷驯化条件下中缅树鼩脂肪组织质量,脂肪转化因子过氧化物酶体增殖激活受体α(PPARα)、环氧化酶-2(COXⅡ)及过氧化物酶体增殖物受体γ共激活因子1α(PGC-1α)基因表达量的变化。结果表明:中缅树鼩冷驯化组无论是褐色脂肪组织(BAT)质量,还是大网膜白色脂肪组织(WAT)质量均较对照组显著增加(P0.01),脂肪转化因子PPARα、COXⅡ及PGC-1α基因表达量也显著上调(P0.01)。以上结果说明中缅树鼩WAT中PPARα、COXⅡ、PGC-1α基因表达量的增加可能诱导了WAT细胞褐变,进而向BAT细胞转化和提高BAT中解偶联蛋白1的表达。     

冷驯化及复温对中缅树鼩白色脂肪组织中Ucp1和Cd137表达的影响

为探讨中缅树鼩(Tupaia belangeri) 白色脂肪组织（white adipose tissue, WAT）应对环境温度变化的适应机制,以荧光标记的Ucp1和Cd137抗体分别对中缅树鼩WAT进行标记,利用流式细胞术分选分析出对照组、冷驯化28d组、复温28d组阳性的白色脂肪细胞。结果发现：WAT中表达Ucp1、Cd137阳性的白色脂肪细胞百分数,在冷驯化组均显著高于对照组,而复温组均显著低于冷驯化组,恢复至对照组水平。表明：冷驯化诱导中缅树鼩WAT中表达Ucp1、Cd137的阳性细胞群增加,显示冷驯化能诱导中缅树鼩的WAT发生褐变;复温后表达Ucp1、Cd137阳性的白色脂肪细胞百分数恢复到对照水平,反映出WAT的可塑性。因此,WAT的可塑性调节也是中缅树鼩适应环境温度变化的重要产热机制。     

PRDM16和BMP7基因表达量在中缅树鼩体重调节中的作用

为探讨季节性环境变化下中缅树鼩PRDM16(PR domain-containing 16)和BMP7(bone morphogenetic proteins 7)基因表达量对其生理适应性调节的作用,本研究测定了野外不同季节和实验室冷驯化条件下中缅树鼩的体重、静止代谢率(resting metabolic rate,RMR)、非颤抖性产热(nonshivering thermogenesis,NST)、摄食量、PRDM16和BMP7基因表达量的变化。结果表明:季节性变化过程中中缅树鼩的体重、RMR、NST、褐色脂肪组织(brown adipose tissue,BAT)重、白色脂肪组织(white adipose tissue,WAT)重和摄食量均是冬季显著高于夏季;中缅树鼩不同季节的PRDM16和BMP7基因表达量差异极显著,PRDM16表达量的季节变化趋势为:冬季秋季春季夏季;BMP7表达量的季节变化趋势为:冬季秋季夏季春季。冷驯化条件下,中缅树鼩的体重、RMR、NST、摄食量、BAT含量、大网膜WAT含量显著增加,PRDM16和BMP7基因表达量也显著增加。以上结果表明,中缅树鼩褐色脂肪细胞存在PRDM16和BMP7肌源性起源,即冬季或者低温条件下中缅树鼩PRDM16和BMP7表达量上调,促进褐色脂肪细胞形成,增加NST来弥补产热的不足,以适应冬季寒冷的环境。PRDM16和BMP7在中缅树鼩季节性产热调节和能量代谢中起着重要的作用。     

冷驯化对中缅树鼩褐色脂肪组织适应性产热的影响

目的探讨环境温度对中缅树鼩体重、褐色脂肪组织(BAT)产热活性及解偶联蛋白1含量的影响,为建立树鼩肥胖模型提供理论依据。方法将40只体重相似的成年中缅树鼩随机分为5组(每组8只):对照组(0 d),置于(25±1)℃,12 L/12 D条件下饲养;以及置于(5±1)℃,12 L/12 D条件下分别驯化7、14、21 d和28d组。驯化结束后测定动物的体重、非颤抖性产热(NST)、褐色脂肪组织重量以及解偶联联蛋白1(UCP1)的含量。结果与对照组(0 d)相比,冷驯化组中缅树鼩的体重、NST、BAT重量以及UCP1含量都显著增加,BAT颜色也明显加深,冷驯化28 d后,体重增加了26.32%,NST增加了20.65%,BAT重量增加了53.85%,UCP1含量增加了43%。UCP1含量与BAT重量和NST显著正相关。结论中缅树鼩可能通过冷驯化诱导BAT组织增生和UCP1表达上调,从而增强BAT产热活力以增加能量支出,推测BAT可能作为以能量学途径治疗肥胖的靶器官。     

