Name: effects of exposure of formaldehyde to a rat model of atopic dermatitis induced by neonatal capsaicin treatment
Uploaded: 2017-09-27T12:30:00.000+00:00
Duration: 6 min 47 s
Description: Watch this Scientific Journal Video about Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment at JoVE.com

这项研究得到了公共福利和 #38 的支持; 通过由教育、科学和技术部 (NRF-2014M3C8A5030612) 资助的韩国国家研究基金会 (NRF) 的安全研究项目, 并得到了中杨、ChaSun, 医学博士和宇 Sa 纪念基金。

此处描述的所有方法均已获得韩国大学医学院国际动物护理与使用委员会 (IACUC) 的批准. #160; 出生后6周, 实验者可诱发 enthansasia 和 #160; 收集各种样本, 如大鼠的皮肤和血清. 
 1. 新生儿辣椒素治疗 AD 的诱导 <ol> <li> 要准备5毫克/毫升的辣椒素溶液, 在80毫升锥形管中加入50毫克的辣椒素到1毫升的吐温1和100% 毫升的15乙醇. </li>
	<li> 添加8毫升生理盐水到锥形管。用力摇动管, 直到没有分离的图层可见 </li> <li> 将辣椒素溶液 (10-#181; L/g) 注入到皮下组织, 在新生儿大鼠颈部中线和背部。
	<ol> <li> 在出生后12ĥ至48小时之间提供辣椒素 (关键). 
		注: 如果注射延迟超过几天, 皮炎将不会发展, 或只会持续几个星期。如果注射过早, 会因剧烈疼痛而发生呼吸窘迫. </li>
		<li> 为了防止新生鼠因辣椒素注射液引起的呼吸道骤停, 推动新生儿的胸部, 以鼓励呼吸或保持新生儿在氧气室, 直到呼吸正常. 和 #160, 因为辣椒素引起的疼痛和 #160;劝阻呼吸, 增加全身麻醉和 #160; #160;p ossibly 会使呼吸困难恶化。因此, 实验者不应该使用 anesthesics 来增加老鼠的存活率. </li>
	</ol> </li> </ol> 2。AD 大鼠皮肤瘙痒和皮炎的评价 <ol> <li> 计数自发划伤行为的瘙痒测量 <ol> <li> 准备塑料室 (20 cm x 30 cm x 20 cm), 配有一面镜子, 以允许全面覆盖查看和孔, 让老鼠呼吸. </li>
		<li> 正确设置和调整数字摄像机, 以便同时记录大鼠的镜像视图和前视图. </li>
		<li> 将老鼠放在塑料室里, 用重物遮住房间的天花板, 以防打开. </li>
		<li> 记录1小时的大鼠自发行为, 播放视频片段, 并计算划伤行为的次数. </li>
	</ol> </li> <li> 使用评分系统评估皮肤损害 <ol> 将老鼠置于感应室. <li>
		<li> 将氧气流量计调至2升/分钟流量, 并将异氟醚蒸发器调整为3% 进行麻醉. </li>
		<li> 识别大鼠耳部和其他部位皮炎的程度和严重性. </li>
		<li> 定义皮肤损害程度为 0.25 cm 2 的单位大小和皮肤病变的严重程度, 按以下 表 1 : 严重度指数; 皮炎评分乘以程度和严重性 (见皮炎评分在强类 = "xfig" > 图 1B ). </li>
	</ol> </li> </ol> 3。气体 FA 对动物的暴露 <ol> <li> 吸入室的制备 <ol> <li> 准备一个亚克力玻璃盒 (55 厘米 x 40 cm x 35 厘米), 它的两侧有两个插座. </li>
		<li> 警告。对于 FA 气体的循环, 将出水口放在高于进口的位置, 使出水口的尺寸大于进口. </li>
		<li> 警告。对于内部气体浓度的测量, 在出口的另一侧, 钻三小通道, 可以关闭. </li>
		<li> 将管道连接到两个插座。将排气管放在建筑物的通风罩或外侧, 以排放气体, 并将其它出口管连接至医用氧气调节器. </li>
		<li> 在亚克力玻璃盒内放置一个传统的鼠笼. </li>
		<li> 在出生后一周内将老鼠放在笼子里5周 (2 小时/天, 5 天/周). </li>
		<li> 尽可能紧密地密封亚克力玻璃盒以防止漏气. 
		注: 密闭密封时, 箱盖应含橡胶。用保鲜膜包住密封腔. </li>
	</ol> </li> <li> 使用医用氧气调节器生成气体 fa <ol> 将5克 fa 溶解为1升水. <li>
		<li> 将 0.5% FA 溶液放在医用氧气调节器的加湿器瓶中. </li>
		<li> 将入口管连接到调节器后, 打开调节器并将流量设置为5升/分. </li>
		<li> 调整溶液浓度和流速以确定燃烧室内气体的浓度. </li>
	</ol>
	</li>
</ol>

