这项工作是由美国国立卫生研究院授予（HL 098256），由国家和指导研究科学发展奖（K01 AR052840）和独立科学家奖（K02 HL105799）由美国国立卫生研究院授予太平绅士Konhilas和跨学科培训格兰特在心血管科学（HL007249支持）。支持从萨弗心脏中心在亚利桑那大学，距离史蒂芬M. Gootter基金会收到。

所有的实验中，住房和爱护动物中使用该坚持的准则和批准的机构动物护理和使用委员会在亚利桑那大学，并为护理实验动物和使用2011年美国国立卫生研究院准则的协议执行。 1。治疗与4 - 乙烯基环己烯二环氧化物<ol><li> VCD给药方案：在通风橱中，添加0.587毫升VCD到一个干净的瓶中，使终体积10毫升香油，再经反相轻轻地，盖组合和用封口膜密封。将该溶液转移到一个剂量小瓶中，用Parafilm密封，并在4℃下储存长达7天。确保所有试剂均为无菌。 <ol><li>用麻油作为车辆控制，VCD溶于香油。存储VCD（MW =140.2克/摩尔，密度= 0.00克/毫升）在-20℃下</li><li> VCD注射方案：在第2个月的年龄，体重和管理室脾气每日腹腔注射ATURE VCD在160毫克/公斤，连续20天的剂量。在与车辆的溶液，香油类似的方式注射的对照小鼠，在2.5毫升/千克的量。 注：在注射期间，尽量交替注射的部位，以减少局部感染的机会。使用下面的公式来计算所需要的注射量：体重（克）×公斤/ 1,000 GX2.5毫升/公斤=体积（ml）溶液，注入导致大约50毫升/注射用27号针头。由于针的尺寸并不需要局部麻醉剂。 </li><li>发情周期监测：注射周期结束后，通过获得阴道涂片日常监测动情周期。使用背颈背方法抑制小鼠。插入吸管用0.2ml无菌生理盐水到阴道口，以0.5厘米的深度。磨碎盐水（3次），然后将1滴在空白幻灯片。立即评估显微镜使用标准光源microsc样本氧化聚乙烯在100X幅度。 注：阴道脱落细胞学检查是决心确保自行车的损失，因此验证卵巢早衰图2A说明在整个发情周期的不同阶段，典型的阴道细胞学检查的小鼠，即发情前期，发情期，发情后期，间情期17-19。在动情前期，细胞包括有核上皮细胞（ 图2A中，箭头表示的有核细胞）;发情期期间，将细胞主要角化鳞状上皮细胞;动情后期的特征是细胞类型的混合，与白细胞占优势;动情间期是由白细胞为主。焦平面的调整可能需要在整个样本可视化细胞和细胞核。 如图2B所示 ，一个典型的鼠发情周期为4-5天。小鼠经过连续15天的持续性动情考虑非循环。连续地遍及每对研究期间监测动情周期IOD确认缺乏VCD治疗小鼠动情周期。 </li></ol></li></ol> 2。自愿笼轮练习 近交系C57BL / 6和B6C3F1雌性小鼠，年龄7个月，则暴露在自愿笼轮运行。房子个别动物在笼中（47厘米×26厘米×14.5厘米）与不受阻碍的访问到笼子轮28天。 <ol><li>设备设置：笼轮装置包括与5.0厘米宽的行驶面配备了数字磁计数器，该计数器是由轮子转动激活11.5厘米直径的车轮。各车轮行驶试验之前，根据制造商的指导校准笼轮。测量跑轮的内径，计算出圆周，并输入该数值到数字计数器的轮毂尺寸。 <ol><li>附加的数字磁记录计数器的计算机部向轮支架利用胶带。 </li><li>将计算机连接端口离子数字磁计数器车轮支架使用磁带。 </li><li>附加的数字磁记录计数器的自由磁体在相同侧上的中心杆的磁铁通过电缆连接到计算机上，再绑扎充分保护磁铁从咀嚼损坏。磁体的位置应使车轮每旋转记录由计算机没有任何干扰或阻力来完成车轮的转动。 注意：胶带已被成功地用于数字磁计数器的部件附加到传动轮。绑扎应足以完全覆盖磁铁，并防止损坏从由动物咀嚼。磁铁的位置应尽可能接近到车轮而不会妨碍其运动。然而，该设置也应该每天咀嚼伤害监测和修复是必要的。 </li><li>通过滑动轮站通过动物笼子的顶部布线安装车轮的笼子。固定轮架到t用长尾夹他笼接线。 注意：该夹应该是在布线的顶部，远离动物。通过保持器的布线喂计算机部和任何额外的电缆。将电脑和多余电缆动物接触不到的地方。这可通过将一个中等大小的塑料称量在轿厢的布线的顶盘以保持计算机和多余的电缆来实现。 </li></ol></li><li>练习：对于给定的垃圾，随机将小鼠无论是久坐控制或运动养生。给所有的动物和水标准硬盘啮齿动物饲料自由采食。 <ol><li>手动的距离跑（公里/天）和用于运行总时间（小时/天），由磁计数器给定的记录每天的值。记录距离和时间，每天在整个演习期间28天时间准确和一致的价值观相同的1个小时的时间框架内的值。清除（零）磁计数器后，每一天的记录，以确保准确的读数后续ING的日子。 </li><li>计算距离和速度的每个组的平均值。计算速度每天根据距离和时间的给定的值对每个小鼠。数据可以为每个测量参数作为每天或每周的平均值显示。 </li></ol></li><li>组织收获：在具体行使期结束时，安乐死内下吸入麻醉去除鸡笼颈椎脱位的30分钟行使和久坐对照组动物。称量体重，并迅速消费心，骨骼肌和卵巢。 <ol><li>在组织收获的时候，装载和弄脏收集卵巢与苏木精和伊红（H &amp;E; 图2C）。在放大镜下，计数和每个VCD治疗和控制动物20归类为毛囊原始，初级，中级，或胃窦部。甲原始卵泡包含一个小的初级卵母细胞，扁平或鳞状颗粒细胞密切相对于卵母细胞的单个层，以及基底椎板。一个主要的卵泡是由一个或多个立方形颗粒细胞的存在下被布置在围绕卵母细胞的单层限定。次级卵泡包含周围形成多个卵母细胞层和卵泡膜颗粒细胞。一个窦状卵泡的特征在于，含有流体的空腔或“腔”。 </li><li>切除的心脏，使内侧切口的腹侧和打开隔膜后可视化的心脏。在该基地并将其放置在冷磷酸盐缓冲液充分洗出残留心室的血液迅速切除心脏。修剪多余的组织的心脏，权衡整个心脏和捕捉冻结在液氮中。 </li><li>用改性冰冷的磷酸盐缓冲盐水溶液（PBS）洗涤所有其他切除的组织，和扣冻结在异戊烷在液氮中冷却。 </li></ol></li></ol> 3。强迫跑步机练习<ol><li>当采用强迫运动范式吨Ø测量运动能力，使用一台跑步机，并设置如下： <ol><li>将电击电网在跑步机上斜坡的底部。产生这种冲击作为一个刺激的运动。的电刺激的强度和频率是手动的使用者控制。网格可以启用或禁用单独为每个车道。所有数据收集是手动完成的。 </li><li>周期性地，根据制造商的指导进行校准的跑步机。 <ol><li>确保在跑步机控制器前面板切换设置为“STOP”。 </li><li>上跑步机带泳道1地点转速传感器和安全地与螺母拧紧在如下图所示的位置。 </li><li>使用拨号的跑步机控制面板上，调整速度887米/分钟之后，跑步机控制器面板上设置的电刺激频率为2 Hz的频率。 </li><li>按“里程表复位”按钮来初始化校准程序。 <strong&gt;的注意：久坐或nonexercise对照组必须包含在实验设计。久坐组为年龄匹配的运动组。之前我们已经表明，个别房屋的小鼠久坐在同样大小的笼子到行使小鼠相比，安置与兄弟姐妹久坐小鼠没有影响所需的参数。因此，久坐老鼠被安放在同一个房间内行使小鼠的兄弟姐妹。 </li></ol></li></ol></li><li>驯化：之前每天的实验3-7天驯化老鼠的跑步机。要做到这一点，地方的小鼠与带不动的震撼和电网断开，但对（噪声），带电机的跑步机。离开静置至少15分钟的小鼠。 </li><li>热身：热身如本协议中使用如下的步骤： <ol><li>将每只小鼠在跑步机上。禁用电网的冲击，提高带速4米/分钟持续15分钟。 </li><li>使电网的冲击，保持皮带速度4米/分钟的皮带速度为10分钟。 </li></ol></li><li>运动训练方案<ol><li>以下热身，不断冲击电网和增加皮带速度5米/分钟持续15分钟。 </li><li>提高带速为15米/分钟额外15分钟。 </li><li>下面的训练，转移小鼠回到自己的笼子。 注：在本研究中，运动训练1周每日执行。 </li><li>急性运动疗法：急性运动养生的目的是确定极端运动负荷的短期和长期影响。 <ol><li>以下预热，增加皮带速度为5米/分钟到10，15，20，25，和30米/分钟在10分钟的间隔。 注：终止运动，如果老鼠是无法继续疲惫和痛苦的运行或迹象。在我们的研究中，大多数动物都停在25米/分钟，无动物通过了30米/分的阶段。 </li><li>测试每个鼠标三次，让每个测试之间的2-3天。所有埃克塞计算平均值精确的会话每只小鼠。 </li></ol></li><li>运动养生耐力：耐力运动协议旨在培养老鼠的长期暴露在跑步机上运动的刺激。这种类型的协议测试容忍适度的运动应力的长期影响的能力。 <ol><li>首先在热身期间，4米/分钟概述的皮带速度。 </li><li>逐步提高皮带速度1m/min卷材，直到20米的最终速度/分钟为止。一个典型的研究方案的总时间为50分钟。 </li><li>运动养生后，小鼠转回到自己的笼子。 注意：运行小鼠消耗体力是可以避免的。如果在实验过程中的任何一点鼠标显示疲惫的迹象，停用电网的冲击，让鼠标休息和恢复。 </li></ol></li></ol></li></ol> 4。数据和统计分析<ol><li>本研究结果为平均值±标准差，并使用双因子ANOVA（治疗组和exercise为主要因素），其次是学生，纽曼Keuls检验事后检验或学生的t检验来比较平均值之间的差异。 </li><li>由每个行使动物的质量进行比较，以久坐组的平均心脏质量测定心脏重量与运动变化百分比。心脏质量差被表示为久坐不动的动物的百分比变化为每个相应的动物。的&lt;0.05的p值被认为是统计学显著。 </li></ol>

