Studies of non-investigate evaluation and development of therapy for ischemic injury of the liver using new technic of near-infrared specroscopy

应用近红外光谱技术（NIR）检测大鼠肝移植术后一氧化氮（NO）的合成和组织氧合，以监测肝移植术后的排斥反应和缺血性损伤。1.对受者移植物进行近红外光谱分析，获得亚硝基血红蛋白、氧合血红蛋白和氧化型细胞色素氧化酶（Cyt.aa_3）的相对变化。未处理的同种异体移植组从急性排斥反应的早期就检测到亚硝基血红蛋白水平的显著升高，脱氧精胍灵治疗可以抑制这种升高。同基因肝移植后长期冷保存至第3天，NO合成也增加。我们的近红外光谱方法具有检测组织中超过10 μ M NO水平的NO的灵敏度<-4>，这是通过使用自发NO吸收器（FK 409）的体内测定来分析的。与同基因组相比，未处理的同种异体组中的oxy-Hb和氧化的Cyt.aa_3的水平显着降低时，实质疾病被证实组织学上，保持不变。因此，我们的近红外技术被证明是能够同时评估免疫反应和免疫诱导的破坏程度的肝移植后，通过监测NO的合成和组织氧合。2.我们开发了一种含有丝氨酸蛋白酶抑制剂甲磺酸萘莫司他（NM）的新冲洗液，NM冲洗和Kuppfer细胞阻断在预防冷保存肝移植缺血性损伤中发挥协同作用。我们还报道了禁食大鼠的肝脏移植后获得对缺血性损伤的耐受性。我们检测了进入禁食大鼠腹水的热休克蛋白（GRP 78，HPS 70）和TNF α片段被显著抑制。这些结果表明，诱导肝细胞保护物质可能有助于对缺血损伤的耐受。

Near-infrared spectroscopy (NIRs) was applied to rat liver for assessing nitric oxide (NO) synthesis and tissue oxygenation as a means of monitoring the rejection response and ischemic injury following liver transplatation. 1. NIRs was performed on the grafts in recipients, and the relative changes in nitrosyl-Hb, oxy-Hb, and oxidized cytochrome oxidase (Cyt.aa_3) were obtained. A significant rise in the level of nitrosyl-Hb was detected from the early phase of acute rejection in the untreated allogeneic group, and the increase was suppessed by deoxy-spergualin treatment. NO synthesis was also increased after long cold preservation in syngeneic liver transplants until day 3. Our NIRs method have a sensitivity to detect NO in the tissues at the level over 10^<-4>M NO,which was analyzed by use of in vivo assay using a spontaneous NO releaser (FK409). Levels of oxy-Hb and oxidized Cyt.aa_3 in the untreated allogeneic group were significantly decreased when parenchymal disorder was confirmed histologically, compared with those in syngeneic group, which remained constant. Thus, our NIRs technique was shown to be capable of assessing simultaneously both the immune resoponse and the degree of immune-induced destruction of graft tissue following liver transplantation through monitoring of NO synthesis and tissue oxygenation. 2. We developed a new rinse solution containing nafamostat mesilate (NM) (a serine protease inhibitor), and NM rinse and Kuppfer cell blockade exert synergistic effect in preventing ischemic injury in cold-preserved liver transplantation. We also reported that livers from fasted rats acquire tolerance to ischemic injury following transplantation. We examined heat shock protein (GRP78, HPS70) and TNF alpha section into the ascitic fluid of fasted rats is markedly suppressed. These findings suggest that induction of cytoprotective substances in the liver may contribute to the tolerance to ischemic injury.