<ol>
	<li>Geha, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allergy+and+hypersensitivity.+Nature+versus+nurture+in+allergy+and+hypersensitivity.">Allergy and hypersensitivity. Nature versus nurture in allergy and hypersensitivity.</a> Curr Opin Immunol. 15 (6), 603-608 (2003).</li><li>Novak, N., Bieber, T., Leung, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+mechanisms+leading+to+atopic+dermatitis.">Immune mechanisms leading to atopic dermatitis.</a> The J Allergy Clin Immunol. 112 (Suppl 6), S128-S139 (2003).</li><li>Back, S. K., 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=Chronically+relapsing+pruritic+dermatitis+in+the+rats+treated+as+neonate+with+capsaicin;+a+potential+rat+model+of+human+atopic+dermatitis.">Chronically relapsing pruritic dermatitis in the rats treated as neonate with capsaicin; a potential rat model of human atopic dermatitis.</a> J Dermatol Sci. 67 (2), 111-119 (2012).</li><li>Jin, H., He, R., Oyoshi, M., Geha, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+atopic+dermatitis.">Animal models of atopic dermatitis.</a> J Invest Dermatol. 129 (1), 31-40 (2009).</li><li>Han, R. T., 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=Formaldehyde-Induced+Aggravation+of+Pruritus+and+Dermatitis+Is+Associated+with+the+Elevated+Expression+of+Th1+Cytokines+in+a+Rat+Model+of+Atopic+Dermatitis.">Formaldehyde-Induced Aggravation of Pruritus and Dermatitis Is Associated with the Elevated Expression of Th1 Cytokines in a Rat Model of Atopic Dermatitis.</a> PLOS ONE. 11 (12), e0168466 (2016).</li><li>Xu, B., Aoyama, K., Takeuchi, M., Matsushita, T., Takeuchi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+cytokine+mRNAs+in+mice+cutaneously+exposed+to+formaldehyde.">Expression of cytokine mRNAs in mice cutaneously exposed to formaldehyde.</a> Immunol Lett. 84 (1), 49-55 (2002).</li><li>Kim, J. Y., Jeong, M. S., Park, K. Y., Seo, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aggravation+of+atopic+dermatitis-like+symptoms+by+consecutive+low+concentration+of+formaldehyde+exposure+in+NC/Nga+mice.">Aggravation of atopic dermatitis-like symptoms by consecutive low concentration of formaldehyde exposure in NC/Nga mice.</a> Exp Dermatol. 22 (3), 219-221 (2013).</li><li>Laske, N., Niggemann, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+the+severity+of+atopic+dermatitis+correlate+with+serum+IgE+levels?.">Does the severity of atopic dermatitis correlate with serum IgE levels?.</a> Pediatr Allergy Immunol. 15 (1), 86-88 (2004).</li><li>Fujii, Y., Takeuchi, H., Sakuma, S., Sengoku, T., Takakura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+2,4-dinitrochlorobenzene-induced+chronic+dermatitis+model+in+rats.">Characterization of a 2,4-dinitrochlorobenzene-induced chronic dermatitis model in rats.</a> Skin Pharmacol Physiol. 22 (5), 240-247 (2009).</li><li>Sugimoto, T., Xiao, C., Ichikawa, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+primary+neuronal+death+induced+by+capsaicin+and+axotomy+involves+an+apoptotic+mechanism.">Neonatal primary neuronal death induced by capsaicin and axotomy involves an apoptotic mechanism.</a> Brain Res. 807 (1-2), 147-154 (1998).</li><li>Saito, 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=Characterization+of+skin+inflammation+induced+by+repeated+exposure+of+toluene,+xylene,+and+formaldehyde+in+mice.">Characterization of skin inflammation induced by repeated exposure of toluene, xylene, and formaldehyde in mice.</a> Environ Toxicol. 26 (3), 224-232 (2011).</li><li>Jung, W. 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=Formaldehyde+exposure+induces+airway+inflammation+by+increasing+eosinophil+infiltrations+through+the+regulation+of+reactive+oxygen+species+production.">Formaldehyde exposure induces airway inflammation by increasing eosinophil infiltrations through the regulation of reactive oxygen species production.</a> Environ Toixcol Pharmacol. 24 (2), 174-182 (2007).</li></ol>

