Name: microsurgical skills of establishing permanent jugular vein cannulation in rats for serial blood sampling of orally administered drug
Uploaded: 2021-12-14T13:00:00
Description: Watch this Scientific Journal Video about Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug at JoVE.com

这项工作由国家自然科学基金（82003692号）资助给张磊。西北工业大学最高学术奖学金授予R. Miao。

以下描述的程序是作为西北工业大学机构动物护理和使用委员会（第202101117号）批准的协议的一部分执行的。
1.术前准备（手术前一天）
注意：所需溶液：生理盐水（0.9%w/v氯化钠），肝素化盐水（1%w/v肝素钠），导管锁溶液，非甾体抗炎药（NSAID），如美洛昔康溶液（2mg / mL）。
<ol><li>溶液制备<ol>
<li>等分试样200μL预包装导管锁定溶液在1.5mL...

<ol>
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K., Niazi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+reliable+method+for+chronic+cannulation+of+the+jugular+vein+for+pharmacokinetic+studies+in+rats.">Simple reliable method for chronic cannulation of the jugular vein for pharmacokinetic studies in rats.</a> Journal of Pharmaceutical Sciences. 72 (9), 1027-1029 (1983).</li><li>Harms, P. G., Ojeda, S. 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+rapid+and+simple+procedure+for+chronic+cannulation+of+the+rat+jugular+vein.">A rapid and simple procedure for chronic cannulation of the rat jugular vein.</a> Journal of Applied Physiology. 36 (3), 391-392 (1974).</li><li>Thrivikraman, K. V., Huot, R. L., Plotsky, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Jugular+vein+catheterization+for+repeated+blood+sampling+in+the+unrestrained+conscious+rat.">Jugular vein catheterization for repeated blood sampling in the unrestrained conscious rat.</a> Brain Research Protocols. 10 (2), 84-94 (2002).</li><li>Weeks, J. R., Davis, J. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+frequent+sampling+of+blood+and+continuous+infusion+of+fluids+in+the+rat+without+disturbing+the+animal.">A method for frequent sampling of blood and continuous infusion of fluids in the rat without disturbing the animal.</a> Physiology &amp; Behavior. 4 (5), 833-836 (1969).</li><li>Terao, N., Shen, D. 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(95), e51881 (2015).</li><li>Karim, N., Ali, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Jugular+vein+cannulation+in+rats+-+A+mini+review.">Jugular vein cannulation in rats - A mini review.</a> Canadian Journal of Pure and Applied Sciences. 3, 929-935 (2009).</li><li>Ling, S., Jamali, F. <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+cannulation+surgery+and+restraint+stress+on+the+plasma+corticosterone+concentration+in+the+rat:+application+of+an+improved+corticosterone+HPLC+assay.">Effect of cannulation surgery and restraint stress on the plasma corticosterone concentration in the rat: application of an improved corticosterone HPLC assay.</a> Journal of Pharmacy &amp; Pharmaceutical Sciences. 6 (2), (2003).</li><li>Lei, 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=Pharmacokinetic+study+of+ellagic+acid+in+rat+after+oral+administration+of+pomegranate+leaf+extract.">Pharmacokinetic study of ellagic acid in rat after oral administration of pomegranate leaf extract.</a> Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences. 796 (1), 189-194 (2003).</li><li>Yan, L. 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=Method+development+and+validation+for+pharmacokinetic+and+tissue+distributions+of+ellagic+acid+using+Ultrahigh+Performance+Liquid+Chromatography-Tandem+Mass+Spectrometry+(UPLC-MS/MS).">Method development and validation for pharmacokinetic and tissue distributions of ellagic acid using Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS).</a> Molecules. 19 (11), 18923-18935 (2014).</li><li>Long, 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=Bioavailability+and+bioactivity+of+free+ellagic+acid+compared+to+pomegranate+juice.">Bioavailability and bioactivity of free ellagic acid compared to pomegranate juice.</a> Food &amp; Function. 10 (10), 6582-6588 (2019).</li><li>Zhang, Y., 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=PKSolver:+An+add-in+program+for+pharmacokinetic+and+pharmacodynamic+data+analysis+in+Microsoft+Excel.">PKSolver: An add-in program for pharmacokinetic and pharmacodynamic data analysis in Microsoft Excel.