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	<li>Lindheim, S. R., 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=Comparison+of+estimates+of+insulin+sensitivity+in+pre-+and+postmenopausal+women+using+the+insulin+tolerance+test+and+the+frequently+sampled+intravenous+glucose+tolerance+test.">Comparison of estimates of insulin sensitivity in pre- and postmenopausal women using the insulin tolerance test and the frequently sampled intravenous glucose tolerance test.</a> J. Soc. Gynecol. Invest. 1, 150-154 (1994).</li><li>Dubnov-Raz, G., Pines, A., Berry, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diet+and+lifestyle+in+managing+postmenopausal+obesity.">Diet and lifestyle in managing postmenopausal obesity.</a> Climacteric. 10 Suppl 2, 38-41 (2007).</li><li>Grundy, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+syndrome:+connecting+and+reconciling+cardiovascular+and+diabetes+worlds.">Metabolic syndrome: connecting and reconciling cardiovascular and diabetes worlds.</a> J. Am. Coll. Cardiol. 47, 1093-1100 (2006).</li><li>Sharma, S., Firoozi, S., McKenna, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Value+of+exercise+testing+in+assessing+clinical+state+and+prognosis+in+hypertrophic+cardiomyopathy.">Value of exercise testing in assessing clinical state and prognosis in hypertrophic cardiomyopathy.</a> Cardiol. Rev. 9, 70-76 (2001).</li><li>Guyatt, G. H., Devereaux, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+heart+failure+treatment.">A review of heart failure treatment.</a> Mt. Sinai. J. Med. 71, 47-54 (2004).</li><li>Mayer, L. P., 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=Long-term+effects+of+ovarian+follicular+depletion+in+rats+by+4-vinylcyclohexene+diepoxide.">Long-term effects of ovarian follicular depletion in rats by 4-vinylcyclohexene diepoxide.</a> Reprod. Toxicol. 16, 775-781 (2002).</li><li>Mayer, L. P., Devine, P. J., Dyer, C. A., Hoyer, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+follicle-deplete+mouse+ovary+produces+androgen.">The follicle-deplete mouse ovary produces androgen.</a> Biol. Reprod. 71, 130-138 (2004).</li><li>Lohff, J. C., Christian, P. J., Marion, S. L., Hoyer, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+duration+of+dosing+on+onset+of+ovarian+failure+in+a+chemical-induced+mouse+model+of+perimenopause.">Effect of duration of dosing on onset of ovarian failure in a chemical-induced mouse model of perimenopause.</a> Menopause. 13, 482-488 (2006).</li><li>Wright, L. 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=Comparison+of+skeletal+effects+of+ovariectomy+versus+chemically+induced+ovarian+failure+in+mice.">Comparison of skeletal effects of ovariectomy versus chemically induced ovarian failure in mice.</a> J. Bone Miner Res. 23, 1296-1303 (2008).</li><li>Ogawa, S., Chan, J., Gustafsson, J. A., Korach, K. S., Pfaff, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrogen+increases+locomotor+activity+in+mice+through+estrogen+receptor+alpha:+specificity+for+the+type+of+activity.">Estrogen increases locomotor activity in mice through estrogen receptor alpha: specificity for the type of activity.</a> Endocrinology. 144, 230-239 (2003).</li><li>Roy, E. J., Wade, G. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+estrogens+in+androgen-induced+spontaneous+activity+in+male+rats.">Role of estrogens in androgen-induced spontaneous activity in male rats.</a> J. Comp. Physiol. Psychol. 89, 573-579 (1975).</li><li>Fahrbach, S. E., Meisel, R. L., Pfaff, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preoptic+implants+of+estradiol+increase+wheel+running+but+not+the+open+field+activity+of+female+rats.">Preoptic implants of estradiol increase wheel running but not the open field activity of female rats.</a> Physiol. Behav. 35, 985-992 (1985).</li><li>Morgan, M. A., Pfaff, D. W. <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+estrogen+on+activity+and+fear-related+behaviors+in+mice.">Effects of estrogen on activity and fear-related behaviors in mice.</a> Horm. Behav. 40, 472-482 (2001).</li><li>Gorzek, J. F., 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=Estradiol+and+tamoxifen+reverse+ovariectomy-induced+physical+inactivity+in+mice.">Estradiol and tamoxifen reverse ovariectomy-induced physical inactivity in mice.</a> Med. Sci. Sports Exerc. 39, 248-256 (2007).</li><li>Kadi, F., 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+effects+of+physical+activity+and+estrogen+treatment+on+rat+fast+and+slow+skeletal+muscles+following+ovariectomy.">The effects of physical activity and estrogen treatment on rat fast and slow skeletal muscles following ovariectomy.</a> J. Muscle Res. Cell Motil. 23, 335-339 (2002).</li><li>Konhilas, J. P., 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=Sex+modifies+exercise+and+cardiac+adaptation+in+mice.">Sex modifies exercise and cardiac adaptation in mice.</a> Am. J. Physiol. Heart Circ. Physiol. 287, 2768-2776 (2004).</li><li>Caligioni, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+reproductive+status/stages+in+mice.">Assessing reproductive status/stages in mice.</a> Curr. Protoc. Neurosci. 4, (2009).</li><li>Goldman, J. M., Murr, A. S., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rodent+estrous+cycle:+characterization+of+vaginal+cytology+and+its+utility+in+toxicological+studies.">The rodent estrous cycle: characterization of vaginal cytology and its utility in toxicological studies.</a> Birth Defects Res. B. Reprod. Toxicol. 80, 84-97 (2007).</li><li>McLean, A. C., Valenzuela, N., Fai, S., Bennett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performing+vaginal+lavage,+crystal+violet+staining,+and+vaginal+cytological+evaluation+for+mouse+estrous+cycle+staging+identification.">Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification.</a> J. Vis. Exp. , (2012).</li><li>McKee, L. 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=Sexually+dimorphic+myofilament+function+and+cardiac+troponin+I+phosphospecies+distribution+in+hypertrophic+cardiomyopathy+mice.">Sexually dimorphic myofilament function and cardiac troponin I phosphospecies distribution in hypertrophic cardiomyopathy mice.</a> Arch. Biochem. Biophys. 535, 39-48 (2013).</li><li>Konhilas, J. P., 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=Exercise+can+prevent+and+reverse+the+severity+of+hypertrophic+cardiomyopathy.">Exercise can prevent and reverse the severity of hypertrophic cardiomyopathy.</a> Circ. Res. 98, 540-548 (2006).</li><li>Perez, N. J., Chen, H., Regan, J. A., Emert, A., Constantopoulos, E., Lynn, M., Konhilas, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+chemically-induced+ovarian+failure+on+voluntary+cage+wheel+exercise+and+cardiac+adaptation+in+mice.">The impact of chemically-induced ovarian failure on voluntary cage wheel exercise and cardiac adaptation in mice.</a> Compar. Med. In press, (2013).</li></ol>