作者没有什么可透露的。

本文所描述的用于诱导大鼠 ad 样症状的协议可用于研究 ad 病理生理学, 并作为药物测试的筛选工具。该仪器的气体 FA 接触辣椒素治疗大鼠可用于评估的影响, 接触各种气体的不同动物模型。
新生儿辣椒素治疗 AD 模型依赖于比其他特应性模型更简单的过程, 如 DNCB 治疗模型9和 epicutaneous 敏化模型4, 并且比 NC/衙鼠标4更经济实惠。ad 样症状的表现在广告模型中是一致的。此外, 在某些情况下, 该协议会引起受影响的大鼠自发复发的 AD 样皮肤病变, 而不需要任何额外治疗3。过量的辣椒素注射液诱发神经细胞死亡和神经营养炎症10 , 并引起大鼠的严重疼痛。他们中的一些人可能因疼痛而患上呼吸骤停。这是很重要的, 以防止他们呼吸停止。因此, 辣椒素注射液后, 幼崽应放置在氧气室和一些幼崽可能需要胸部压缩。新生儿辣椒素治疗不会诱发小鼠的 AD 样症状, 原因尚不清楚。因此, 基因干预是很难适用于这个 AD 模型使用了淘汰赛的应变, 但其他的遗传方法, 如 siRNA 振振有词地弥补这一缺陷。此外, 大鼠模型的 ad 样症状的一致表现有利于研究 ad 疾病的发病机制, 并将其应用于药物筛选工具。
用于 FA 接触的循环室的建立只需要传统的实验室设备, 包括一个亚克力盒和一个医用氧气调节器。此外, 通过改变氧气调节器加湿器瓶中的溶液, 可以在实验中应用不同类型的气体。通过调节调节器的流速和溶液的浓度, 可以很容易地保持燃烧室内气体的浓度。这是至关重要的, 以确保在安装的循环系统, 为 FA 接触辣椒素治疗 AD 大鼠模型使用丙烯酸室和医疗氧调节器。不仅因为室内的气体浓度会因泄漏而不同, 而且泄漏的气体也会对实验者造成伤害。某些挥发性有机化合物 (VOC) 的汽化通常会通过结晶来阻挡管子。因此, 它是相关的, 以避免堵塞通过连续管管理长期使用, 定期更换管。这种汽化系统比局部应用方法11更准确地反映了患者的疾病进展。这一方法揭示了 fa 吸入对大鼠 AD 模型的明显影响, 与以往暴露于幼稚动物的研究相比,7,12。

为了探讨甲醛 (fa) 吸入对特应性皮炎 (ad) 进展的影响, 我们建立了一个用于 ad 诱导的协议, 并开发了一种用于慢性 fa 的常规循环室。
AD 是一种慢性炎症性皮肤病, 归因于遗传易感性与环境触发的复杂相互作用1,2。需要一个合适的动物模型来研究 AD 的临床前状态。在环境因素的影响下阐明 AD 的发病机制是最重要的。此外, 为了更好地研究环境诱因对广告进展的影响, 必须区分个体环境因素和量化暴露程度。
我们展示了一种新的大鼠 AD 诱导的新生儿辣椒素治疗模型。此 ad 模型3与其他 ad 模型 (如 epicutaneous 敏化、基因工程 ad 模型以及 ad4的自发鼠标模型) 所引起的 ad 模型具有某些优点。首先, 该模型在3周的时候开发出类似于 ad 的症状, 与婴儿 ad 相对应, 并且比其他自发的小鼠模型 (如 NC/衙鼠标) 更早。在某些情况下, 与基因工程广告模型不同, ad 模型的症状会在20周内自发消退和复发。此外, 模型中的 AD 样症状的诱导过程比 epicutaneous 敏化模型容易, 这与诱导过程中的过敏性接触性皮炎更相似。
我们通过使用丙烯酸玻璃腔和传统的医用氧调节器5 , 介绍了 fa 接触动物模型的新方法, 研究了 fa 吸入对婴幼儿期 AD 发育的影响。调节器允许我们通过调节流速和溶液浓度来使动物接触到恒定的气体浓度。以前的研究已经报告了几种方法暴露 FA 动物模型。徐et al.描述了 FA 解决方案直接应用于皮肤6的曝光方法。另一项最近的研究讨论了使用专门的挥发性溶剂发生器, 模型 49127的 FA 气体暴露的协议。与以往的研究方法相比, 该方法更易于配置, 除 FA 外, 还可应用于其他解决方案。

