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（NIRs）技术对大鼠肝脏进行一氧化氮（NO）合成和组织氧合评价，作为监测肝移植术后排斥反应和缺血性损伤的手段。1. 对受体移植物进行近红外光谱分析，得到亚硝基- hb、氧基- hb和氧化细胞色素氧化酶（Cyt.aa_3）的相对变化。未经治疗的同种异体组急性排斥反应早期检测到亚硝基- hb水平显著升高，脱氧精巴林治疗抑制了这种升高。同基因肝移植长时间冷保存至第3天，一氧化氮合成也增加。我们的近红外光谱方法对组织中NO的检测灵敏度超过10^<-4>M NO，使用体内实验，使用自发NO释放剂（FK409）进行分析。氧合血红蛋白和氧化细胞的水平。与同基因组相比，同种异体治疗组在组织学上确诊实质疾病时，Aa_3含量明显降低，并保持不变。因此，我们的近红外光谱技术被证明能够通过监测NO合成和组织氧合来同时评估肝移植后移植组织的免疫反应和免疫诱导破坏程度。2. 我们研制了一种含有甲磺酸纳莫他酯（NM）（一种丝氨酸蛋白酶抑制剂）的新型冲洗液，NM冲洗液和kupfer细胞阻断剂在预防冷保存肝移植缺血损伤中发挥协同作用。我们还报道了禁食大鼠的肝脏在移植后获得对缺血性损伤的耐受性。我们检测热休克蛋白（GRP78、HPS70）和肿瘤坏死因子α切片进入禁食大鼠腹水是否明显抑制。这些发现提示肝脏细胞保护物质的诱导可能有助于对缺血性损伤的耐受。

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.