作者什么都没有透露。

两个笼子轮运动和跑步机上跑步运动疗法已被用于在我们以前的研究16,21,22。所有的动物都保存在专门的运动室，以尽量减少对环境或人类的干扰。动物的任何干扰“生物节律可以显著影响动物的运动性能。因此，整个动物保健，性能监控和记录，以及运动疗法应在每天相同的时间框架内完成。调查人员应尽量避免额外的压力对动物通过展示的预防措施，如限制应激暴露的持续时间和覆盖的笼子在适当的时候。 凯奇转轮运动 当准备笼子轮装置，车轮应该是方便的鼠标，安装并退出。应在笼的底部最小的床上用品，以防止老鼠从嵌套围绕轮盘和阻碍频率Ë轮运动。磁传感器被放置在轮的固定部分和固定到所述自由轮的磁体，以便计数轮每转一圈的鼠标行使。附着在磁传感器记录装置应放置在上述保持器的电线盖，及所有裸露的电线必须要么胶带或保持够不着从小鼠中，以便提供对咀嚼损伤的屏障。磁体和磁传感器也被贴在适当的结构以防止二者损坏和不希望的运动。在日常的数据收集，所有的录音线必须密切检查，以确保电线保持完好，并应retaped是必要的。因为它可能需要数天的小鼠，以适应新的环境，并开始定期锻炼，数据记录应针对每个个体动物。虽然动物往往要花费许多人力资源上的车轮，大部分的运行发生在夜间。因此，一些动物可能揭示HIGH性能的活动，即使他们出现在白天蛰伏。此外，它的情况并不少见，有些动物会表现出有限的笼子轮运行。通常情况下，笼轮的活动，相当于平均1公里/天以下是随机的笼子里活动的结果，并没有与任何显著运行的刺激有关。因此，动物的运行参数为1千米/天或1小时/天或更少应该被排除在最终的数据分析nonrunners。 取决于个体的研究，涉及小鼠的菌株的具体目的，运动的长度可以改变。我们以前的研究表明，对于涉及两个小鼠品系，平均轮运行速度和运动上笼轮的持续时间逐渐增加随着时间的推移和4周趋于​​稳定。 凯奇轮运转是在动物模型中一种自愿性的锻炼，这是一般取为压力较小的情况下被执行。 UNL其中某些参数可预置IKE跑步机上锻炼，在测试笼子里所有正在运行的是自愿的，容易受环境或行为线索。因此，通常可以避免不同的实验设置之间的比较，除非使用大型实验组号码。其他潜在的研究领域，可以从这种类型的活动范式的受益是把行为反应与生理后果，如认知神​​经科学的研究。对于利用自愿笼轮子范式的研究中，最关键的步骤是笼设置和监测工作的活动。 跑步机上锻炼 驯化步骤是必须的准备工作动物在跑步机上锻炼的协议。取决于个体研究的具体目的，驯化期间的持续时间可以变化。在这项研究中，为期一周的被选择让动物acclim吃了跑步机设备和电机的噪声在笼子里，热身期间是运动训练方案的一个重要步骤。类似人类同行，老鼠必须进入任何剧烈运动疗法，以避免受伤和潜在的数据工件前热身。在这项研究中，我们开始带以尽可能低的速度（4米/分钟）与电网的冲击关闭15分钟。所涉及的所有小鼠开始运行和关闭的皮带。震荡网格然后接通和皮带速度​​保持在4米/分钟。为了避免过度强调小鼠，我们设置了电击在温和的水平（1赫兹）。在几天内，所有小鼠适应了跑步机皮带。 行驶的持续时间以及加速度，也可以针对每个实验设计。推荐的启动加速度为1米/分钟。小鼠呈现枯竭的迹象的跑步机通道，必须立即关闭。疲惫的迹象通常由英里展出行政长官在跑步机上载列如下： <ol><li>连续暴露在冲击电网&gt; 5-10秒， </li><li>运行能力造成冲击电网曝光&gt; 5倍单运动养生期间和损失</li><li>运行能力造成冲击电网暴露的损失单一的运动处方中&gt; 2秒的至少3倍。 </li></ol>上面列出的值将被用于主要是作为一个特定的实验指导方针。这些参数需要为每一个练习的协议确定，因为它从不同小鼠品系小鼠品系，实验进行实验，并调查调查。 不像笼轮运动，跑步机的研究必须重复几次，以消除对环境和生物因素上运行的性能的影响。这是并不少见的，即使是同一种动物的测试运行之间的性能波动。由于每个运动养生自曝运行小鼠的STREβ，测试之间的适当的休息间隔是必要的。根据我们的经验，2-3天每次测试之间的休息时间为足以让老鼠恢复。 如同人类的跑步机上锻炼时，速度耐力运动的心脏率或血氧饱和度，一个技术上的困难的壮举与小鼠的​​研究是最好的决定。跑步研究中最关键的步骤是判定用尽，这通常单个小鼠之间变化的，并且被设置为80％的最大速度。在我们自己的经验结果的记录可能个别研究人员之间的显著差异，因为标准的疲惫大多是主观的。因此，相同的标准在记录过程中一致的应用是至关重要的。

