摘要
目的:探讨组织蛋白酶S抑制剂(Cat S-I)影响小鼠缺血诱导的血管新生的机制。方法:将8周龄野生型小鼠(C57/BL6)随机分为两组:对照组(n=20)采用基础饲料饲养基础上再注射5%羧甲基纤维素钠盐,Cat S-I组(n=20)在基础饲料饲养基础上再注射Cat S-I[1 mg/(kg·d)],分别每天1次,共17天。腹腔注射3天后建立下肢缺血模型。用激光多普勒超声血流仪检测下肢血流,计算缺血肢与非缺血肢血流面积比;术后第4天利用免疫蛋白印迹(Western)法检测过氧化物酶增殖物激活受体-γ(PPAR-γ)、磷酸化蛋白激酶B(p-Akt)、磷酸化内皮型一氧化氮合酶(p-e NOS)和血管内皮生长因子(VEGF)等蛋白水平;术后第7天,取缺血骨骼肌作冰冻切片,采用免疫组织化学方法测定毛细血管密度。结果:(1)Cat S-I明显抑制下肢血流的恢复。缺血与非缺血下肢血流比值统计结果显示:Cat S-I组小鼠缺血下肢血流恢复明显慢于对照组(P<0.05)。(2)Cat S-I组明显抑制毛细血管的形成。术后第14天非缺血骨骼肌中,与对照组比较,Cat S-I组毛细血管形成略减少,但差异无统计学意义(P>0.05);但在缺血骨骼肌中,Cat S-I组小鼠的毛细血管密度明显低于对照组(P<0.01)。(3)免疫蛋白印迹法检测结果表明,Cat S-I组小鼠中,PPAR-γ、p-Akt、p-e NOS和VEGF等蛋白表达水平与对照组比较明显减少(P<0.05)。结论:Cat S-I调控缺血性血管新生可能与PPAR-γ激活及磷脂酰肌醇激酶(PI3K)/Akt/e NOS信号通路有关。
Objectives： To investigate the potential mechanism of cathepsins S inhibitor（Cat S-I） affected ischemia-induced neovascularization in experimental mice. Methods： 8 week-old wild type（C57/BL6） mice were randomly divided into 2 groups： Control group, the mice received basic diet with intraperitoneal injection of 5% carboxymethylcellulose sodium and Cat S-I group, the mice received basic diet with intraperitoneal injection of Cat S-I（1 mg/kg·d）; all animals were treated for 17 days. n=20 in each group. Hind limb ischemia model was established at 3 days after injection in both groups. Blood flow was measured by laser Doppler blood flow analyzer, the ratio of ischemic area to non-ischemic area was calculated; protein expressions of peroxidase proliferation activated receptors-γ（PPAR-γ）, p-Akt, p-e NOS and VEGF were examined by Western blot analysis at day 4 after the operation; frozen section of ischemic skeletal muscle was taken at 7 days after operation to measure capillary density by immunohistochemistry.Results：（1） Cat S-I inhibited blood flow recovery. Compared with Control group, Cat S-I group had slower blood flow recovery in ischemic hind limb, P〈0.05.（2） Cat S-I inhibited capillary formation. At 14 days after operation, capillary formation in non-ischemic skeletal muscles was similar between 2 groups, P 0.05; while in ischemic skeletal muscles, capillary density was lower in Cat S-I group than Control group, P〈0.01.（3） Compared with Control group, Cat S-I group showed decreased protein expressions of PPAR-γ, p-Akt, p-Enos and VEGF, P〈0.05.Conclusions： Cat S-I regulated ischemia-induced neovascularization might be related to PPAR-γ activation and PI3 K/Akt/e NOS signaling pathway in experimental mice.
作者
李洋
朴颖
类延娜
成宪武
李香
LI Yang;PIAO Ying;LEI Yan-na;CHENG Xian-wu;LI Xiang(Department of Cardiology, Yanbian University Affiliated Hospital, Yanji （133000）, Jilin, China)
出处
《中国循环杂志》
CSCD
北大核心
2018年第4期395-399,共5页
Chinese Circulation Journal
基金
国家自然科学基金资助项目(81660240)