</a> Computer Methods and Programs in Biomedicine. 99 (3), 306-314 (2010).</li><li>Christakis, 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=Learning+curve+of+vessel+cannulation+in+rats+using+cumulative+sum+analysis.">Learning curve of vessel cannulation in rats using cumulative sum analysis.</a> Journal of Surgical Research. 193 (1), 69-76 (2015).</li><li>Nm, S., Oduola, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Haematological+profile+shows+that+Inbred+Sprague+Dawley+rats+have+exceptional+promise+for+use+in+biomedical+and+pharmacological+studies.">Haematological profile shows that Inbred Sprague Dawley rats have exceptional promise for use in biomedical and pharmacological studies.</a> Asian Journal of Biomedical and Pharmaceutical Sciences. 4 (37), 33-37 (2014).</li><li>Lillie, L. E., Temple, N. J., Florence, L. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reference+values+for+young+normal+Sprague-Dawley+rats:+weight+gain,+hematology+and+clinical+chemistry.">Reference values for young normal Sprague-Dawley rats: weight gain, hematology and clinical chemistry.</a> Human &amp; Experimental Toxicology. 15 (8), 612-616 (1996).</li><li>He, Q. 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=Sex-specific+reference+intervals+of+hematologic+and+biochemical+analytes+in+Sprague-Dawley+rats+using+the+nonparametric+rank+percentile+method.">Sex-specific reference intervals of hematologic and biochemical analytes in Sprague-Dawley rats using the nonparametric rank percentile method.</a> PLoS One. 12 (12), 18 (2017).</li><li>EPA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendations+for+and+Documentation+of+Biological+Values+for+Use+in+Risk+Assessment.">Recommendations for and Documentation of Biological Values for Use in Risk Assessment.</a> U.S. Environmental Protection Agency. , (1988).</li><li>Gaud, N., 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=Single+jugular+vein+cannulated+rats+may+not+be+suitable+for+intravenous+pharmacokinetic+screening+of+high+logP+compounds.">Single jugular vein cannulated rats may not be suitable for intravenous pharmacokinetic screening of high logP compounds.</a> European Journal of Pharmaceutical Sciences. 99, 272-278 (2017).</li><li>Turck, D., 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=Clinical+pharmacokinetics+of+meloxicam.">Clinical pharmacokinetics of meloxicam.</a> Arzneimittel-Forschung/Drug Research. 47 (3), 253-258 (1997).</li><li>Aghazadeh-Habashi, A., Jamali, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacokinetics+of+meloxicam+administered+as+regular+and+fast+dissolving+formulations+to+the+rat:+Influence+of+gastrointestinal+dysfunction+on+the+relative+bioavailability+of+two+formulations.">Pharmacokinetics of meloxicam administered as regular and fast dissolving formulations to the rat: Influence of gastrointestinal dysfunction on the relative bioavailability of two formulations.</a> European Journal of Pharmaceutics and Biopharmaceutics. 70 (3), 889-894 (2008).</li><li>Ludwig, 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=Activation+of+human+cytochrome+P-450+3A4-catalyzed+meloxicam+5+'-methylhydroxylation+by+quinidine+and+hydroquinidine+in+vitro.">Activation of human cytochrome P-450 3A4-catalyzed meloxicam 5 '-methylhydroxylation by quinidine and hydroquinidine in vitro.</a> Journal of Pharmacology and Experimental Therapeutics. 290 (1), 1-8 (1999).</li><li>Zhang, R. X., 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=Nanoparticulate+drug+delivery+strategies+to+address+intestinal+cytochrome+P450+CYP3A4+metabolism+towards+personalized+medicine.">Nanoparticulate drug delivery strategies to address intestinal cytochrome P450 CYP3A4 metabolism towards personalized medicine.</a> Pharmaceutics. 13 (8), (2021).</li></ol>

掌握血管插管技术需要大量的练习，并从每次操作中吸取教训。Christakis等人使用累积总和（CUSUM）分析发现，研究人员需要在一年内练习200只大鼠，然后才能准备对候选药物进行PK评估20只。然而，静脉插管所需的手术时间可以通过执行13，20个大鼠的数量显着减少。使用我们的方案，有效套管颈静脉和收集血液样本的成功率从约50%增加...