自然发病更年期的特点是卵巢原始卵泡通过闭锁的枯竭，最终导致卵巢衰老。在美国，一个妇女一生中超过30％将用于绝经后。为应对卵巢功能下降，一些生理和心理后果可能发生，包括增加肥胖，血管不稳定，情绪或睡眠障碍。因为肥胖/肥胖的增加而绝经前对口1,2相比，绝经后的妇女特别易受影响代谢综合征和相关的并发症，包括心血管疾病（CVD）3。此外，它正变得越来越明显的是，成功的心脏康复患者的CVD包括规律的有氧运动，而且运动可以降低心血管疾病发病率和死亡率在这些科目4,5。然而，如何运动能力茶在售前到售后，绝经过渡期民营企业还没有得到很好的研究。更重要的是，运动作为一种预防性措施，在绝经后妇女心血管疾病风险的作用仍然相当充分研究。 用于妇女绝经模仿最常用的诱导模型是卵巢切除（OVX），啮齿目动物。近日，职业性化学性4 -乙烯基环己烯二环氧化物（VCD）已被证明通过加快闭锁的自然过程，最终导致与在其他组织中6未发现明显毒性卵巢功能衰竭专门针对小规模小学和原始卵泡。 VCD治疗的小鼠的动情周期，这是类似于一个人的月经周期，完成VCD注射停止后2-3个月内，这反过来又导致雌激素的逐渐撤离，模仿围与绝经后过渡。因此，卵泡消耗臭氧层，卵巢完好的动物十分接近人类自然通过发展预以近绝经后过渡7-9。此外，它提供了一种非手术的替代OVX模型减少手术的并发症，如感染。在这项研究中，我们采用VCD诱发更年期小鼠来研究运动对更年期小鼠心脏适应的影响。 有迹象表明，建议在男女双方的鼠类10-13雌激素和保持架转轮运动之间的关联几个实例。雌激素耗竭手术去势降低小鼠和大鼠14,15自愿演习活动。练习用两种方法在这项研究中，测试VCD诱发更年期小鼠的运动能力。凯奇轮运转被普遍认为是在动物模型中一种自愿性的锻炼和表现，想必，压力较小的情况下。然而，笼子轮运行并不表示动物的相对运动能力，这就要求对动物的健身活动SE在更高的层次。强迫跑步锻炼是用来确定运动能力，衡量一个宽容运动强度和耐力。此外，我们先前已经表明，笼转轮运动提供了一个刺激心肌肥厚和增生性这是成长性依赖16。所以，我们还测量了心脏适应于VCD诱发更年期笼的小鼠转轮运动。

<table border="0" cellpadding="0" cellspacing="0" width="682">
	<colgroup>
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr>
			<td>4-vinylcyclohexene diepoxide (VCD)</td>
			<td>Sigma, cat# V-3630</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>11.5-cm-diameter wheel with a 5.0-cm-wide running surface</td>
			<td>Petsmart,model 6208</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Digital magnetic counter</td>
			<td>Sigma Sport,model BC 600</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Treadmill exercise</td>
			<td>Columbus Instruments , model 1055SDRM</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a method to study the impact of chemically-induced ovarian failure on exercise capacity and cardiac adaptation in mice

心血管疾病（CVD）的风险增加了绝经后的妇女，但是，锻炼的作用，作为一种预防性措施，在绝经后妇女心血管疾病的风险并没有得到充分研究。因此，我们研究了自愿笼轮运动和心脏适应更年期小鼠强迫跑步机上锻炼的影响。用于妇女绝经模仿最常用的诱导模型是卵巢切除（OVX），啮齿目动物。然而，去势模型从更年期在人类几相异性。在这项研究中，我们给予4 - 乙烯基环己烯二环氧化物（VCD）到雌性小鼠，从而加速卵巢功能衰竭的替代更年期模型来研究在绝经期小鼠运动的影响。 VCD选择性地加速了初级和原始卵泡导致的内分泌状态密切从模仿自然进程的损失预以近至绝经后的人类。要确定运动对运动的影响能力和心脏适应VCD治疗的雌性小鼠，两种方法被使用。首先，我们曝光了一组视像处理和未经处理的小鼠对自愿笼轮。第二，我们被迫使用跑步机运动，以确定测量公差运动强度和耐力独立成团VCD处理和未经处理的小鼠运动能力。

