DEVELOPMENT OF SLIT LASER MICROSCOPE FOR QUANTIFICATION OF TISSUE PO2 DISTRIBUTION

开发用于量化组织 PO2 分布的狭缝激光显微镜

• 批准号: 09555120
• 负责人: SHIBATA Masahiro
• 金额: $ 4.93万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09555120/
• 关键词: OXYGEN TENSION; VITAL MICROSCOPY; LASER; MICROCIRCULATION; Pd-PORPHYRIN; SKELETAL MUSCLE; Pdポルフィリン; リン光寿命; Stern-Volmer

To clarify the oxygen transport across the microvessel and oxygen distribution in the skeletal muscle, we have newly designed an intravital laser microscope with oxygen-dependent quenching of phosphorescence technique. The phosphorescence probe for oxygen was injected into the systemic blood, and phosphorescence excited by N_2/dye pulse laserin the tissue area of 15 mum in diameter was measured by a photomultiplier. In vitro and in vivo oxygen tension (pO2) measurements revealed the efficiency of this method because of its non-invasiveness, accuracy, and rapid response time. This technique was applied to pO2 measurements in the rat microcirculation. The arteriolar and venular pO2 measurements were performed at the several regions classified by branching order. The mean arteriolar pO2 of different orders decreased according to increase the branching(A1 : 72.4, A2 : 51.8 and A3 : 44.1 mmHg), however venular pO2 were independent on vessel orders(V1 : 27.5, V2 : 28.6 and V3 : 32.0 mmHg). Interstitial pO2 adjacent to arterioles of different orders were significantly lower than those of intravascular pO2 (I1 : 47.8, I2 : 35.8 and I3 : 24.4 mmHg). In conclusion our new designed intravital microscope was proved the effective method to clarify the mechanisms of oxygen transport across the microvessels and the oxygen distribution in the skeletal muscle. In addition, the existence of significant reduction of pO2 levels in arterioles of different orders and large pO2 gradients at the interface between blood and tissue suggest that oxygen consumption of the smooth muscle and/or endothelium in the skeletal muscle arteriole might be high.

为了阐明氧气在微血管中的运输和骨骼肌中的氧气分布，我们新设计了一种具有氧依赖性磷光猝灭技术的活体激光显微镜。将氧磷光探针注入全身血液中，用光电倍增管测量直径15μm的组织区域内N_2/染料脉冲激光激发的磷光。体外和体内氧分压 (pO2) 测量揭示了该方法的效率，因为其无创性、准确性和快速响应时间。该技术应用于大鼠微循环的 pO2 测量。在按分支顺序分类的几个区域进行小动脉和静脉 pO2 测量。不同阶次的平均小动脉pO2随着分支的增加而降低（A1：72.4，A2：51.8和A3：44.1 mmHg），而小静脉pO2与血管阶次无关（V1：27.5，V2：28.6和V3：32.0 mmHg）。不同级次小动脉附近的间质 pO2 均显着低于血管内 pO2（I1：47.8、I2：35.8 和 I3：24.4 mmHg）。总之，我们新设计的活体显微镜被证明是阐明氧气跨微血管运输机制和骨骼肌中氧气分布的有效方法。此外，不同级别的小动脉中 pO2 水平显着降低以及血液和组织界面处的大 pO2 梯度的存在表明骨骼肌小动脉中平滑肌和/或内皮的耗氧量可能较高。

项目成果

N.Satake, M.Shibata, S.Shibata: "The involvement of K_<Ca'> K_<ATP> and K_V channels in vasorelaxing responses to acetylcholine in rat aortic rings." Gen.Pharmacol.28. 453-457 (1997)

N.Satake、M.Shibata、S.Shibata：“K_<Ca>、K_<ATP> 和 K_V 通道参与大鼠主动脉环中乙酰胆碱的血管舒张反应。”

S.Ichioka, M.Iwasaka, M.Shibata, K.Harii, A.Kamiya, S.Ueno: "Biological effects of magnetic fields on the microcirculatory blood flow in vivo : A preliminary report." Med.& Biol.Eng.& Comput.36. 91-95 (1998)

S.Ichioka、M.Iwasaka、M.Shibata、K.Harii、A.Kamiya、S.Ueno：“磁场对体内微循环血流的生物效应：初步报告。”

T.Takahasih, M.Shibata, A.Kamiya: "Mechanism of macromolecule concentration in collecting lymphatics in rat mesentery." Microvasc.Res.54. 193-205 (1997)

T.Takahasih、M.Shibata、A.Kamiya：“大鼠肠系膜淋巴管收集中大分子浓度的机制”。

M.Shibata,A.Kamiya: "A phosphorescence measurement of tissue oxygen distribution in the rat cremaster muscle." Proc.Eur.Microcirc.177-182 (1998)

M.Shibata，A.Kamiya：“大鼠提睾肌中组织氧分布的磷光测量。”

N.Satake,M.Shibata,S.Shibata: "The involvement of K_<Ca>,K_<ATP> and K_ν channels in vasorelaxing responses to acetylcholine in rat aortic rings." Gen.Pharmacol.28. 453-457 (1997)

N.Satake、M.Shibata、S.Shibata：“K_<Ca>、K_<ATP> 和 K_ν 通道参与大鼠主动脉环乙酰胆碱的血管舒张反应。”Gen.Pharmacol.28（1997 年） ）