“体外骨骼肌微血管制备离体血管反应性的调查
Summary
一个<em>体外</em>编制说明两个血管反应性血管活性物质的刺激和通过被动墙力学的基本结构性能评估的审讯股薄肌的最大阻力小动脉的隔离。
Abstract
隔离的微血管准备是体外制备，允许考试因素的控制血管直径的不同贡献，因此，灌注阻力1-5。这是一个经典的实验准备，在很大程度上，最初是由内田等 15，几十年以前。这最初的描述提供了进行了广泛的修改和提高，主要是在实验室的布赖恩博士杜陵在弗吉尼亚6-8大学，技术的基础上，我们提出了电流的方法，在下面的网页。这种准备，具体是指在大鼠作为首选微血管纤细的动脉，但基本的准备，可以很容易地应用到几乎任何其他组织或器官9-13跨物种隔离的船只。机械（即二维）在孤立的微血管的变化，可以很容易地评估在浩如烟海的生理（例如，缺氧，血管内的压力，或剪切）或药理的挑战，并能提供洞察到机械元素组成一个完整的综合措施，虽然体外 ，组织响应。这种方法的意义是，它允许简便操作的影响微血管直径的综合调控，同时也让许多从其他来源，包括血管内压力（生肌），自主支配，血流动力学的贡献控制（例如，剪切应力），内皮依赖或独立的刺激，激素，实质影响，提供了部分清单。在适当的实验条件和适当的目标，这可以作为一个优势在体内或原位组织/器官的筹备工作，不轻易让浅显的控制更广泛的系统性变量。
马约旦基本上是该制剂的限制其优势的结果。根据定义，这些船只的行为被许多最重要的贡献者调节血管阻力已被删除，包括神经，体液免疫，代谢，因此等的条件下，研究，研究者应注意避免过度所收集的数据利用这个准备的解释和推断。该制剂方面关心的其他重要领域，它可以很容易损坏，如内皮衬里或血管平滑肌细胞成分，如变量的错误源可以被引入。强烈建议，个别研究者利用，以确保该制剂的质量，无论是在实验开始，并定期在整个协议的过程中适当的测量。
Protocol
Discussion
提出的协议描述的骨骼肌微血管隔离，清除和双插管，虽然这一般技术可以很容易地适用于大多数组织。对于当前的手稿，“动脉”一词已被用于由作者来形容一个阻力血管直径在70-120微米休息积极的基调下，这也是一个主要因素调节器官灌注阻力或之间不等组织。
一些修改，这个系统可以安装多个应用程序的具体调查，并随时可以提供更深入的实验（例如，17膜电?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是由美国心脏协会（EIA 0740129N）和NIH的T32 HL90610支持。
Materials
| Reagents and Equipment | Company | Comments/Catalogue # |
| Vessel Chamber | Custom | Dave Eick (MCW) |
| Heated Circulating Water Bath | PolyScience and Haake | Haake DC 10 |
| Pipets | Frederick Haer & Co. | Capillary Tubing 2.0 mm OD x 1.0 mm ID (27-33-1) |
| Pressure Monitor | World Precision Instruments | |
| Water Jacketed Reservoir | Custom | |
| External Light Source | World Precision Instruments | Novaflex |
| Pipet Puller | MicroData Instruments | PMP102 Micropipet Puller |
| Full complement of surgical tools | Fine Science Tools | Dumont |
| Ultra Fine Forceps | Fine Science Tools | Inox #5 |
| Silk Suture Thread | Ethilon | #10-0 or 9-0 |
| Stereo Microscope | Olympus | Olympus SZ-11 |
| Analog Video Calipers | Boeckeler | Via Controller (Via-100) |
| High Resolution Analog Camera | Panasonic | GP-MF 602 |
| Oxygen Tank | Regional | 21% balance nitrogen and 5% CO2 balance nitrogen |
| Tubing | Tygon | |
| Drain Pump | Cole Parmer Instrument Co. | |
| Modified Rat PSS | See recipe below | |
| Van Breemen’s Relaxant PSS | See recipe below |
Table 1. A list of the major components of isolated microvessel station setup presented in the Figures.
| Modified Rat PSS Recipe | To make two liters of PSS | 20X Salt Stock (2L) | 20X Buffer Stock (2L) |
| NaCl | 278.0 g | ||
| KCl | 14.0 g | ||
| MgSO4-7H2O | 11.5 g | ||
| CaCl2-H2O | 9.4 g | ||
| NaHCO3 | 80.8 g | ||
| EDTA | 0.4 g | ||
| NaH2PO4 | 0.28 g | ||
| Glucose | 1.98 g | ||
| 20x Salt Stock | 100 mL | ||
| 20x Buffer Stock | 100 mL | ||
| Distilled Water | 1800 mL |
Table 2. Recipe for standard physiological salt solution (PSS) used in the isolated microvessel protocols.
Comments on Recipe: Make 2 L of Salt Stock and 2 L of Buffer Stock. These can be refrigerated when not being used, but shake them well and often before preparing PSS. The additional ingredients are added at the time of preparation of final PSS.
| Van Breemen’s Relaxant PSS | To make 2 liters of PSS | 20X Salt Stock (1L) | 20X Buffer Stock (1L) |
| NaCl | 107.4 g | ||
| KCl | 7.0 g | ||
| MgSO4-7H2O | 5.76 g | ||
| MgCl2-6H2O | 81.32 g | ||
| NaHCO3 | 40.4 g | ||
| EDTA | 0.2 g | ||
| EGTA | 15.22 | ||
| NaH2PO4 | 0.28 g | ||
| Glucose | 1.98 g | ||
| 20x Salt Stock | 100 mL | ||
| 20x Buffer Stock | 100 mL | ||
| Distilled Water | 1800 mL |
Table 3. Recipe for Van Breemen’s relaxant physiological salt solution (PSS) used in the isolated microvessel protocols under conditions of zero active tone.
Comments on Recipe: Make 1 L of Salt Stock and 1 L of Buffer Stock. These can be refrigerated when not being used, but shake them well and often before preparing PSS. The additional ingredients are added at the time of preparation of final relaxant PSS.
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