摘要
目的:探讨细胞穿透肽PEP-1介导的血红素加氧酶-1(HO-1)对大鼠肝脏缺血再灌注损伤(HIRI)的保护作用。方法:(1)用基因工程学方法人工合成融合蛋白PEP-1-HO-1。(2)选择雄性SD大鼠,随机分为三组(n均=8):假手术组(S组)只开腹,不予干预。HIRI模型组(HIRI组),采用夹闭肝动脉和门静脉30min后恢复血流;HIRI+PEP-1-HO-1预处理组(HIRI+HO-1组),夹闭肝动脉和门静脉前经门静脉注射PEP-1-HO-1蛋白1mg,其余处理同HIRI组。(3)实验完成后,取三组大鼠下腔静脉血,采用自动生化分析仪测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、γ-谷氨酰转肽酶(γ-GT)、总胆红素(TBIL)等肝功能指标。处死大鼠,取部分肝脏进行组织切片、HE染色,观察各组肝组织病理学变化。结果:成功制备高纯度PEP-1-HO-1融合蛋白。HIRI组大鼠各项肝功能指标均显著高于S组(P<0.01),HIRI+HO-1组各项肝功能指标值明显低于HIRI组(P<0.01),但仍高于S组(P<0.05)。HIRI组肝细胞肿大或呈球形,胞浆疏松水样变或完全透明,伴炎细胞浸润,可见片状坏死。HIRI+HO-1组肝脏损伤程度较HIRI组明显改善,炎性细胞浸润及肝细胞坏死程度明显减轻,但与S组比较,肝脏组织损伤仍明显。结论:用细胞穿透肽PEP-1介导HO-1预处理能有效保护肝细胞,明显减轻肝功能损害。
Objective： To study liver protection of PEP-1 mediated heme oxygenase 1（HO-1） in rat hepar ischemia-reperfusion injury（HIRI）. Method： （1） Synthetic protein （PEP-1-HO-1） was made by genetic engineering methods. （2）24 male SD rats were randomly assigned to three groups：the sham group（S group）, HIRI model group （HIRI group）, HIRI and PEP-1-HO-1 pretreatment group（HIRI＋ HO-1 group）. （3） After experiment completely, extraction of three groups of inferior vena venous blood of rats by automatic biochemical analyzer determine serum： cereal third transominase （ALT）, aspertate aminotransferase （AST）, gamma glutamyl transpeptidase（γ-GT）, total bilirubin （TBIL） and liver function indicators. Taking part executed in rats after the liver biopsy HE staining,each group liver tissue pathological changes were observed. Results： Successful preparation of high purity PEP-1-HO-1 fusion protein. Rats liver function index HIRI group were significantly higher than S group （P〈0. 01）, HIRI＋ HO-1 set of various liver function index was lower than that in group HIRI （P〈0.01） ,but it was still higher than the group S （P〈0.05）. HIRI＋ HO-1 set of HIRI group significantly reduced degree of liver injury, inflammatory cell infiltration and necrosis of liver cells was significantly reduced,but compared with the group S , the liver tissue damage was still obvious. Conclusion： It can pretreatment effectively protect the liver cells with cell penetrating pep- tides PEP-1 mediated heme oxygenase 1 ,significantly reduce liver damage.
出处
《微循环学杂志》
2015年第1期14-17,F0003,共5页
Chinese Journal of Microcirculation
基金
湖北省教育厅科学技术研究计划重点项目(编号D20082045)