<table><tbody><tr><td>Medical Oxygen Regulator</td><td>Yuyao Jiahua Medical</td><td></td><td>manufacturer will not affect results</td></tr><tr><td>Tube</td><td></td><td></td><td>manufacturer will not affect results</td></tr><tr><td>Acrylic Chamber</td><td>Taejin Acryl</td><td></td><td>manufacturer will not affect results</td></tr><tr><td>Plastic Chamber</td><td>Taejin Acryl</td><td></td><td>manufacturer will not affect results</td></tr><tr><td>Paraformaldehyde, powder</td><td>Sigma</td><td>158127</td><td>manufacturer will not affect results</td></tr><tr><td>Wrap filme</td><td>Uniwrap</td><td></td><td>manufacturer will not affect results</td></tr><tr><td>Capsaicin</td><td>Sigma</td><td>M2028</td><td>manufacturer will not affect results</td></tr><tr><td>Tween 80</td><td>Sigma</td><td>P4780</td><td>manufacturer will not affect results</td></tr><tr><td>Digital Video Camera</td><td>Sony</td><td>HDR-CX380y</td><td>manufacturer will not affect results</td></tr><tr><td>Isoflurane</td><td>JW pharmaceutical</td><td></td><td>manufacturer will not affect results</td></tr></tbody></table>

effects of exposure of formaldehyde to a rat model of atopic dermatitis induced by neonatal capsaicin treatment

特应性皮炎长期复发瘙痒湿疹和盛行于世界各地, 特别是在发达国家。遗传和环境因素之间的复杂相互作用已知在特应性皮炎的病理生理学中起重要作用。然而, 我们仍然缺乏这一疾病的发病机制的详细情况。因此, 建立适当的动物模型来阐明特应性皮炎的进展是非常重要的。此外, 调查空气污染物等环境因素对特应性皮炎的影响, 扩大了对该病的认识。在这里, 我们描述了一种诱导新生辣椒素治疗大鼠特应性皮炎的方法, 以及一项将甲醛浓度恒定暴露在大鼠体内以揭示其对婴幼儿特应性皮炎发展影响的协议时期.这些协议已成功地应用于一些实验, 并可用于其他物质。

