Feng Shu-Qing, Zong Shao-Yun, Liu Jia-Xin, Chen Yang, Xu Rong, Yin Xin, Zhao Rong, Li Ying, Luo Ting-Ting
Department of Ultrasound, The First People's Hospital of Yunnan Province.
Department of Ultrasound, The Affiliated Hospital of Kunming University of Science and Technology.
Biol Pharm Bull. 2019 May 1;42(5):692-702. doi: 10.1248/bpb.b18-00628. Epub 2019 Mar 2.
Endoplasmic reticulum (ER) stress-mediated apoptosis pathway is considered to play a vital role in mediating stroke and other cerebrovascular diseases. Previous studies have showed that vascular endothelial growth factor (VEGF) antagonism reduced cerebral ischemic-reperfusion (CI/R) damage, but whether attenuation of ER stress-induced apoptosis is contributing to its mechanisms remains elusive. Our study aimed to investigate the protective effect of VEGF antagonism on CI/R-induced injury. First, oxygen-glucose deprivation and re-oxygenation (OGD/R) BEND3 cell model was constructed to estimate small interfering RNA (siRNA)-VEGF on damage of endothelial cells. Next, in animal model, CI/R mice were induced by middle cerebral artery occlusion (MCAO) for 2 h followed by 24 h reperfusion to investigate cerebral tissue damage. For treatment group, mice received 100 µg/kg anti-VEGF antibodies at 30 min before MCAO, followed by 24 h reperfusion. Our findings demonstrated that pre-administration of siRNA-VEGF before OGD/R changed the biological characteristics of BEND3 cells, reversed the levels of X-box binding protein-1 (XBP-1) and glucose-regulated protein 78 (GRP78), showing siRNA-VEGF attenuated, at least in part, the oxidative damage in OGD/R cell by down-regulating ER stress. In mice experiment, pre-administration of anti-VEGF antibody reduced the brain infarct volume and edema extent and improved neurological scores outcome of CI/R injury mice. Pathological and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining results also confirmed this protective effect. The expressions of VEGF, CATT/EBP homologous protein (CHOP), inositol requiring enzyme 1α (IRE-1α), and cleaved-caspase12 and c-jun N-terminal kinase (JNK) phosphorylation were also prominently decreased. These results suggested that inhibition of endogenous VEGF attenuates CI/R-induced injury via inhibiting ER stress-mediated apoptosis.
内质网(ER)应激介导的凋亡途径被认为在介导中风和其他脑血管疾病中起着至关重要的作用。先前的研究表明,血管内皮生长因子(VEGF)拮抗作用可减少脑缺血再灌注(CI/R)损伤,但ER应激诱导的凋亡减弱是否是其作用机制仍不清楚。我们的研究旨在探讨VEGF拮抗作用对CI/R诱导损伤的保护作用。首先,构建氧糖剥夺和复氧(OGD/R)BEND3细胞模型,以评估小干扰RNA(siRNA)-VEGF对内皮细胞损伤的影响。接下来,在动物模型中,通过大脑中动脉闭塞(MCAO)2小时,随后再灌注24小时诱导CI/R小鼠,以研究脑组织损伤。对于治疗组,小鼠在MCAO前30分钟接受100μg/kg抗VEGF抗体,随后再灌注24小时。我们的研究结果表明,在OGD/R之前预先给予siRNA-VEGF改变了BEND3细胞的生物学特性,逆转了X盒结合蛋白-1(XBP-1)和葡萄糖调节蛋白78(GRP78)的水平,表明siRNA-VEGF至少部分地通过下调ER应激减轻了OGD/R细胞中的氧化损伤。在小鼠实验中,预先给予抗VEGF抗体可减少脑梗死体积和水肿程度,并改善CI/R损伤小鼠的神经评分结果。病理和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记(TUNEL)染色结果也证实了这种保护作用。VEGF、CATT/EBP同源蛋白(CHOP)、肌醇需求酶1α(IRE-1α)、裂解的半胱天冬酶12和c-jun氨基末端激酶(JNK)磷酸化的表达也显著降低。这些结果表明,抑制内源性VEGF通过抑制ER应激介导的凋亡减轻CI/R诱导的损伤。