摘要
目的探讨重组腺相关病毒协同表达双基因:抗菌肽(PR39)和肾上腺髓质素(adrenomedullin,ADM),即r AAVPR39-ADM对大鼠脑缺血/再灌注(ischemia/reperfusion,I/R)损伤的治疗作用。方法离体实验:人脐静脉内皮细胞培养,行血管成型实验。在体实验:建立大鼠脑I/R模型(脑缺血2 h后再灌注)。SD大鼠随机分为:Sham组、生理盐水组(I/R+NS)、空病毒组(I/R+AAV)及治疗组(I/R+r AAV-PR39-ADM组)。I/R组,再灌注24 h后股静脉分别给予等量生理盐水、空病毒及r AAV-PR39-ADM。术后1、2、3、4周,行头颅MRI、神经功能缺损评分、TTC染色及HE染色等方法评价r AAV-PR39-ADM对脑I/R损伤的疗效。结果离体实验中,r AAV-PR39-ADM同对照及空病毒组相比,有明显的促血管成型作用。在体实验中,头颅MRI检查证实模型制备成功。与Sham组相比,各时间点I/R组的神经功能缺损评分及脑组织梗塞的体积均明显增高(P<0.01),I/R组术后即刻神经功能缺损评分差异无统计学意义(P>0.05)。与生理盐水组及空病毒组相比,治疗组各时间点神经功能缺损评分及梗塞体积均明显减小(P<0.01),空病毒组与生理盐水组比较,梗塞面积及神经功能缺损评分差异无统计学意义(P>0.05)。HE染色显示,治疗组的神经元细胞数目与生理盐水组和空病毒组相比明显丰富,缺血区新生血管明显增多。结论 r AAV-PR39-ADM可能通过促进血管新生,改善脑I/R后局部供血及侧枝循环,保护神经元,促进血管再生,减轻I/R脑损伤。
Aim To investigate the curative effect of rAAV-PR39-ADM,which co-expressed the gene of an-tibacterial peptide （PR39 ） and adrenomedullin （ADM）,in a rat cerebral ischemia/reperfusion （I/R） injury.Methods In vitro,Matrigel angiogenesis as-say was made with human umbilical vein endothelial cells.In vivo,the cerebral I/R model was established＆nbsp;by the occlusion of the cerebral artery for 2h and then reperfused for 24 h.SD rats were randomly divided in-to sham group,I/R＋normal saline group,I/R＋null virus （AAV ） group, and IR ＋rAAV-PR39-ADM group.rAAV-PR39-ADM,saline and null virus were administered through the femoral vein after 24 h of the reperfusion in I/R group.MRI,neurological deficit＆nbsp;score,TTC and HE staining were measured respective-ly 1 ,2,3 and 4 weeks after the injection in order to e-valuate the therapeutic efficacy.Results In vitro, rAAV-PR39-ADM group had significant angiogenic effect compared with sham group and null virus group. In vivo,successful I/R model was verified by the ima-ges of MRI.Compared with sham group,the nerve function defect score and the cerebral infarction size in each time nodes were significantly raised in I/R groups （P〈0.01）.There was no significant difference in in-farct size and nerve function defect score between I/R＋normal saline group and I/R ＋null virus （AAV ）＆nbsp;group,and obviously,the IR ＋rAAV-PR39-ADM group lowered these indexes compared with the other two groups.HE staining showed that the number of neurons,new capillaries vessels of I/R ＋PR39-ADM group were significantly more than those in group I/R and group I/R ＋null virus.Conclusion The treat-ment of rAAV-PR39-ADM promotes vascular forma-tion,neuron protection and reduces the infarct size in the model of cerebral ischemia/reperfusion.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2015年第5期641-647,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81170185)