从小型实验动物（如啮齿动物、豚鼠和兔子）中反复采集血液样本是药物先导物优化和减少研究中使用的动物数量的重要方面1，2。开发新的诊断工具和药物制剂（例如疫苗）的管道需要获得不同体积的血液，以评估其 在体内的稳健性和性能，例如药代动力学（PK），毒性和敏感性3，4，5。
实验室采集血液样本的方法大致分为两种类型，外科和非手术6。对于研究人员来说，非手术方法相对容易掌握，其中包括常见的技术，例如心脏穿刺，眼眶窦穿刺以及隐静脉和尾静脉出血。通过一些非手术方法可以进行多次血液采样，但样本量很小，会对动物造成身体伤害和心理压力1。另一方面，手术方法是重复静脉穿刺的最爱替代方案，它涉及在动物血管中放置临时或永久性套管7，8，9。在有意识的大鼠中，可以通过套管反复抽取大血容量，同时避免由于处理技术，抑制和麻醉引起的压力和疼痛7，8，10，11。然而，套管植入需要经验丰富的研究人员进行足够的培训，才能成功收集血液。
通过大鼠颈静脉插管（JVC）收集血液是研究药物PK6，10，12，13的最广泛使用的方法。然而，JVC大鼠模型的建立需要仔细练习显微外科技能和术后护理和维护知识。特别是，手术后，大鼠需要给予镇痛药和足够的恢复时间才能达到稳定的生理状态，用于进一步的实验13，14，15。虽然体重增加（即>10g）是大鼠恢复的有效且常用的指标，但大鼠由于脱水，感染和炎症而在术后意外死亡并不罕见，这在早期发病时可能很微妙地注意到14，15。此外，JVC模型中的导管阻塞仍然是采血过程中的一个问题。
本方案已详细演示了麻醉大鼠JVC的显微外科手术，特别关注颈静脉的识别，分离和插管。强调了在恢复阶段对大鼠进行生理和血液学监测的重要性。最后，通过静脉导管收集连续血液样本，研究口服生物利用度差（即全身浓度低）的天然酚鞣花酸的PK，以验证JVC大鼠模型。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.5 mL test tube containing EDTA anticoagulant</td>
			<td>Xinkang</td>
			<td>N/A</td>
			<td>collecting blood samples for hematology test</td>
		</tr>
		<tr>
			<td>0.5*20 mm 1.0-mL syringe</td>
			<td>KLMEDICAL</td>
			<td>N/A</td>
			<td>washing or replacing the fluid with saline</td>
		</tr>
		<tr>
			<td>0.6*28.5 mm 5.0-mL syringe</td>
			<td>HD</td>
			<td>N/A</td>
			<td>Subcutaneous injection</td>
		</tr>
		<tr>
			<td>1.0-mL Blunt tipped syringe (22G)</td>
			<td>skillsmodel</td>
			<td>S4-PKT22G</td>
			<td>Inject the saline and collect blood samples through catheter</td>
		</tr>
		<tr>
			<td>1.5 mL sterile microcentrifuge tube</td>
			<td>Axygen</td>
			<td>MCT-150-C-S</td>
			<td>Store sterile catheter lock solution heparinized saline and meloxicam solution</td>
		</tr>
		<tr>
			<td>1.5&#160;mL microcentrifuge tubes</td>
			<td>Biosharp</td>
			<td>BS-15-M</td>
			<td>blood collection</td>
		</tr>
		<tr>
			<td>1/2&#160; circle cutting 5*12 mm suture needle</td>
			<td>skillsmodel</td>
			<td>S4-FHZ</td>
			<td>Thread the muscle layer to fix the catheter</td>
		</tr>
		<tr>
			<td>3/8 circle cutting 7*17 mm suture needle</td>
			<td>skillsmodel</td>
			<td>S5-FHZ</td>
			<td>Suture the incision of rat cortex</td>
		</tr>
		<tr>
			<td>6-0 sterile non-absorbable silk suture thread</td>
			<td>JUNSHENG</td>
			<td>N/A</td>
			<td>ligature</td>
		</tr>
		<tr>
			<td>75% medical alcohol</td>
			<td>HONGSONG</td>
			<td>N/A</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>Adhensive tape</td>
			<td>LIUTAI</td>
			<td>N/A</td>
			<td>positioning the rat</td>
		</tr>
		<tr>
			<td>Autoclave sterilization tape</td>
			<td>Biosharp</td>
			<td>BS-QT-028</td>
			<td>Mark sterilized items</td>
		</tr>
		<tr>
			<td>Automated blood cell counter</td>
			<td>Sysmex</td>
			<td>XN-550</td>
			<td>Hematology test</td>
		</tr>
		<tr>
			<td>Castroviejo micro scissors</td>
			<td>skillsmodel</td>
			<td>WA1010</td>
			<td>Cut the opening in the blood vessel</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Thermo Fisher Scientific</td>
			<td>75002402</td>
			<td>Plasma preparation</td>
		</tr>
		<tr>
			<td>Clean cushion</td>
			<td>Qingjie</td>
			<td>N/A</td>
			<td>Prepare the operation area</td>
		</tr>
		<tr>
			<td>Cotton balls</td>
			<td>HC</td>
			<td>N/A</td>
			<td>Wound disinfection and sterilization</td>
		</tr>
		<tr>
			<td>Cotton swabs</td>
			<td>BEITAGOGO</td>
			<td>N/A</td>
			<td>Disinfection</td>
		</tr>
		<tr>
			<td>Curved hemostat</td>
			<td>skillsmodel</td>
			<td>N/A</td>
