Name: infrared thermography for the detection of changes in brown adipose tissue activity
Uploaded: 2022-09-28T14:30:00
Description: Watch this Scientific Journal Video about Infrared Thermography for the Detection of Changes in Brown Adipose Tissue Activity at JoVE.com

这项研究由克罗地亚科学基金会研究基金（IP-2018-01- 7416）资助。

所有实验动物实验程序均已获得国家伦理委员会和农业部的批准（EP 185/2018）。实验是根据克罗地亚实验动物科学学会的伦理法典和ARRIVE指南进行的。在涉及人类受试者的研究中执行的所有程序均符合赫尔辛基宣言，并经萨格勒布大学医学院伦理委员会批准（UP/I-322-01/18-01/56）。在这项研究中，我们展示了三位女性参与者的结果（BMI：29 kg/m 2 ± 5 kg/m2）。获得所有人类志愿者的知情...

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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+height+explain+gender+differences+in+the+response+to+the+oral+glucose+tolerance+test-the+AusDiab+study.">Differences in height explain gender differences in the response to the oral glucose tolerance test-the AusDiab study.</a> Diabetic Medicine. 25 (3), 296-302 (2008).</li><li>van Genugten, R. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+sex+and+hormone+replacement+therapy+use+on+the+prevalence+of+isolated+impaired+fasting+glucose+and+isolated+impaired+glucose+tolerance+in+subjects+with+a+family+history+of+type+2+diabetes.">Effects of sex and hormone replacement therapy use on the prevalence of isolated impaired fasting glucose and isolated impaired glucose tolerance in subjects with a family history of type 2 diabetes.</a> Diabetes. 55 (12), 3529-3535 (2006).</li><li>Williams, J. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+differences+in+the+prevalence+of+impaired+fasting+glycaemia+and+impaired+glucose+tolerance+in+Mauritius.+Does+sex+matter.">Gender differences in the prevalence of impaired fasting glycaemia and impaired glucose tolerance in Mauritius. Does sex matter.</a> Diabetic Medicine. 20 (11), 915-920 (2003).</li><li>Stanford, K. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brown+adipose+tissue+regulates+glucose+homeostasis+and+insulin+sensitivity.">Brown adipose tissue regulates glucose homeostasis and insulin sensitivity.</a> Journal of Clinical Investigation. 123 (1), 215-223 (2013).</li><li>Brasil, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+on+the+role+of+infrared+thermography+in+the+brown+adipose+tissue+assessment.">A systematic review on the role of infrared thermography in the brown adipose tissue assessment.</a> Reviews in Endocrine and Metabolic Disorders. 21 (1), 37-44 (2020).</li><li>Byers, S. L., Wiles, M. V., Dunn, S. L., Taft, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+estrous+cycle+identification+tool+and+images.">Mouse estrous cycle identification tool and images.</a> PLoS One. 7 (4), 35538 (2012).</li><li>Crane, J. D., Mottillo, E. P., Farncombe, T. H., Morrison, K. M., Steinberg, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+standardized+infrared+imaging+technique+that+specifically+detects+UCP1-mediated+thermogenesis+in+vivo.">A standardized infrared imaging technique that specifically detects UCP1-mediated thermogenesis in vivo.</a> Molecular Metabolism. 3 (4), 490-494 (2014).</li><li>Hartwig, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimodal+imaging+for+the+detection+of+brown+adipose+tissue+activation+in+women:+A+pilot+study+using+NIRS+and+infrared+thermography.">Multimodal imaging for the detection of brown adipose tissue activation in women: A pilot study using NIRS and infrared thermography.</a> Journal of Healthcare Engineering. 2017, 5986452 (2017).</li><li>James, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+infrared+thermography+in+the+measurement+and+characterization+of+brown+adipose+tissue+activation.">The use of infrared thermography in the measurement and characterization of brown adipose tissue activation.</a> Temperature. 5 (2), 147-161 (2018).</li><li>Folgueira, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+dopamine+signaling+regulates+brown+fat+thermogenesis.">Hypothalamic dopamine signaling regulates brown fat thermogenesis.</a> Nature Metabolism. 1 (8), 811-829 (2019).</li><li>Ratko, M., Habek, N., Kordi&#263;, M., Dugand&#382;i&#263;, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+infrared+technology+as+a+novel+approach+for+studies+with+female+laboratory+animals.">The use of infrared technology as a novel approach for studies with female laboratory animals.</a> Croatian Medical Journal. 61 (4), 346-353 (2020).</li></ol>

