Development of measurement for hepatic isc hemia-repetfusion in jiry accoidirg to are gulation of activin-follistin system

肝脏缺血再灌注测量的发展

基本信息

批准号：
18591516
负责人：
MORINE Yuji
金额：
$ 2.41万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Background)Ischemia-reperfusion injury to the liver are of clinical importance in humans after hemorrhagic and cardiogenic shock, liver surgery, or liver transplantation. It was reported that blockade of the action of activin with administration of follistatin accelerates recovery from ischemia renal injury by accelerating regeneration after ischemia. In this study we evaluated the role of activin-follistatin on hepatic ischemia reperfusion injury and develop the new measurement for hepatic ischemia reperfusion injury.Method and Results)1) Effects of preactivation by portal vein ligation on liver regeneration (assessment according to activin)This study evaluated the effect of regenerating livers preactivated by PVL following massive hepatectomy. The mRNA expression levels of activin A and ActR II A were significantly higher in regenerating liver group than in normal liver group at late pahse. The regenerating liver preactivated by PVL is restricted late-phase liver regeneration after m … More assive hepatectomy.2) Beneficial Effects of follistatin in hepatic ischemia reperfusion injury Total hepatic ischemia for 30 min was performed followed by reperfusion. Follistatin (1mg/ body) was administered at the time of reperfusion. Four of 5 animals died in control group within 3days after reperfusion, whereas 80% of animals survived in follistatin group (P<0.05). AST was significantly lower at 3 hr after reperfusion in follistatin group (P<0.05). LDH was also lower at 6 hr after reperfusion in follistatin group (P<0.05). Follistatin inhibited the mRNA expression of bA subunit of activin. Moreover the expression of IL-6, which is inflammatory cytokine, was suppressed at 6 hr after reperfusion (P<0.05).3)Beneficial Effects of fluvastatin on Hepatic Ischemia-Reperfusion InjuryRats were divided into 3 groups: fluvastatin group; oral administration of fluvastatin (20mg/kg/day)for 2 days before the operation, control group, sham group. Fluvastatin and control groups were given total hepatic warm ischemia for 30 min. After reperfusion, an expression of mRNA level of endotherial NO synthase (eNOS) and NOX4 was determined by reverse transcriptase (RT)-PCR, tumor necrosis factor- a (TNF-α)and interleukine-1 β(IL-1β) mRNA by real-time RT-PCR. In the fluvastatin group, the expression of eNOS mRNA was significantly higher, and NOX4 mRNA was significantly lower (p<0.05). Furthermore, the expression of inflammatory cytokines mRNA were suppressed in the fluvastatin group. In particular, TNF- α was significantly lower in the fluvastatin group (P<0.05).4) Beneficial Effects of fluvastatin on Liver Microcirculation and Regeneration The liver regeneration rate in fluvastatin group was significantly higher at 72 hours after hepatectomy (p<0.05). Sinusoidal area in fluvastatin group was maintained histologicaly. Furthermore, the expression of-SMA protein in the liver was inhibited in fluvastatin group at 48 hours after hepatectomy.Summary) We were not able to elucidate the regulation of liver regneration using activin, because we had difficulty with the purification of activin. In this study, we clarified the regulatory mechanism of hepatic ischemia-reperfusion injury and effect of several modulators including follistatin for hepatic ischemia-reperfusion injury using molecular biological method. Less

背景）在出血和心源性休克，肝脏手术或肝移植后，对肝脏的缺血 - 再灌注损伤在人类中至关重要。据报道，通过加速缺血后加速再生，加速了卵泡素对卵泡蛋白的作用的阻断，可以加速肾脏肾脏损伤的恢复。在这项研究中，我们评估了激活素炎性蛋白在肝缺血再灌注损伤中的作用，并开发了对肝缺血再灌注损伤的新测量。方法和结果）1）门静脉结扎通过肝脏再生（根据激活的评估）对肝脏再生的影响（根据激活的评估）对效应的效应的效应的影响。重生肝组的激活素A和ACTR II A的mRNA表达水平明显高于Pahse晚期肝组的mRNA表达水平。在M…更多的肝脏肝术后，PVL预体的再生肝脏被限制为后期肝脏再生。2）Follistatin对肝缺血再灌注损伤的有益作用在30分钟内总肝局部缺血总计30分钟。在再灌注时施用Follistatin（1mg/身体）。 5只动物中有4只动物在再灌注后3天内在对照组中死亡，而在Follistatin组中，有80％的动物（p <0.05）存活。卵泡蛋白组再灌注后6小时时，在3小时LDH时的AST显着降低（p <0.05）。 Follistatin抑制了激活素Ba亚基的mRNA表达。此外，在再灌注后6小时（p <0.05）.3）炎症细胞因子的IL-6的表达被抑制在6小时（3）。氟伐他汀对肝缺血 - 再生灌注损伤的有益作用分为3组：Fluvastatin组；手术前2天，对照组，假手术组的口服氟伐他汀（20mg/kg/day）。氟伐他汀和对照组的总肝素温暖缺血30分钟。再灌注后，由逆转录酶（RT）-PCR，肿瘤坏死因子A（TNF-α）和白介碱-1β（IL-1β（IL-1β）mRNA通过实时的RT-PCR确定，由逆转录酶（RT）-PCR，肿瘤坏死因子A（TNF-α）确定MRNA水平的表达。 Fluvastatin组，ENOS mRNA的表达明显更高，NOX4 mRNA明显较低（p <0.05）。此外，在Fluvastatin组中抑制了炎性细胞因子mRNA的表达。特别是，在氟伐他汀组中TNF-α显着降低（p <0.05）.4）氟伐他汀对肝微循环和再生的有益作用肝抑制术后72小时时氟伐他蛋白组的肝脏再生速率显着较低（p <0.05）。在组织学上维持Fluvastatin组的正弦面积。此外，在术后48小时内，在氟伐他汀组中抑制SMA蛋白在肝脏中的表达。我们无法阐明使用激活素对肝脏调节的调节，因为我们难以纯化激活素。在这项研究中，我们阐明了使用分子生物学方法，阐明了肝缺血再灌注损伤的调节机制和几种调节剂的作用，包括肝缺血再灌注损伤的作用。较少的