			<td>ligature</td>
		</tr>
		<tr>
			<td>DN50 Stainless-steel rat restrainer</td>
			<td>skillsmodel</td>
			<td>S4-RGDQ1</td>
			<td>Restrict the movement of rats for easy operation</td>
		</tr>
		<tr>
			<td>Ellagic acid</td>
			<td>Aladdin</td>
			<td>E102710-25g</td>
			<td>natural phenol for oral administration</td>
		</tr>
		<tr>
			<td>Half-curved forceps</td>
			<td>skillsmodel</td>
			<td>53072</td>
			<td>Lift the muscle layer and tissue, isolate the jugular vein and tie the suture</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Warm mate</td>
			<td>N/A</td>
			<td>preventing heat loss of animal</td>
		</tr>
		<tr>
			<td>Heparin sodium</td>
			<td>Solarbio</td>
			<td>H8060</td>
			<td>anticoagulant</td>
		</tr>
		<tr>
			<td>Iodophor</td>
			<td>Xidebao</td>
			<td>N/A</td>
			<td>Clean the wound</td>
		</tr>
		<tr>
			<td>Iris scissors</td>
			<td>skillsmodel</td>
			<td>54002</td>
			<td>Bluent separation the muscle layer</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD</td>
			<td>R510-22-16</td>
			<td>anaesthesia</td>
		</tr>
		<tr>
			<td>LED lamp</td>
			<td>EMPERORFEEL</td>
			<td>N/A</td>
			<td>sugery</td>
		</tr>
		<tr>
			<td>Liquid chromatography-mass spectroscopy</td>
			<td>Thermo Fisher Scientific</td>
			<td>VQF01-20001/ TSQ02-10002</td>
			<td>detection of drug concentration in plasma</td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>Hongqiang</td>
			<td>N/A</td>
			<td>Analgesic</td>
		</tr>
		<tr>
			<td>Normal saline</td>
			<td>KL</td>
			<td>N/A</td>
			<td>Prepara the solution and protect blood vessels from drying out</td>
		</tr>
		<tr>
			<td>Pet razor</td>
			<td>Codos</td>
			<td>3180</td>
			<td>Shaving the fur</td>
		</tr>
		<tr>
			<td>Phosphate-buffered saline</td>
			<td>ZHHC</td>
			<td>PW012</td>
			<td>Preparation of Ellagic acid solution</td>
		</tr>
		<tr>
			<td>PU catheter</td>
			<td>skillsmodel</td>
			<td>RJVC-PU</td>
			<td>Jugular vein cannulation</td>
		</tr>
		<tr>
			<td>Small animal operation anesthesia console</td>
			<td>RWD</td>
			<td>68620</td>
			<td>Operation workstation</td>
		</tr>
		<tr>
			<td>Spray bottle</td>
			<td>Other</td>
			<td>N/A</td>
			<td>aseptic workstation</td>
		</tr>
		<tr>
			<td>Stainless steel plug (22G)</td>
			<td>skillsmodel</td>
			<td>S4-PKD22G</td>
			<td>Plug the catheter to ensure its sealing</td>
		</tr>
		<tr>
			<td>Stainless steel trochar</td>
			<td>skillsmodel</td>
			<td>S$-PKDGZ</td>
			<td>Guide the catheter exteriorization</td>
		</tr>
		<tr>
			<td>Sterile lock solution</td>
			<td>skillsmodel</td>
			<td>SK-FB</td>
			<td>lock the catheter to ensure its sterility</td>
		</tr>
		<tr>
			<td>Straight feeding needle</td>
			<td>skillsmodel</td>
			<td>N/A</td>
			<td>Oral gavage</td>
		</tr>
		<tr>
			<td>Surgical pouch</td>
			<td>BKMAM</td>
			<td>N/A</td>
			<td>container for sterilization of surgical instruments</td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td>skillsmodel</td>
			<td>J21070</td>
			<td>Cut incision on rat skin</td>
		</tr>
		<tr>
			<td>Vessel dilator balanced forceps</td>
			<td>skillsmodel</td>
			<td>WA3020</td>
			<td>Expand the blood vessel and guide the cannula to slide in</td>
		</tr>
		<tr>
			<td>ZS-MV Small animal anesthesia machine</td>
			<td>ZSLab</td>
			<td>1057003</td>
			<td>inducing and maintaining anaesthesia</td>
		</tr>
	</tbody>
</table>