最近的研究提供了越来越多的证据表明，成年人类和动物的生理调节和BAT活性在肥胖和糖尿病发展中的重要性。此外，外源性激活剂可能激活的BAT正在成为制药公司的目标。为了能够估计BAT在非常繁重的疾病中的生理调节和病理生理重要性，以及发现潜在的治疗方法，红外热成像正在成为首选方法。尽管红外技术正在成为测量BAT活性12，13，14，...

与白色脂肪组织（WAT）相比，棕色脂肪组织（BAT）不储存而是消耗能量。在交感神经刺激下，BAT利用脂肪酸和葡萄糖，并通过激活解偶联蛋白1（UCP1）产生热量。UCP1的功能是利用两个线粒体膜之间的H+梯度来产生热量，而不是ATP。BAT的功能是在寒冷条件下增加热量产生，从而导致能量消耗增加1。冷暴露后，来自皮肤的感觉输入抑制下丘脑视前区 （POA） 正中视前 （MnPO） 核中的热敏感神经元，从而降低 POA 神经元对苍白嘴 （rRPa） 的抑制作用。rRPa神经元的激活增加交感神经活性，随后BAT活性增加2，3。冷诱导的BAT激活可改善人类的胰岛素敏感性4，并且在体重指数（BMI）增加和年龄为1，5，6，7的人类中，这种活性降低。
除了在冷诱导产热中的作用外，饭后，英蝙蝠对瘦男性人群的葡萄糖摄取增加，有助于饮食诱导的产热（DIT），这在BAT阳性男性受试者中更高8，9。用于测量BAT活性的最先进技术是正电子发射断层扫描计算机断层扫描，称为PET-CT。该方法通过测量放射性示踪剂氟脱氧葡萄糖（18F-FDG）的积累来确定BAT活性。然而，PET-CT作为检测餐后BAT激活的首选方法失败。原因之一是，餐后，18F-FDG与餐后高血糖竞争相同的葡萄糖转运蛋白，这使得它不适合确定餐后BAT激活，特别是在比较健康和糖尿病参与者的BAT活性时血糖浓度可能存在差异。此外，BAT使用脂肪酸作为热量产生的能量来源，这是PET-CT所不可见的。18餐后蝙蝠中的F-FDG积累几乎看不见10，因此在大多数情况下被解释为阴性结果。不出所料，最近有人提出，BAT的激活在人群中比我们以前想象的更明显;因此，有必要采用一种新的方法来检测BAT活性及其参与代谢紊乱7。解决这个问题的尝试是用磁共振成像（MRI）测量糖尿病前期患者和具有胰岛素抵抗的2型糖尿病（T2DM）患者的BAT体积11。然而，通过MRI测量的BAT体积不足以估计BAT的日常功能和葡萄糖和脂肪酸的使用。因此，为了估计健康与T2DM患者BAT活性的实际差异，需要一种新的方法，为找出T2DM患者BAT功能障碍的病理机制提供可能性。
为了确定BAT的激活，我们使用红外（IR）热成像技术测量了餐前和饭后的BAT热量产生（图1）12，13。将红外热成像技术作为测量健康和肥胖个体或糖尿病患者餐后BAT活性的首选方法，将对现场产生巨大影响。时至今日，红外热成像仍用于测定 BAT13、14、15 的冷诱导活化。在近代人类历史上，寒冷引起的BAT活动不再非常明显（由于栖息地的适当加热，适当的衣服），而饭后BAT激活每天都在发生。此外，这两种BAT功能通过下丘脑的生理调节是完全不同的。饭后，下丘脑弓形核（Arc）中表达前黑皮质素（POMC）的神经元的激活导致交感神经活动通过rRPa16增加。通过红外热成像或PET-CT测量的冷诱导的BAT活化在用作日常BAT活动的测量时是不合适的。餐后BAT活性增加之后是葡萄糖利用，这对于维持葡萄糖稳态，胰岛素敏感性和葡萄糖浓度的日常调节最终很重要。餐后BAT激活导致餐后葡萄糖消耗增加，随后热量产生和体温（DIT）增加。这被证明是性别，年龄和BMI依赖12。在雄性和雌性实验室小鼠中观察到餐后BAT激活的相似性别差异17。这些发现对应于Burke等人最近发现的BAT调节的性别差异，他们表明通过POMC神经元亚群对BAT褐变的下丘脑调节在雄性和雌性小鼠中有所不同18。女性、老年人群和肥胖人群中 BAT 的餐后激活较小。餐后缺乏BAT激活（葡萄糖利用率降低）可能导致女性糖耐量受损的患病率更高19，20，21，22。不幸的是，大多数关于BAT激活的研究只在男性身上进行。通过在饭后激活BAT，瘦男性人群的葡萄糖摄取增加。毫不奇怪，在BAT激活后，BAT阳性男性受试者的DIT更高8，9。此外，雄性小鼠的BAT移植改善了葡萄糖耐量，增加了胰岛素敏感性，并降低了体重和脂肪量23。
PET-CT作为测量BAT活性的首选方法失败，尤其是在饭后。因此，开发了一种非侵入性和更敏感的方法。红外热成像能够估计不同实验动物（敲除小鼠）以及人类参与者的BAT活性，无论性别，年龄或不同病理条件对BAT活性的影响如何。这种方法的另一个好处是参与者和实验动物的简单性，这使我们能够估计BAT加强治疗的潜在益处。最近使用红外热成像来确定寒冷暴露或用餐后BAT生理行为的研究在最近发表的Brasil等人24中有所描述。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.1% cresyl violet acetate&#160;</td>
			<td>Commonly used chemical</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Device for measuring air temperature and humidity</td>
			<td>Kesterl</td>
			<td>Kestrel 4200</td>
			<td>Certificat of conformity</td>
		</tr>
		<tr>
			<td>External data storage</td>
			<td>Hard Drive with at least 1 TB</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glass microscopic slides</td>
			<td>Commonly used</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Small cotton tip swab&#160;</td>
			<td>Urethral swabs</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Software for analysis</td>
			<td>FLIR Systems, Wilsonville, OR, USA</td>
			<td>FLIR Tools</td>
			<td></td>
		</tr>
		<tr>
			<td>Software for meassurements</td>
			<td>FLIR Systems, Wilsonville, OR, USA</td>
			<td>ResearchIR software</td>
			<td>FLIR ResearchIR Max, version 4.40.12.38 (64-bit)</td>
		</tr>
		<tr>
			<td>Thermac Camera</td>
			<td>FLIR Systems, Wilsonville, OR, USA</td>
			<td>FLIR T-1020</td>
			<td></td>
		</tr>
	</tbody>
</table>