7周龄的辣椒素治疗 ad 大鼠模型的 ad 样皮肤炎症在图 1A中介绍。在视频 1中显示了辣椒素治疗的 AD 大鼠模型的抗刮伤行为。新生儿辣椒素治疗引起皮炎和严重抓伤行为的大鼠3周龄, 和 AD 样症状持续了几个星期 (图 1B, C)。虽然程度不那么严重, 一些大鼠遭受复发性皮炎在16周的年龄后, 早期的广告样症状的解决 (图 1D)。此外, 血清 IgE 水平是 ad 严重程度8的生物标记, 在辣椒素治疗的 ad 鼠模型 (图 1E) 中升高。IL-4 和 IL-13 的 mRNA 表达也在辣椒素治疗的 AD 大鼠模型 (图 1F, G) 中增加。
有代表性的照片显示, FA 在5周龄的辣椒素治疗的 AD 大鼠模型中诱发皮炎的加重 (图 2A)。1.2 ppm 法暴露在辣椒素治疗的 AD 大鼠模型中的抓挠行为和皮炎超过2周。然而, 0.8 ppm FA 暴露没有加剧抓挠行为和皮炎的辣椒素治疗 AD 大鼠模型 (图 2B, C)。此外, 1.2 ppm FA 暴露提高了血清 IgE 水平的辣椒素治疗 AD 大鼠模型 (图 2D)。
对于气体 FA 的曝光, 通风室是按照协议(图 3A) 中的描述设置的。暴露前, 在室内的 FA 浓度反复测量, 以保证浓度一致。在改变 fa 溶液浓度的同时, 测定了气相法在室内的浓度。随着溶液浓度的增加, 气体在燃烧室中的浓度逐渐增加, 而同一溶液中的气体浓度在重复测量中没有明显变化 (图 3B)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55987/55987fig1.jpg"> 
图 1: 新生辣椒素治疗诱发复发性 AD 样皮肤炎症和瘙痒的大鼠.辣椒素治疗大鼠皮肤炎症的代表性照片 (A)。辣椒素治疗大鼠皮炎 (B) 和抓挠行为 (C) 的年代学特征。16周老辣椒素治疗大鼠复发性皮肤病变的代表性图像 (D)。(E) 在辣椒素治疗的动物中血清 IgE 水平显著升高。(F) 在辣椒素治疗的动物中 IL-4 mRNA 表达的显著升高。(G) 在辣椒素治疗的动物中 IL-3 mRNA 表达的显著升高。所有结果均以 x 扫描电镜和学生 t 检验法进行分析。*p & #60; 0.05、*** 和 #60; 0.005, **p 和 #60; 0.001。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55987/55987fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55987/55987fig2.jpg"> 
图 2: 在辣椒素治疗的 AD 大鼠模型中暴露严重的瘙痒和皮炎.使用 1.2 ppm 和 0.8 ppm FA (A) 处理的 5th周的代表性照片。暴露 1.2 ppm fa, 但不是 0.8 ppm fa 曝光, 明显加剧瘙痒 (B) 和皮炎 (C) 在辣椒素治疗 AD 模型。1.2 ppm 暴露 (D) 治疗的辣椒素治疗 AD 大鼠模型血清 IgE 水平升高。结果被表达为手段±扫描电镜和分析使用的 one-way 方差检验后, Tukey 的测试或学生的 t 检验。*p & #60; 0.05, **p 和 #60; 0.001 (vs 大鼠暴露在新鲜空气中)。1.2 fa: 辣椒素处理的 ad 大鼠模型暴露于 1.2 ppm fa;0.8 fa: 辣椒素处理的 ad 大鼠模型暴露于 0.8 ppm fa;和空气广告: 辣椒素治疗的 ad 大鼠模型暴露在新鲜空气中。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55987/55987fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55987/55987fig3.jpg"> 
图 3: FA 处理.调节器和气室 (A) 的示意图说明。气态 fa 浓度在燃烧室中与 fa 溶液浓度成正比 (B)。结果被表达为手段± SEM.<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55987/55987fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Video" src="/files/ftp_upload/55987/55987video1.jpg"> 
视频 1: 对辣椒素处理大鼠的抓挠行为进行了记录, 以测量瘙痒.<a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55987/Video_Figure.mp4" target="_blank">请单击此处查看此视频。(右键单击可下载.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>区域</td>
			<td>得分</td>
			<td>描述</td>
		</tr>
		<tr>
			<td rowspan="4">除了耳朵</td>
			<td>0</td>
			<td>正常</td>
		</tr>
		<tr>
			<td>1</td>
			<td>稀疏毛皮</td>
		</tr>
		<tr>
			<td>2</td>
			<td>脱发或闪光</td>
		</tr>
		<tr>
			<td>3</td>
			<td>出血或痂</td>
		</tr>
		<tr>
			<td rowspan="4">耳朵</td>
			<td>0</td>
			<td>正常</td>
		</tr>
		<tr>
			<td>1</td>
			<td>flare</td>
		</tr>
		<tr>
			<td>2</td>
			<td>出血或痂</td>
		</tr>
		<tr>
			<td>3</td>
			<td>组织的损失</td>
		</tr>
	</tbody>
</table>
表 1:严重性指数

Watch this Scientific Journal Video about Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment at JoVE.com

Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment

我们在此描述了新生儿辣椒素治疗诱导大鼠特应性皮炎的方法, 并探讨了甲醛吸入对过敏性皮炎的影响。

甲醛暴露对大鼠新生辣椒素致过敏性皮炎模型的影响

Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between ...

effects-exposure-formaldehyde-to-rat-model-atopic-dermatitis-induced

Universidad San Sebastian

Research

JoVE Journal

Immunology and Infection

9.6K 次观看.  Korea University. 我们在此描述了新生儿辣椒素治疗诱导大鼠特应性皮炎的方法, 并探讨了甲醛吸入对过敏性皮炎的影响。

视频: Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

科研

行为学

Separation of Rat Epidermis and Dermis with Thermolysin to Detect Site-Specific Inflammatory mRNA and Protein