一个典型的实验方案，如在本研究中使用的，示于图1。继VCD治疗连续20天，阴道细胞学检查是用来确定在VCD注入小鼠体内是否存在周期性的;媒介物注射的小鼠也进行监测。在发情周期的阶段是由上皮细胞，角化上皮细胞，白细胞和在阴道涂片（ 图2A）的比例确定。仅角质化上皮细胞的存在所指示的鼠标是在发情期，因此仍然骑自行车。动情周期小鼠一般持续4天，因此VCD注射的动物被认为是后无环执着动情，15天中，阴道细胞学检查发现白细胞的大部分（ 图2A）。 VCD治疗的小鼠的动情周期变得不规则，再VCD注射开始（ 图2B）后2-3个月内停止</STRONG&gt;。 VCD卵巢组织呈显著萎缩，与对照组相比。在高倍镜下，我们看到小学和原始卵泡（ 图2C）的完全耗尽。 为了测试VCD诱发卵巢早衰的运动表现的影响，我们随机7个月旧VCD和媒介物处理的小鼠（双株，C57BL / 6和B6C3F1）要么久坐或笼轮运动组4-6周后停止循环（ 图1）。整个4周的运动期间，记录每天的锻炼价值的时间和距离对于每个行使动物。日常运行中值的距离，时间和速度的每周的平均值示于图3为与车辆或VCD处理的C57BL / 6小鼠。我们没有看到由平均每天的时间，距离和速度上在任C57BL / 6或B6C3F1鼠标VCD和车辆处理组之间的笼测量车轮运动表现显著差异菌株（ 图4A）。计算出的车轮运行的速度超过先前16所示的4个星期的运行周期逐渐增多，但并没有实验组之间的显著差异。这些研究表明VCD引起的卵巢功能衰竭自愿笼转轮运动的影响超越了至少2个小鼠品系。 继笼自愿转轮运动协议的期限，身体形态测量，记录和心脏被迅速切除，称重。之前我们已经表明，自愿笼转轮运动诱发增加心脏重量16。在这里，我们也显示心肌肥厚响应笼转轮运动是绝对的心脏重量（HW）测量和硬件标准化，以胫骨长度（TL）（ 表1）。相比同行久坐的绝对心脏质量和HW / TL比值均显著更大的车辆和VCD治疗的小鼠。然而，没有measurable差异控制和VCD引起的卵巢功能衰竭的小鼠下笼自愿转轮运动之间的心肌肥厚。 接下来，我们使用非自愿（强制）跑步机上运行，​​以确定运动能力。我们首先进行小鼠对高强度的协议，增加平均速度逐步每10分钟直到筋疲力尽。平均来说，载体处理的小鼠的最大速度为26.7±0.95米/分钟和VCD处理的小鼠的最大速度为用尽前28±1.4米/分钟。我们在使用运行20米/分钟（约80％最大），以测定对续航能力较低强度。 如图4B所示 ，在高或低强度的运动能力无显著差异车辆和视像处理组之间被发现，表明VCD诱导的卵巢功能衰竭，对小鼠的运动能力没有影响。 总之，更年期引起的VCD疗法对无论是自愿或被迫R无影响<html> unning容量，以及在心脏的适应性反应来锻炼。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig1highres.jpg" src="/files/ftp_upload/51083/51083fig1.jpg" /> 图1：VCD-管理和实验方案。小鼠施用VCD（160毫克/千克）或香油（车辆控制）开始于2月龄，连续20天。以下VCD或车辆注射，动情周期监测和旋回分别如在方法部分中描述的确定。卵巢功能丧失发生在60-90天，是由阴道细胞学检查确诊。 7个月的年龄，老鼠被暴露在自愿笼转轮运动4周或遭受跑步机运动方案。 oad/51083/51083fig2highres.jpg“SRC =”/ files/ftp_upload/51083/51083fig2.jpg“/&gt; 。小鼠图2 VCD诱导卵巢功能衰竭答：从车辆注入小鼠表现出发情周期的四个阶段未染色阴道涂片细胞学：1）发情前期，主要包括有核上皮细胞2）发情期，其特点是未-成核的角化细胞; 3）发情后期，包括有核上皮细胞（箭头），未核角化上皮细胞（箭头）和白细胞; 4）动情，白细胞主要包括;和B：车辆和VCD注射组的发情周期平均长度（天）。一个周期是从在发情期的第一天测到在发情期下一期的第一天。阴道细胞学开始大约6周后注射的发作。 VCD-注射的动物的动情周期变得不规律，比注射赋形剂的动物s，而12后的发情周期，所有的VCD细胞学注射的动物不再显示循环C：H＆E在8月龄卵巢组织染色。横截面为5μm的厚度做了车辆处理（左图）和VCD处理（右图）小鼠20​​X（上图）和200X（下图）的放大倍率。在放大200倍的卵巢组织显示了一些在车辆相比，VCD治疗的小鼠卵泡。 <a href="https://www.jove.com/files/ftp_upload/51083/51083fig2highres.jpg" target="_blank">请点击这里查看这个数字的放大版本。</a> <img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig3highres.jpg" src="/files/ftp_upload/51083/51083fig3.jpg" /> 图3：在车辆或VCD的处理C57BL/6J雌性小鼠自愿笼轮的表现。左图：平均跑动距离（公里/天），每1周的时间超过4周的研究期间中间面板：平均运行时间（小时/天），每1周的时间超过4周的研究期间右面板：平均运行速度（公里/小时），每1周时间内超过4周的研究期间， <a href="https://www.jove.com/files/ftp_upload/51083/51083fig3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig4highres.jpg" src="/files/ftp_upload/51083/51083fig4.jpg" /> 图4。自愿笼车轮和车辆与VCD治疗的小鼠跑步机的性能。答：（上图）的C57BL / 6平均行驶距离（公里/日）和时间，每24小时超过4周的研究期间花在轮（小时/天）。 （ 下图）＆＃160; B6C3F1平均行驶距离（公里/日）和时间，每24小时超过4周的研究期间 B花在轮（小时/天）：C57BL / 6跑步机运行参数。最大使用时间的方法部分的详细方案（ 上图）和最大速度（Botoom面板 ）。

Watch this Scientific Journal Video about A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice at JoVE.com

A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

两个运动范式是一个新开发的化学诱导更年期小鼠模型测试，以检查更年期对运动能力和心脏的适应锻炼的影响。

一个方法研究的化学诱导卵巢早衰的运动能力和适应心脏的影响在小鼠

The risk of cardiovascular disease (CVD) increases in post-menopausal women, yet, the role of exercise, as a preventative measure for CVD risk in ...

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Research

JoVE Journal

Medicine

15.1K 次观看.  University of Arizona. 两个运动范式是一个新开发的化学诱导更年期小鼠模型测试，以检查更年期对运动能力和心脏的适应锻炼的影响。 

视频: A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo and noninvasively in humans via phosphorus-31 magnetic resonance spectroscopy (31PMRS). However, the approach has not been widely adopted in translational and clinical research, with variations in methodology and limited guidance from the literature. Increased optimization, standardization, and dissemination of methods for in vivo 31PMRS would facilitate the development of targeted therapies to improve OXPHOS capacity and could ultimately favorably impact cardiovascular health. 31PMRS produces a noninvasive, in vivo measure of OXPHOS capacity in human skeletal muscle, as opposed to alternative measures obtained from explanted and potentially altered mitochondria via muscle biopsy. It relies upon only modest additional instrumentation beyond what is already in place on magnetic resonance scanners available for clinical and translational research at most institutions. In this work, we outline a method to measure in vivo skeletal muscle OXPHOS. The technique is demonstrated using a 1.5 Tesla whole-body MR scanner equipped with the suitable hardware and software for 31PMRS, and we explain a simple and robust protocol for in-magnet resistive exercise to rapidly fatigue the quadriceps muscle. Reproducibility and feasibility are demonstrated in volunteers as well as subjects over a wide range of functional capacities.

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

This work demonstrates the feasibility of an in vivo phosphorus-31 magnetic resonance spectroscopy (31PMRS) technique to quantify mitochondrial oxidative phosphorylation (OXPHOS) capacity in human skeletal muscle.

磷31磁共振波谱分析：一种测量工具<em&gt;在体内</em&gt;线粒体氧化磷酸化能力人骨骼肌

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...

科研

医学

Impact of Intracardiac Neurons on Cardiac Electrophysiology and Arrhythmogenesis in an Ex Vivo Langendorff System

Since its invention in the late 19th century, the Langendorff ex vivo heart perfusion system continues to be a relevant tool for studying a broad spectrum of physiological, biochemical, morphological, and pharmacological parameters in centrally denervated hearts. Here, we describe a setup for the modulation of the intracardiac autonomic nervous system and the assessment of its influence on basic electrophysiology, arrhythmogenesis, and cyclic adenosine monophosphate (cAMP) dynamics. The intracardiac autonomic nervous system is modulated by the mechanical dissection of atrial fat pads-in which murine ganglia are located mainly&#8212;or by the usage of global as well as targeted pharmacological interventions. An octapolar electrophysiological catheter is introduced into the right atrium and the right ventricle, and epicardial-placed multi-electrode arrays (MEA) for high-resolution mapping are used to determine cardiac electrophysiology and arrhythmogenesis. F&#246;rster resonance energy transfer (FRET) imaging is performed for the real-time monitoring of cAMP levels in different cardiac regions. Neuromorphology is studied by means of antibody-based staining of whole hearts using neuronal markers to guide the identification and modulation of specific targets of the intracardiac autonomic nervous system in the performed studies. The ex vivo Langendorff setup allows for a high number of reproducible experiments in a short time. Nevertheless, the partly open nature of the setup (e.g., during MEA measurements) makes constant temperature control difficult and should be kept to a minimum. This described method makes it possible to analyze and modulate the intracardiac autonomic nervous system in decentralized hearts.

Impact of Intracardiac Neurons on Cardiac Electrophysiology and Arrhythmogenesis in an Ex Vivo Langendorff System

Here, we present a protocol for the modulation of the intracardiac autonomic nervous system and the assessment of its influence on basic electrophysiology, arrhythmogenesis, and cAMP dynamics using an ex vivo Langendorff setup.

心内神经元对体外 Langendorff 系统心脏电生理和心脏交感神经的影响

Since its invention in the late 19th century, the Langendorff ex vivo heart perfusion system continues to be a relevant tool for studying a broad spectrum ...

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies that promote efficient bone healing are non-existent and in high demand. Effective and reproducible animal models of fractures healing are needed to understand the complex biological processes associated with bone regeneration. Many animal models of fracture healing have been generated over the years; however, murine fracture models have recently emerged as powerful tools to study bone healing. A variety of open and closed models have been developed, but the closed femoral fracture model stands out as a simple method for generating rapid and reproducible results in a physiologically relevant manner. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice and facilitate a post-fracture stabilization of the femur by inserting an intramedullary steel rod. Although devices such as a nail or a screw offer greater axial and rotational stability, the use of an intramedullary rod provides a sufficient stabilization for consistent healing outcomes without producing new defects in the bone tissue or damaging nearby soft tissue. Radiographic imaging is used to monitor the progression of callus formation, bony union, and subsequent remodeling of the bony callus. Bone healing outcomes are typically associated with the strength of the healed bone and measured with torsional testing. Still, understanding the early cellular and molecular events associated with fracture repair is critical in the study of bone tissue regeneration. The closed femoral fracture model in mice with intramedullary fixation serves as an attractive platform to study bone fracture healing and evaluate therapeutic strategies to accelerate healing.

The murine closed femoral fracture model is a powerful platform to study fracture healing and novel therapeutic strategies to accelerate bone regeneration. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice using an intramedullary steel rod to stabilize the femur.

小鼠闭合性股骨骨折的形成: 骨愈合模型的研究

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

进行最大和次极大耐力运动测试, 测量人类急性运动的生理和生物学反应

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

小鼠肌功能的无创评价

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

Generation and Characterization of Right Ventricular Myocardial Infarction Induced by Permanent Ligation of the Right Coronary Artery in Mice

Right ventricular infarction (RVI) is a common presentation in clinical practice. Severe RVI can lead to fatal hemodynamic dysfunction and arrhythmia. In contrast to the extensively used mouse myocardial infarction (MI) model generated by left coronary artery ligation, the RVI mouse model is rarely employed due to the difficulty associated with model generation. Research on the mechanisms and treatment of RVI-induced RV remodeling and dysfunction requires animal models to mimic the pathophysiology of RVI in patients. This study introduces a feasible procedure for RVI model generation in C57BL/6J mice. Further, this model was characterized based on the following: infarct size evaluation at 24 h after MI, assessment of cardiac remodeling and function with echocardiography, RV hemodynamics assessment, and histology of the infarct zone at 4 weeks after RVI. In addition, a coronary vasculature cast was performed to observe the coronary arterial arrangement in RV. This mouse model of RVI would facilitate the research on mechanisms of right heart failure and seek new therapeutic targets of RV remodeling.

There are several differences between the right and left ventricles. However, the pathophysiology of right ventricular infarction (RVI) has not been clarified. In the present protocol, a reproducible method for RVI mouse model generation is introduced, which may provide a means to explain the mechanism of RVI.

小鼠右冠状动脉永久结扎诱导的右心室性心肌梗死的生成与表征

Right ventricular infarction (RVI) is a common presentation in clinical practice. Severe RVI can lead to fatal hemodynamic dysfunction and arrhythmia. In ...