infrared thermography for the detection of changes in brown adipose tissue activity

通过正电子发射断层扫描计算机断层扫描（PET-CT）通过餐后或肥胖或糖尿病患者积累的18F-氟脱氧葡萄糖（FDG）来测量棕色脂肪组织（BAT）活性，作为首选方法失败。主要原因是18F-FDG与餐后高血糖血浆浓度竞争BAT细胞膜上相同的葡萄糖转运蛋白。此外，BAT也使用脂肪酸作为能量来源，这在PET-CT中是不可见的，并且可以随着肥胖和糖尿病患者的葡萄糖浓度而改变。因此，为了估计BAT在动物和人类中的生理重要性，应用了最近出版物中使用的一种新的红外热成像方法。
禁食过夜后，通过人类志愿者和雌性野生型小鼠餐前和饭后的红外热成像测量BAT活性。相机软件使用与物体的距离、皮肤发射率、反射室温、空气温度和相对湿度来计算物体的温度。在小鼠中，BAT上方的剃光区域是测量平均和最大温度的感兴趣区域。雌性小鼠的发情周期阶段是在用甲酚紫（0.1%）染色溶液染色的阴道涂片实验后确定的。在健康志愿者中，选择了颈部的两个皮肤区域：锁骨上区域（锁骨上方，存在BAT细胞）和锁骨间区域（锁骨之间，没有检测到BAT组织）。BAT 活动由这两个值的减法确定。此外，可以在动物和人类参与者中确定皮肤区域的平均和最大温度。
通过红外热成像（一种非侵入性和更灵敏的方法）测量的餐后BAT活性的变化被证明是实验动物的性别，年龄和发情周期的阶段。作为饮食诱导的产热的一部分，人类的BAT激活也被证明与性别，年龄和体重指数有关。进一步确定餐后BAT活性的病理生理变化对于高葡萄糖血浆浓度（肥胖和糖尿病2型）以及不同实验动物（敲除小鼠）的参与者非常重要。这种方法也是确定可能激活BAT活性的药物的可变工具。