Easy-to-use and inexpensive techniques are needed to determine the site-specific production of inflammatory mediators and neurotrophins during skin injury, inflammation, and/or sensitization. The goal of this study is to describe an epidermal-dermal separation protocol using thermolysin, a proteinase that is active at 4 &deg;C. To illustrate this procedure, Sprague Dawley rats are anesthetized, and right hind paws are injected with carrageenan. Six and twelve hours after injection, rats with inflammation and na&iuml;ve rats are euthanized, and a piece of hind paw, glabrous skin is placed in cold Dulbecco's Modified Eagle Medium. The epidermis is then separated at the basement membrane from the dermis by thermolysin in PBS with calcium chloride. Next, the dermis is secured by microdissection forceps, and the epidermis is gently teased away. Toluidine blue staining of tissue sections show that the epidermis is separated cleanly from the dermis at the basement membrane. All keratinocyte cell layers remain intact, and the epidermal rete ridges along with indentations from dermal papillae are clearly observed. Qualitative and real-time RT-PCR is used to determine nerve growth factor and interleukin-6 expression levels. Western blotting and immunohistochemistry are finally performed to detect amounts of nerve growth factor. This report illustrates that cold thermolysin digestion is an effective method to separate epidermis from dermis for evaluation of mRNA and protein alterations during inflammation.

Presented here is a protocol for the separation of epidermis from dermis to evaluate inflammatory mediator production. Following inflammation, rat hind paw epidermis is separated from the dermis by thermolysin at 4 &#176;C. The epidermis is then used for mRNA analysis by RT-PCR and protein evaluation by western blot and immunohistochemistry.

将鼠皮皮和皮米与热解辛分离，以检测部位特异性炎症mRNA和蛋白质

Easy-to-use and inexpensive techniques are needed to determine the site-specific production of inflammatory mediators and neurotrophins during skin ...

免疫与感染

Cheek Injection Model for Simultaneous Measurement of Pain and Itch-related Behaviors

Itch was defined as &#34;an unpleasant cutaneous sensation that provokes a desire to scratch&#34;&#160;by Rothman in 1941. In mouse models, scratch bouts are typically counted to evaluate itch induced by pruritogens. However, previous reports have shown that algesic substances also induce scratching behaviors in a mouse neck injection model, which is the most common test used for scratching behaviors. This finding makes it difficult to study itch in mice. &#160;In contrast, capsaicin, a common algogen, reduced scratching behaviors in some neck injection experiments. Therefore, the effect of pain on scratching behaviors remains unclear. It is thus necessary to develop a method to concurrently investigate itch and pain sensation using behavioral tests. Here, a cheek injection model is introduced which can be used to simultaneously measure pain- and itch-related behaviors. In this model, pruritogens induce scratching behaviors while algesic substances induce wiping behaviors. Using this model, lysophosphatidic acid (LPA), an itch mediator found in cholestatic patients with itch, is shown to exclusively induce itch but not pain. However, in mouse models, LPA has been reported to be both a pruritogen and an algogen. Investigation into the effects of LPA in a mouse cheek injection model showed that LPA only induced scratching, but not wiping behaviors. This indicates that LPA acts as a pruritogen similarly in mice and humans, and demonstrates the utility of a cheek injection model for itch research.

Typically, the mouse neck injection model is used for evaluate pruritogen-induced scratch behaviors. However, the model provides information only on itch, not pain. Here, a cheek injection model is introduced in mice which can be used to simultaneously measure pain and itch-related behaviors.

用于同时测量疼痛和瘙痒相关行为的切面注射模型

Itch was defined as &#34;an unpleasant cutaneous sensation that provokes a desire to scratch&#34;&#160;by Rothman in 1941. In mouse models, scratch bouts ...

神经科学

Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis

Contact hypersensitivity (CHS) is an experimental model of allergic contact dermatitis (ACD) that can be studied in mice. This study aims to present an objective laboratory method that may help to study the CHS reaction in mice, which can be measured and quantified by various tests. To induce CHS, on day "0", mice were sensitized on a previously shaved spot by abdominal skin painting with the hapten 2,4,6-trinitrochlorobenzene (TNCB) in an acetone-ethanol mixture, whereas negative control mice were sham sensitized with vehicle alone-acetone-ethanol mixture. On day "4", the baseline ear thickness was measured with a micrometer prior to the elicitation of CHS (challenge) by painting both ears with diluted TNCB both in the test and control groups. After 24 h, the ear swelling was measured with a micrometer. CHS is an example of a T cell-mediated immune response that causes swelling in inflamed tissue, peaking 24 h after the skin challenge with the same hapten. An increase in ear edema correlated with augmented ear weight, myeloperoxidase (MPO) activity, pro-inflammatory cytokine concentration in the ear extracts, increased thickening of the edematous dermis in the histological examination, and ear vascular permeability. There was also an increase in the concentration of TNP-specific IgG1 antibodies in the sera of the test group when compared with the control mice. Additionally, CHS can be successfully transferred with the CHS-effector cells obtained from donors previously sensitized with TNCB. The CHS-effector cells were administered intravenously into na&#239;ve recipient mice, which were subsequently challenged with the same diluted hapten. Ear swelling was measured with a micrometer 24 h later.