A Piglet Perinatal Asphyxia Model to Study Cardiac Injury and Hemodynamics after Cardiac Arrest, Resuscitation, and the Return of Spontaneous Circulation

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model by introducing cardiac arrest. This enabled us to study the impact of severe asphyxia on key outcomes, including the time taken for the return of spontaneous circulation (ROSC), as well as the effect of chest compressions according to alternative protocols for cardiopulmonary resuscitation. Due to the anatomical and physiological similarities between piglets and human neonates, piglets serve as good models in studies of cardiopulmonary resuscitation and hemodynamic monitoring. In fact, this cardiac arrest model has provided evidence for guideline development through research on resuscitation protocols, pathophysiology, biomarkers, and novel methods for hemodynamic monitoring. Notably, the incidental finding that a substantial fraction of piglets have pulseless electrical activity (PEA) during cardiac arrest may increase the applicability of the model (i.e., it may be used to study pathophysiology extending beyond the perinatal period). However, the model generation is technically challenging and requires various skill sets, dedicated personnel, and a fine balance of the measures, including the surgical protocols and the use of sedatives/analgesics, to ensure a reasonable rate of survival. In this paper, the protocol is described in detail, as well as experiences with adaptations to the protocol over the years.

This piglet model involves surgical instrumentation, asphyxiation until the cardiac arrest, resuscitation, and post-resuscitation observation. The model allows for multiple sampling per animal, and by using continuous invasive arterial blood pressure, ECG, and non-invasive cardiac output monitoring, it provides knowledge about hemodynamics and cardiac pathophysiology in perinatal asphyxia and neonatal cardiopulmonary resuscitation.

仔猪围产期窒息模型，用于研究心脏骤停、复苏和自主循环恢复后的心脏损伤和血流动力学

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model ...

Chronic Ovine Model of Right Ventricular Failure and Functional Tricuspid Regurgitation

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to suboptimal clinical results. We set out to establish a chronic ovine model of FTR and right heart failure to investigate the mechanisms of FTR. Twenty adult male sheep (6-12 months old, 62 &#177; 7 kg) underwent a left thoracotomy and baseline echocardiography. A pulmonary artery band (PAB) was placed and cinched around the main pulmonary artery (PA) to at least double the systolic pulmonary artery pressure (SPAP), inducing right ventricular (RV) pressure overload and signs of RV dilatation. PAB acutely increased the SPAP from 21 &#177; 2 mmHg to 62 &#177; 2 mmHg. The animals were followed for 8 weeks, symptoms of heart failure were treated with diuretics, and surveillance echocardiography was used to assess for pleural and abdominal fluid collection. Three animals died during the follow-up period due to stroke, hemorrhage, and acute heart failure. After 2 months, a median sternotomy and epicardial echocardiography were performed. Of the surviving 17 animals, 3 developed mild tricuspid regurgitation, 3 developed moderate tricuspid regurgitation, and 11 developed severe tricuspid regurgitation. Eight weeks of pulmonary artery banding resulted in a stable chronic ovine model of right ventricular dysfunction and significant FTR. This large animal platform can be used to further investigate the structural and molecular basis of RV failure and functional tricuspid regurgitation.

Right ventricular failure and functional tricuspid regurgitation are associated with left-sided heart disease and pulmonary hypertension, which contribute significantly to morbidity and mortality in patients. Establishing a chronic ovine model to study right ventricular failure and functional tricuspid regurgitation will help in understanding their mechanisms, progression, and possible treatments.

右心室衰竭和功能性三尖瓣反流的慢性绵羊模型

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to ...

体内心脏移植研究内皮到间充质转化

A Neonatal Heterotopic Rat Heart Transplantation Model for the Study of Endothelial-to-Mesenchymal Transition

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and precludes patients with congenital critical aortic stenosis and hypoplastic left heart syndrome (HLHS) from curative anatomical biventricular surgical repair. Surgical resection is currently the only available therapeutic option, but EFE often recurs, sometimes with an even more infiltrative growth pattern into the adjacent myocardium.
To better understand the underlying mechanisms of EFE and to explore therapeutic strategies, an animal model suitable for preclinical testing was developed. The animal model takes into consideration that EFE is a disease of the immature heart and is associated with flow disturbances, as supported by clinical observations. Thus, the heterotopic heart transplantation of neonatal rat donor hearts is the basis for this model.
A neonatal rat heart is transplanted into an adolescent rat&#39;s abdomen and connected to the recipient&#39;s infrarenal aorta and inferior vena cava. While perfusion of the coronary arteries preserves the viability of the donor heart, flow stagnation within the LV induces EFE growth in the very immature heart. The underlying mechanism of EFE formation is the transition of endocardial endothelial cells to mesenchymal cells (EndMT), which is a well-described mechanism of early embryonic development of the valves and septa but also the leading cause of fibrosis in heart failure. EFE formation can be macroscopically observed within days after transplantation. Transabdominal echocardiography is used to monitor the graft viability, contractility, and the patency of the anastomoses. Following euthanasia, the EFE tissue is harvested, and it shows the same histopathological characteristics as human EFE tissue from HLHS patients.
This in vivo model allows for studying the mechanisms of EFE development in the heart and testing treatment options to prevent this pathological tissue formation and provides the opportunity for a more generalized examination of EndMT-induced fibrosis.

作者聚焦：用于研究内皮-间充质转化的新生儿异位大鼠心脏移植模型  

This work presents an animal model of endothelial-to-mesenchymal transition-induced fibrosis, as seen in congenital cardiac defects such as critical aortic stenosis or hypoplastic left heart syndrome, which allows for detailed histological tissue evaluation, the identification of regulatory signaling pathways, and the testing of treatment options.

新生儿异位大鼠心脏移植模型在内皮间充质转化研究中的应用

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and ...