确定BAT活性的最简单方法是减去人类受试者饭前和饭后高于BAT的最大皮肤温度。计算BAT活性的更好方法是选择两个感兴趣的区域：BAT上方的皮肤区域，位于锁骨上区域，以及皮肤锁骨间区域，其中人类没有发现BAT组织，指定为参考区域（根据PET-CT;图1）。然后通过减去这两个温度很容易确定BAT活性。如图1所示，确定BAT活性为1.8°C。 为了确定喂食后BAT的...

Watch this Scientific Journal Video about Infrared Thermography for the Detection of Changes in Brown Adipose Tissue Activity at JoVE.com

Infrared Thermography for the Detection of Changes in Brown Adipose Tissue Activity

在这里，我们提出了一种测量人类和实验动物餐后棕色脂肪组织活性的方案。

红外热成像用于检测棕色脂肪组织活性的变化

Measuring brown adipose tissue (BAT) activity by positron emission tomography computed tomography (PET-CT) via the accumulation of 18F-fluorodeoxyglucose ...

红外热成像能够估计实验动物和人类参与者的棕色脂肪组织活性，无论性别、年龄或不同病理状况的影响如何。PET-CT作为测量棕色脂肪组织活性的首选方法失败，尤其是在饭后。因此，开发了一种无创且更灵敏的红外热成像技术。利用红外热成像，我们可以确定肥胖和糖尿病患者棕色脂肪组织活性或沐浴活性的病理生理变化，并确定可以使这些患者的不良活动恢复活力的药物。该方法可用于确定激素或合成治疗物质对人类和实验动物不良活性的生理作用。询问参与者是否休息良好并禁食过夜。他们将在测量前休息至少 30 分钟，以避免可能的棕色脂肪组织激活。在早上进行实验。测量前15分钟，要求参与者脱掉上衣。当参与者休息时，将热像仪安装在三脚架上，并将其放置在距离参与者坐的位置一米远的地方。按照制造商的说明将热成像摄像机连接到计算机和软件。通过在一米的焦距处记录起皱的铝箔来确定房间的反射温度。为此，在相机软件中，输入零米的距离和 1 的发射率。然后，输入大气压力和相对湿度的值。要确定反射温度，请选择正方形并选择铝箔上的ROI。然后，在主相机窗口中选择左侧的第一个图标并读取平均温度，这是进一步分析所必需的。在开始测量之前，确定室内空气温度和空气湿度。在软件中，在主相机窗口上方，选择左侧的第三个图标。在弹出菜单中，选取“编辑记录设置”，这将打开一个新窗口。在“录制”模式下，选择“记录到磁盘”，然后在所需时间设置此持续时间的记录。在同一窗口的“录制选项”中，将录制速率限制为 5 赫兹，然后选择保存录制的位置。关闭现有窗口，在主菜单中打开编辑，然后选择首选项。在目标帧速率中输入 5。选择热键/远程启动可以停止记录。然后从下拉菜单中选择启动/停止模式。在测量时，确保只有参与者和实验者在场。避免空气流动，并确保参与者远离冷风、阳光或任何热源，如灯泡。将颈部锁骨上区域定位在锁骨上方，棕色脂肪组织所在的位置。将参与者定位为该区域，使其焦距为一米。按 F5 键以 5 FPS 的帧速率录制短片 10 到 15 秒。确保所有参与者都吃同一顿饭。饭后，在 30 分钟进行新录制，然后使用相同的设置值按 F5 进行 1、2 和 3 小时。在实验前一天，收集动物的阴道拭子并将细胞散布在载玻片上。让它风干。用500微升0.1%甲酚紫醋酸酯染色细胞一分钟，然后用水冲洗三次。在具有100倍放大倍率和明场照明的光学显微镜下观察细胞。根据涂片中观察到的白细胞数量以及有核和定量的上皮细胞确定发情周期的阶段。在实验的早晨，准备热像仪和记录设置。小心地将动物放在干净的笼子里，并将笼子放在热像仪下方，焦距为一米。按 F5 录制动画。在给每只动物之前称量食物颗粒以计算食物摄入量。让动物在笼子里吃 30 分钟，饭后再次称量食物颗粒。开始用餐后 30 分钟、1 小时和 2 小时重复 IR 测量。实验结束后，测试雌性动物发情周期的阶段，如前所述。对于每部电影，在相机软件中输入变量，例如皮肤发射率、反射室温、空气温度、相对湿度和与物体的距离。通过移动屏幕底部的播放头或按下暂停按钮，从影片中选择合适的帧。通过在主窗口左侧选择所需形状的区域来选择感兴趣的区域。选择最符合肩胛骨之间皮肤区域的形状。