Contact hypersensitivity (CHS) is a murine experimental model of allergic contact dermatitis (ACD). CHS is based on sensitization with reactive hapten by painting the shaved skin of the chest and abdomen, with a subsequent ear skin challenge with a diluted hapten, causing a swelling reaction that is assessed in various ways.

接触超敏反应作为过敏性接触性皮炎的小鼠模型

Contact hypersensitivity (CHS) is an experimental model of allergic contact dermatitis (ACD) that can be studied in mice. This study aims to present an ...

A Mouse Ear Model for Allergic Contact Dermatitis Evaluation

Skin is the human body&#39;s first line of defense and one of the most exposed organs to environmental chemicals. Allergic contact dermatitis (ACD) is a common skin disease that manifests as a local rash, redness, and skin lesions. The occurrence and development of ACD are influenced by both genetic and environmental factors. Although many scholars have constructed a series of models of ACD in recent years, the experimental protocols of these models are all different, which makes it difficult for readers to establish them well. Therefore, a stable and efficient animal model is of great significance to further study the pathogenesis of atopic dermatitis. In this study, we detail a modeling method using 1-fluoro-2,4-dinitrobenzene (DNFB) to induce ACD-like symptoms in the ears of mice and describe several methods for assessing the severity of dermatitis during modeling. This experimental protocol has been successfully applied in some experiments and has a certain promotional role in the field of ACD research.

Here, we describe the methods for inducing allergic contact dermatitis in mouse ears by 1-fluoro-2,4-dinitrobenzene (DNFB) and how to evaluate the severity of allergic contact dermatitis.

用于过敏性接触性皮炎评估的小鼠耳模型

Skin is the human body&#39;s first line of defense and one of the most exposed organs to environmental chemicals. Allergic contact dermatitis (ACD) is a ...

Ex Vivo Release of Calcitonin Gene-Related Peptide from the Trigeminovascular System in Rodents

Calcitonin gene-related peptide (CGRP) was first discovered in the 1980s as a splice variant from the calcitonin gene. Since its discovery, its role in migraine pathophysiology has been well established, first by its potent vasodilator properties and subsequently by its presence and function as a neurotransmitter in the sensory trigeminovascular system. The migraine-provoking ability of CGRP gave support to the pharma industry to develop monoclonal antibodies and antagonists inhibiting the effect of CGRP. A new treatment paradigm has proven effective in the prophylactic treatment of migraine. One of the useful tools to further understand migraine mechanisms is the ex vivo model of CGRP release from the trigeminovascular system. It is a relatively simple method that can be used with various pharmacological tools to achieve know-how to further develop new effective migraine treatments. The present protocol describes a CGRP release model and the technique to quantify the effect of pharmacological agents on the amount of CGRP released from the trigeminovascular system in rodents. A procedure describing the experimental approach from euthanasia to the measurement of protein levels is provided. The essential isolation of the trigeminal ganglion and the trigeminal nucleus caudalis from both mice and rats and the preparation of rat dura mater are described in detail. Furthermore, representative results from both species (rats and mice) are presented. The technique is a key tool to investigate the molecular mechanisms involved in migraine pathophysiology by using various pharmacological compounds and genetically modified animals.

Ex Vivo Release of Calcitonin Gene-Related Peptide from the Trigeminovascular System in Rodents

The present protocol describes the ex vivo calcitonin gene-related peptide (CGRP) release model and the strategy to quantify the effect of pharmacological agents on the amount of CGRP released from the trigeminovascular system in rodents.

离体 啮齿动物三叉血管系统中降钙素基因相关肽的释放

Calcitonin gene-related peptide (CGRP) was first discovered in the 1980s as a splice variant from the calcitonin gene. Since its discovery, its role in ...