microsurgical skills of establishing permanent jugular vein cannulation in rats for serial blood sampling of orally administered drug

小型实验动物的血液采样对于药物先导物优化是必要的，但会对实验动物造成巨大的伤害和压力，这可能会影响结果。大鼠的颈静脉插管（JVC）是一种广泛使用的重复采血模型，但需要对手术技能和动物护理进行充分的培训。本文详细介绍了用于建立和维护永久性JVC大鼠模型的显微外科手术程序，特别关注颈静脉套管的放置和密封。监测生理（例如，体重，食物和水摄入量）和血液学参数的重要性，在大鼠恢复期间在手术后6天内提供结果得到强调。在JVC大鼠模型中测定口服天然酚鞣花酸的药物 - 血浆浓度 - 时间曲线。

该协议已彻底展示了如何使用显微外科技能建立长期JVC模型以进行连续采血。 图1A 显示了用于进行手术的基本手术器械和材料。还说明了具有三个蓝色标记的PU导管的规格，这有助于指导研究人员在步骤3.3中放置静脉套管，如何使用PU导管上的标记来引导插管（图1B）。同样重要的是要了解建立JVC大鼠模型所需的时间表（图1C</stron...

Watch this Scientific Journal Video about Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug at JoVE.com

Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug

详细的显微外科技术被证明可以建立一个长期的颈静脉插管大鼠模型，用于同一动物的序贯血液采集。在大鼠的恢复阶段监测生理和血液学参数。该模型已应用于研究口服多酚的药代动力学，而不会诱导动物应激。

建立大鼠永久性颈静脉插管的显微外科技能，用于口服药物的连续血液采样

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, ...