高脂高糖食物诱导中缅树鼩肥胖的研究

为了研究高糖高脂食物诱导中缅树鼩肥胖的模型,在高脂高糖食物诱导条件下进行了中缅树鼩的血液指标测定、脂肪组织流式细胞分析和正电子发射计算机断层显像(positron emission tomography/computed tomography,PET/CT)扫描成像研究。结果表明:在28 d时中缅树鼩白色脂肪组织(white adipose tissue,WAT)质量是对照组的3.7倍,49 d时WAT质量是对照组的6.4倍;49 d时实验组树鼩的甘油三酯(triglyceride,TG)和高密度脂蛋白(high density lipoprotein,HDL)含量与对照组树鼩差异显著,血糖(blood glucose)水平、总胆固醇(total cholesterol,CHOL)和血小板(blood platelet,PLT)含量与对照组树鼩差异极显著;此外,食物诱导28 d组中缅树鼩腹部WAT和褐色脂肪组织(brown adipose tissue,BAT)的18氟-氟化脱氧葡萄糖(18-fluoro-fluorodeoxyglucose,~(18)F-FDG)吸收60 min后,平均辐射值(standardized uptake value,SUV)显著高于对照组,49 d时有所下降。流式细胞分析结果表明,28 d时WAT和BAT中R2群标记阳性显著增加,而49 d后有所降低。以上结果表明:食物诱导可以使中缅树鼩产生肥胖并显著增加中缅树鼩WAT的质量,中缅树鼩在28 d时会出现肥胖抵抗的现象,但是随着时间的延长,该抵抗现象逐渐减少。     

大理和勐腊中缅树鼩血清及 肝差异代谢物的比较

为探究不同海拔地区中缅树鼩(Tupaia belangeri)血清和肝的差异代谢物及代谢通路变化,本研究采集大理(高海拔)和勐腊(低海拔)中缅树鼩分别12只和8只的血清和肝,采用非靶向代谢组气相色谱-质谱联用检测技术测定代谢物。结果表明,高海拔种群和低海拔种群相比,血清中一共有36种代谢物差异显著,其中,柠檬酸、葡萄糖、胆固醇等共32种代谢物的浓度上调,N-乙酰谷氨酸、葵酸、庚酸和对羟基苯甲酸这4种代谢物的浓度下调;高海拔种群肝差异代谢物和低海拔肝相比,一共有18种代谢物浓度差异显著,其中,苹果酸、核糖、葡萄糖等共10种差异代谢物浓度上调,谷氨酰胺、乙醇酸、硬脂酸等共8种差异代谢物浓度下调。高海拔种群与低海拔种群相比,血清中一共有76条代谢通路活性得分差异显著,其中,69条活性得分上调,7条活性得分下调;肝中一共有75条代谢通路活性得分差异显著,其中,43条活性得分上调,32条活性得分下调。以上结果说明,中缅树鼩在面对不同环境时,会调节不同组织中不同代谢通路(如三羧酸循环、糖酵解、脂类代谢和氨基酸代谢)的代谢物含量来适应环境,且血清比肝对环境变化更敏感。     

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.     

中缅树鼩肝在冷适应条件下的产热特征

中缅树鼩(Tupaia belangeri)为东洋界特有小型哺乳动物。本研究测定了中缅树鼩冷驯化实验组(7d、14d、21d、28d)与对照组(0d)中肝线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ的变化,探讨中缅树鼩对不同冷环境的适应情况以及肝产热的机理。结果表明,在冷驯化条件下,中缅树鼩产热显著增加,与对照组相比,实验组肝总蛋白含量、肝线粒体蛋白含量、呼吸状态Ⅲ和状态Ⅳ有着显著的提高,在28d后分别增加了39.9%、39.3%、84.9%、181.1%,因此,肝在中缅树鼩冷适应产热过程中具有重要的作用。同时,从生理生态方面为树鼩的岛屿起源学说提供依据。     

冷暴露对中缅树鼩适应性产热特征的影响

在冷暴露(5 ± 1℃)1 d,7 d,14 d 和21d 对中缅树鼩的肝脏、心脏、膈肌、腓肠肌和褐色脂肪组织(BAT)的总蛋白和线粒体蛋白含量、线粒体的状态Ⅳ呼吸能力、细胞色素C 氧化酶活力及血清中甲状腺激素的水平等指标进行了测定。结果表明:冷暴露过程中肝脏线粒体蛋白含量对低温的反应比总蛋白的反应剧烈,心脏和BAT 组织的线粒体蛋白含量也随着冷驯化时间的延长而显著增加,但腓肠肌的反应较为温和;各组织线粒体的状态Ⅳ呼吸能力均显著增强,除腓肠肌外其它各组织细胞色素C 氧化酶活性也随冷暴露时间的延长而急剧增加。说明在低温条件下,肝脏、腓肠肌、心脏和BAT 等组织都参与了体温调节的过程,在自然生境中,低温是重要的刺激产热的调节因子。     

寒冷诱导产热时大鼠棕色脂肪能量代谢相关蛋白谱的变化

棕色脂肪组织(BAT)的生理作用与白色脂肪显著不同,它以产热的形式释放能量而不是将能量以ATP的形式储存．线粒体是在能量代谢和维持细胞稳态中具有重要功能的细胞器．为了更好地了解棕色脂肪中的能量代谢过程,运用双向电泳及质谱相结合的技术,分离了大鼠白色和棕色脂肪线粒体,对其差异蛋白质谱进行了系统分析和鉴定．参与脂肪和氨基酸代谢、三羧酸循环及线粒体呼吸链的蛋白质在棕色脂肪线粒体中的表达明显高于白色脂肪线粒体,在寒冷诱导下这些蛋白质的表达进一步上调．此外,参与辅酶Q合成的一系列COQ 基因在棕色脂肪中经寒冷适应后表达明显上调．该研究表明,辅 酶Q合成的增高在非颤栗性产热中具有重要作用,为进一步了解棕色脂肪特异性的能量代谢提供了新的思路．     

Effects of cold acclimation on the activity of lipoprotein lipase in adipose tissues of genetically obese Zucker rats

The activity of lipoprotein lipase (LPL) was studied in interscapilar brown adipose tissue (BAT), epididymal white adipose tissue (WAT) and in the heart of lean and obese adult Zucker rats maintained at 22 degrees C or adapted to cold (10 degrees C). In WAT the specific activity per gram of tissue was lower in obese than in lean rats but the total activity within the tissue was three-fold higher. Cold acclimation did not modify total activity in either lean or obese rats. In BAT, but not in the heart, both specific and total activities were lower in obese than in lean animals. They were enhanced in both tissues following cold acclimation. Six-hour fasting led to a decrease in specific activity in WAT of lean rats but had no effect in obese animals; an increase was observed in BAT and heart of both genotypes. Insulin administration has no effect on activities in WAT in either 22 or 10 degrees C adapted obese rats. Norepinephrine administration stimulates LPL activity in BAT and heart of all groups. It is concluded that the lack of development of obesity previously observed in obese rats following cold acclimation is not due to a decreased capacity of lipid uptake by WAT. It might in part be due to an increased lipid oxidation in BAT.     

Cold-induced activation of brown adipose tissue and adipose angiogenesis in mice

Exposure of humans and rodents to cold activates thermogenic activity in brown adipose tissue (BAT). This protocol describes a mouse model to study the activation of BAT and angiogenesis in adipose tissues by cold acclimation. After a 1-week exposure to 4 °C, adult C57BL/6 mice show an obvious transition from subcutaneous white adipose tissue (WAT) into brown-like adipose tissue (BRITE). The BRITE phenotype persists after continuous cold exposure, and by the end of week 5 BRITE contains a high number of uncoupling protein-1-positive mitochondria, a characteristic feature of BAT. During the transition from WAT into BRITE, the vascular density is markedly increased owing to the activation of angiogenesis. In BAT, cold exposure stimulates thermogenesis by increasing the mitochondrial content and metabolic rate. BAT and the increased metabolic rate result in a lean phenotype. This protocol provides an outstanding opportunity to study the molecular mechanisms that control adipose mass.     

The impact of cold acclimation and hibernation on antioxidant defenses in the ground squirrel (Spermophilus citellus): An update

Any alteration in oxidative metabolism is coupled with a corresponding response by an antioxidant defense (AD) in appropriate subcellular compartments. Seasonal hibernators pass through circannual metabolic adaptations that allow them to either maintain euthermy (cold acclimation) or enter winter torpor with body temperature falling to low values. The present study aimed to investigate the corresponding pattern of AD enzyme protein expressions associated with these strategies in the main tissues involved in whole animal energy homeostasis: brown and white adipose tissues (BAT and WAT, respectively), liver, and skeletal muscle. European ground squirrels (Spermophilus citellus) were exposed to low temperature (4 ± 1 °C) and then divided into two groups: (1) animals fell into torpor (hibernating group) and (2) animals stayed active and euthermic for 1, 3, 7, 12, or 21 days (cold-exposed group). We examined the effects of cold acclimation and hibernation on the tissue-dependent protein expression of four enzymes which catalyze the two-step detoxification of superoxide to water: superoxide dismutase 1 and 2 (SOD 1 and 2), catalase (CAT), and glutathione peroxidase (GSH-Px). The results showed that hibernation induced an increase of AD enzyme protein expressions in BAT and skeletal muscle. However, AD enzyme contents in liver were largely unaffected during torpor. Under these conditions, different WAT depots responded by elevating the amounts of specific enzymes, as follows: SOD 1 in retroperitoneal WAT, GSH-Px in gonadal WAT, and CAT in subcutaneous WAT. Similar perturbations of AD enzymes contents were seen in all tissues during cold acclimation, often in a time-dependent manner. It can be concluded that BAT and muscle AD capacity undergo the most dramatic changes during both cold acclimation and hibernation, while liver is relatively unaffected by either condition. Additionally, this study provides a basis for further metabolic study that will illuminate the causes of these tissue-specific AD responses, particularly the novel finding of distinct responses by different WAT depots in hibernators.     

Different metabolic responses of human brown adipose tissue to activation by cold and insulin

We investigated the metabolism of human brown adipose tissue (BAT) in healthy subjects by determining its cold-induced and insulin-stimulated glucose uptake and blood flow (perfusion) using positron emission tomography (PET) combined with computed tomography (CT). Second, we assessed gene expression in human BAT and white adipose tissue (WAT). Glucose uptake was induced 12-fold in BAT by cold, accompanied by doubling of perfusion. We found a positive association between whole-body energy expenditure and BAT perfusion. Insulin enhanced glucose uptake 5-fold in BAT independently of its perfusion, while the effect on WAT was weaker. The gene expression level of insulin-sensitive glucose transporter GLUT4 was also higher in BAT as compared to WAT. In conclusion, BAT appears to be differently activated by insulin and cold; in response to insulin, BAT displays high glucose uptake without increased perfusion, but when activated by cold, it dissipates energy in a perfusion-dependent manner.     

Translocator Protein 18 kDa (TSPO) Is Regulated in White and Brown Adipose Tissue by Obesity