<meta charset="utf8"></head><body>为早期产后疼痛研究创建早产大鼠模型

A Preterm Rat Model for Pain Studies

This research delves into the consequences of consistent pinprick stimulation on preterm offspring to ascertain its long-term implications for pain sensitivity. The primary objective of this protocol was to investigate the impact of neonatal pinprick stimuli on the pain threshold in the later stages of life using a preterm rat model. By establishing this model, we aim to advance the research on understanding and managing early postnatal pain associated with prematurity. The findings of this study indicate that while the baseline thresholds to mechanical stimuli remained unaffected, there was a notable increase in mechanical hypersensitivity following complete Freund's adjuvant (CFA) injection in adult rats. Interestingly, compared with male rats, female rats demonstrated heightened inflammatory hypersensitivity. Notably, maternal behavior, the weight of the litters, and the growth trajectory of the offspring remained unchanged by the stimulation. The manifestation of altered nociceptive responses in adulthood after neonatal painful stimuli could be indicative of changes in sensory processing and the functioning of glucocorticoid receptors. However, further research is needed to understand the underlying mechanisms involved and to develop interventions for the consequences of prematurity and neonatal pain in adults.

<meta charset="utf8"></head><body>用于疼痛研究的早产大鼠模型

Here, we present a concise protocol for creating a preterm rat model, facilitating research on early postnatal pain management. The method involves performing a cesarean section three days before the expected birth, extracting preterm rat pups via hysterectomy, and integrating them with the biological offspring of a surrogate dam.

用于疼痛研究的早产大鼠模型

This research delves into the consequences of consistent pinprick stimulation on preterm offspring to ascertain its long-term implications for pain ...

过敏性鼻炎大鼠模型中鼻组织的多色免疫荧光

Immunofluorescent Labeling in Nasal Mucosa Tissue Sections of Allergic Rhinitis Rats via Multicolor Immunoassay

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), affecting approximately 10%-20% of the world's population. While immunofluorescence (IF) staining has long been a standard technique for detecting disease-specific protein expression, conventional IF techniques are limited in their ability to detect the expression levels of three or more proteins in the same sample. Consequently, multicolor IF techniques have been developed in recent years, which allow the simultaneous labeling of multiple targets in cells or tissues.
This protocol provides a comprehensive overview of the process for establishing a rat model of AR, obtaining nasal mucosal samples, and the technical procedures for multicolor immunofluorescence. All rats in the AR group exhibited typical symptoms such as sneezing, a runny nose, and an itchy nose, with behavioral observations scoring &#8805;5 points. Hematoxylin and eosin (H&amp;E) staining revealed increased inflammatory cell counts and disrupted nasal mucosal integrity in the AR group. Multicolor immunofluorescence (mIF) demonstrated increased expression of ROR&#947;t and TICAM-1, while Foxp3 expression decreased in the nasal mucosa tissue of AR rats.

作者聚焦:利用多色免疫荧光推进过敏性鼻炎研究

This protocol describes a multicolor immunofluorescence technique for evaluating the rat model of allergic rhinitis.

通过多色免疫测定法对过敏性鼻炎大鼠鼻粘膜组织切片进行免疫荧光标记

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), ...

甲醛暴露对大鼠新生辣椒素致过敏性皮炎模型的影响

摘要

探索更多视频

此视频中的章节

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

将鼠皮皮和皮米与热解辛分离，以检测部位特异性炎症mRNA和蛋白质

用于同时测量疼痛和瘙痒相关行为的切面注射模型

接触超敏反应作为过敏性接触性皮炎的小鼠模型

用于过敏性接触性皮炎评估的小鼠耳模型

离体啮齿动物三叉血管系统中降钙素基因相关肽的释放

用于疼痛研究的早产大鼠模型

用于疼痛研究的早产大鼠模型

用于疼痛研究的早产大鼠模型

通过多色免疫测定法对过敏性鼻炎大鼠鼻粘膜组织切片进行免疫荧光标记

甲醛暴露对大鼠新生辣椒素致过敏性皮炎模型的影响

摘要

探索更多视频

此视频中的章节

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

将鼠皮皮和皮米与热解辛分离，以检测部位特异性炎症mRNA和蛋白质

用于同时测量疼痛和瘙痒相关行为的切面注射模型

接触超敏反应作为过敏性接触性皮炎的小鼠模型

用于过敏性接触性皮炎评估的小鼠耳模型

离体 啮齿动物三叉血管系统中降钙素基因相关肽的释放

用于疼痛研究的早产大鼠模型

用于疼痛研究的早产大鼠模型

用于疼痛研究的早产大鼠模型

通过多色免疫测定法对过敏性鼻炎大鼠鼻粘膜组织切片进行免疫荧光标记

离体啮齿动物三叉血管系统中降钙素基因相关肽的释放