选择感兴趣区域后，最小、最高和平均温度将显示在右侧。在图像中，红色三角形表示最高记录温度点，蓝色三角形表示最低记录温度。重复此步骤几帧，以确保测量的温度在录制的几秒钟内保持稳定。从餐后的最高温度中减去餐前棕色脂肪组织上方皮肤的最高温度，以确定餐后活动的增加。通过分析两个皮肤区域来确定棕色脂肪组织活性：一个在棕色脂肪组织上方，第二个在锁骨间区域作为参考点。棕色脂肪组织活性根据两个皮肤区域的最高温度差异计算为1.8摄氏度。但是这种活动不能通过棕色脂肪组织上方皮肤的最大温度来呈现，因为即使在进餐三小时后皮肤温度也没有降低。在小鼠中，通过测量棕色脂肪组织上方皮肤的平均温度来测量雌性餐后活动的变化。只有在测量最高温度时，饭后30分钟才确定活性的显着变化。正确选择匹配的对照受试者是临床研究最困难的部分，因为健康对照和有病理状况的参与者应尽可能相似，并且仅在所调查的疾病上有所不同。除了该协议之外，还可以测量血糖浓度和体温，这允许对不良活动与葡萄糖利用或体温升高进行相关性分析。这种敏感和非侵入性技术为确定其他实验环境和研究中棕色脂肪组织活性的生理和病理生理变化提供了许多可能性。

Research

JoVE Journal

Medicine

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

科研

医学

Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery

组织鬼怪模拟，以评估乳癌手术潜在的近红外荧光成像中的应用

Inaccuracies in intraoperative tumor localization and evaluation of surgical margin status result in suboptimal outcome of breast-conserving surgery ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

肩胛间褐色脂肪组织的切伦科夫发光成像

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Human Brown Adipose Tissue Depots Automatically Segmented by Positron Emission Tomography/Computed Tomography and Registered Magnetic Resonance Images

人类的褐色脂肪组织的仓库的正电子发射断层扫描/计算机断层扫描自动分段及注册磁共振图像

Reliably differentiating brown adipose tissue (BAT) from other tissues using a non-invasive imaging method is an important step toward studying BAT in ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

应用 Videomicroscopy 评价人脂肪组织微血管功能

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

自体 Microfractured 与纯化脂肪组织在关节镜下治疗距骨软骨病变

In recent years, regenerative techniques have been increasingly studied and used to treat osteochondral lesions of the talus. In particular, several ...

Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

应用流式细胞仪对人体脂肪组织免疫细胞的表征

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

危重病人全血内毒素血症检测的内毒素活性测定

Lipopolysaccharide, also known as endotoxin, is a fundamental component of gram-negative bacteria and plays a crucial role in the development of sepsis ...