Translocator protein 18 kDa (TSPO) is an outer-mitochondrial membrane transporter which has many functions including participation in the mitochondrial permeability transition pore, regulation of reactive oxygen species (ROS), production of cellular energy, and is the rate-limiting step in the uptake of cholesterol. TSPO expression is dysregulated during disease pathologies involving changes in tissue energy demands such as cancer, and is up-regulated in activated macrophages during the inflammatory response. Obesity is associated with decreased energy expenditure, mitochondrial dysfunction, and chronic low-grade inflammation which collectively contribute to the development of the Metabolic Syndrome. Therefore, we hypothesized that dysregulation of TSPO in adipose tissue may be a feature of disease pathology in obesity. Radioligand binding studies revealed a significant reduction in TSPO ligand binding sites in mitochondrial extracts from both white (WAT) and brown adipose tissue (BAT) in mouse models of obesity (diet-induced and genetic) compared to control animals. We also confirmed a reduction in TSPO gene expression in whole tissue extracts from WAT and BAT. Immunohistochemistry in WAT confirmed TSPO expression in adipocytes but also revealed high-levels of TSPO expression in WAT macrophages in obese animals. No changes in TSPO expression were observed in WAT or BAT after a 17 hour fast or 4 hour cold exposure. Treatment of mice with the TSPO ligand PK11195 resulted in regulation of metabolic genes in WAT. Together, these results suggest a potential role for TSPO in mediating adipose tissue homeostasis.     

Tissue-specific expression of the uncoupling protein family in streptozotocin-induced diabetic rats

The vulnerability of streptozotocin (STZ)-induced diabetic rats to cold stress has been established. One of the elements controlling body temperature is thermogenesis, in which uncoupling protein (UCP) is known to play an important role. We have examined UCP2 and UCP3 expressions in brown adipose tissue (BAT), white adipose tissue (WAT), and skeletal muscle (MSL) during the acute and chronic phases of STZ-induced diabetes in rats. The long-term effect and the effect of insulin treatment thereafter were also unexplored previously and are examined in this study. In the acute phase of diabetes (2.5 days after STZ injection), UCP2 gene expression in BAT, WAT, and MSL, and UCP3 expression in the muscle were significantly increased. In the chronic phase of diabetes (21 days after STZ injection), UCP2 and UCP3 expression in the MSL were restored to the control levels without insulin supplementation. UCP2 in BAT and WAT remained high in the chronic phase, whereas UCP3 expression in BAT and WAT, which did not change in the acute phase, was significantly decreased. Insulin supplementation restored UCP2 expression in BAT and WAT, but over-corrected UCP3 in WAT above the control and did not affect UCP3 expression in BAT. Insulin supplementation depressed UCP3 expression in the MSL below control. These results indicate that the effects of STZ-induced diabetes on UCPs gene expression are tissue-specific as well as dependent on the duration of diabetes.     

禁食及胰岛素水平对大鼠解偶联蛋白-1、2、3基因表达的影响

 为探讨禁食和胰岛素对解偶联蛋白 - 1、2、3基因 (UCP1 ,2 ,3)表达的影响 ,应用 RT- PCR方法观察了在不同禁食时间和应用胰岛素条件下大鼠白色脂肪组织、棕色脂肪组织和骨骼肌中 UCP1 ,2 ,3m RNA水平的变化 .UCP1基因只在大鼠棕色脂肪组织中表达 .UCP2 ,3基因在三种组织中均有表达 ,在白色脂肪组织中以 UCP2表达为主 ;在骨骼肌中以 UCP3表达为主 .过夜禁食使棕色脂肪组织 UCP1 ,3m RNA水平明显下降 (P<0 .0 1 ) ;UCP2 m RNA水平在三种组织中均呈上升反应 ,以白色脂肪组织中表现最为明显 (P<0 .0 5) ;而对白色脂肪组织和骨骼肌中 UCP3基因表达无明显影响 .禁食时间延长至 48h,除棕色脂肪组织中 UCP2 ,3基因有明显下降外 ,各组织中UCPs基因表达基本调节至正常或高于对照组水平 .胰岛素对 UCPs基因表达水平有一定的上调作用 ,这一作用对棕色脂肪组织 UCPs各基因及骨骼肌中 UCP3基因表现得尤为明显 (P<0 .0 5) .大鼠 UCPs基因表达有一定的组织特异性 ;禁食时间对三种组织中 UCPs各成员基因表达的影响有时相上的区别 ;胰岛素可以调 UCPs各成员基因的表达 .结果反映了 UCPs各成员在能量代谢调节上的不同作用 ,这为理解膳食 -产热与体重调节的关系 ,及其能量代谢平衡与疾病关系提供了实验依据