从小型实验动物中重复取样血液是药物先导物优化的一个重要方面。该协议详细介绍了建立永久性颈静脉插管大鼠模型的显微外科技能。该技术可以有效地建立颈静脉插管大鼠模型，并在避免压力和疼痛的同时，通过插管反复收集有意识的大鼠的血液样本。定位颈静脉及其插管非常重要，因为此模型无法直观地演示解剖复杂性。首先，用75%医用酒精制备无菌工作站，然后用与氧气混合的异氟醚麻醉剃须的大鼠。皮下注射美洛昔康溶液，剂量为每公斤2毫克，然后用镊子小心地抬起颈部中线右侧锁骨附近的皮肤，并使用一把手术剪刀，向胸部开一个1.5至2厘米长的切口。为了暴露颈静脉下方，使用虹膜剪刀钝化薄薄的组织覆盖物。颈外静脉的头端近端由两个分支组成，可以直观地识别。抬起颈静脉及其结缔膜组织，以可视化附着在颈静脉上的淋巴腺。小心地将沿血管方向的静脉与周围的肌肉、脂肪和其他组织分开。在不损伤侧支血管的情况下将镊子轻推颈静脉下方，并在静脉下通过两块6-0缝合线，分别标记血管的两端。将一块缝合线尽可能向大鼠头拉，并用镊子用两到三个结将静脉颅骨缝合。将第二个结扎带放在静脉的尾端，用一个松散的结。将11厘米的聚氨酯或PU导管连接到装有肝素化盐水的钝头注射器上，然后缓慢将肝素化的盐水推入导管中以避免气泡。将颈静脉下方镊子的非尖端平坦侧轻推到另一侧退出。用一对卡斯特罗维耶霍微剪刀在颅带附近的静脉上做一个小的V形切口，并用肘部血管扩张器镊子的尖端轻轻打开切口。切除颈静脉导管前端的斜开口。用镊子植入管的斜端，并将其滑入颈静脉。在推进导管时，慢慢拔出肘部显微外科镊子，并用镊子夹住血管的外表面。在击中PU管的第一个蓝色标记时停止插入导管，该标记的长度约为三厘米。使用镊子将插入的导管固定在静脉上，同时使用尾部和喙状结扎。使用缝合针将6-0缝合线穿过切口右侧暴露的组织，并用止血剂绑扎结扎。然后在第二个蓝色标记处弯曲导管以使用相同的结扎线，并避免阻塞PU管。剪下所有多余的缝合线后，用22号不锈钢塞替换钝头注射器，关闭导管。将大鼠置于背部位置，并用浸泡在75%医用酒精中的棉球轻轻清洁肩胛骨之间的区域。使用手术剪刀，在背颈中心做一个非常小的切口。通过背侧切口，引导并轻轻地将皮肤下方的推杆推向颈部右侧的腹侧切口。将静脉导管放入穿刺器中，然后拔出并将静脉导管引导至背侧切口。将外切导管固定到肌肉层后，用6-0尼龙缝合线和缝合针关闭腹侧和背侧切口的皮肤层，并用碘伏法拭子所有手术切口。接下来，用指尖扣住导管，取下导管塞。放置一个新的钝头注射器，然后慢慢拉回注射器以测试血流。用指尖再次握住导管。使用钝头注射器将0.2毫升肝素盐水和0.1毫升锁溶液注入导管，然后用不锈钢塞子代替注射器。松开导管并稍微推入插头，以确保导管的密封性。收集血液样本进行血液学检查，将大鼠置于抑制器中，打开塞子，然后将注射器插入静脉PU导管中，以确保导管不被阻塞。丢弃最初抽出的含有血液，肝素化盐水和导管锁紧溶液混合物的血液。使用新的注射器，收集150微升新鲜血液样本，并将血液样本转移到先前喷有K2EDTA的0.5毫升管中。注射150微升预热的生理盐水，并通过导管输注0.2毫升无菌肝素化生理盐水。将100微升锁定溶液注入导管中，以确保在下一次样品收集之前导管的密封性和无菌性。在目前的研究中，研究了术后六天的生理和血液状况。大多数大鼠在手术后四至六天内恢复，表现为体重增加超过10克，定期进食，与感染，脱水和炎症有关的选定血液成分，包括白细胞，红细胞，血红蛋白和血小板计数。因此，大鼠在手术后需要至少四到六天的恢复时间。大量饮水表明术后第一天脱水。此外，在已建立的颈静脉插管大鼠模型中研究了天然多酚鞣花酸的药代动力学，其生物利用度差表现为其24小时内血浆浓度低。正确隔离颈静脉是最重要的第一步，因为嵌入其他软组织的颈静脉可能不会立即被研究人员看到。同一动物内的序贯血液采样可以在已建立的JVC大鼠模型中进行。它是评估药物配方体内性能的有用且强大的工具。

Research

JoVE Journal

Medicine

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

科研

医学

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

培养一个精密的显微外科技术在大鼠颈总动脉，颈外静脉移植：介入

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

成年小鼠静脉高压型号：颈总动脉，以颈外静脉吻合。

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System

无麻醉大鼠尾静脉血采集系统的研究

Blood can be collected from rats in a number of sampling locations. For instance, the tail vein is a superior location for blood sampling. However, the ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

颈动脉介入移植中颈静脉持久性转基因表达的家兔模型

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

锁骨下静脉穿刺作为大鼠采血的一种替代方法

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...

Blood Collection Through Subclavian Vein Puncture in Mice

小鼠锁骨下静脉穿刺采血

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in ...

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...

Comparative Study of Blood Collection Techniques in Rats

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Context Dependent Blood Collection Methods in Rats (Video) | JoVE

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood ...