重组腺病毒-肝细胞生长因子基因治疗延缓自发性高血压大鼠心室重构及对炎症的影响

目的该试验旨在探讨重组腺病毒-肝细胞生长因子(Ad-HGF)基因局部心肌注射对自发性高血压大鼠(SHR)左心室重构及炎症的影响。方法实验共分5组,其中自发性高血压大鼠做为心肌纤维化模型,被随机分成4组,每组6只,(1)阳性对照组(P组);(2)空白腺病毒组(A组null-Ad5,0.1 ml);(3)低剂量组(L组1×109Pfu.ml-1Ad5-HGF,0.1 ml);(4)高剂量组(H组5×109 Pfu.ml-1 Ad5-HGF,0.1 ml);另6只WKY(wista-kyoto)鼠作为正常对照组(N组);A、L,H组SHR开胸,左心室壁5点分别注射0.1 ml null-Ad5、1×109Pfu.ml-1Ad5-HGF 5×109Pfu.ml-1Ad5-HGF,基因转染后继续饲养,4周后处死。测量自发性高血压大鼠左心室质量指数,测定大鼠左室收缩压、舒张末压、压力变化最大上升和下降速率;免疫组化观察CD34(新生血管密度);ELISA法测量血浆IL-1,CRP,TNF-α;Western blot法测定HGF,NF-κB p50亚基。结果 (1)左心室重量指数:与WKY鼠对比,SHR鼠左心室质量指数明显增加,而重组腺病毒-肝细胞生长因子治疗后SHR鼠左心室质量指数明显减少。(2)血流动力学检测:与SHR模型鼠对比,重组腺病毒-肝细胞生长因子治疗组大鼠左室收缩压、压力变化最大上升和下降速率均明显升高,舒张末压明显降低。(3)心肌间质血管密度:与SHR模型鼠对比,重组腺病毒-肝细胞生长因子治疗组大鼠心肌间质血管密度明显增加。(4)炎症因子及NF-κB p50蛋白:重组腺病毒-肝细胞生长因子治疗组与SHR模型鼠相比,血清白介素-1、CRP、肿瘤坏死因子-α明显降低,心肌组织NF-κB p50蛋白表达显著降低。结论重组腺病毒-肝细胞生长因子改善了自发性高血压大鼠的心功能。抑制炎症因子表达并促进间质血管可能是重组腺病毒-肝细胞生长因子改善心室重构的重要机制之一。

Transmyocardial injection adenovirus-mediated human hepatocyte growth factor attenuate left ventricular remodeling in spontaneously hypertensive rats models

Objective To investigate the therapeutic effect on attenuating left vetricular remodeling （LVR） and the optimal dose-effect relationship in spontaneously hypertensive rats （SHR） after being given different doses of adenovirus-mediated human hcpatocyte growth factor （Ad-HGF） transmyocardial injection. Methods Totally twenty-four SHR were randomly divided into four groups： Positive control group（P group） ;Adenovirs group（A group,null-Ad5 ,0.1 ml） ;Low-dose group（L group,1 x 10^9 Pfu · m1-1 Ads-HGF,0.1 ml） ;Highdose group（ H group ,5 x 10^9 Pfu · ml- 1 Ad5 -HGF ,0.1 ml）. Normal control group（ N group） consised of 6 Wista- Kyoto （WKY） rats ; Null-Ad or Ad-HGF was transfected into the left ventricle wall at five different points by direct injection, a left anterolateral thoraeotomy was performed to visualize the LV free wall. Four weeks after surgery,we made immunohistochemical examination of CD34, Western blot analysis of NF-KB p50, and ELLSA analysis of serum IL-1, CRP,TNF-a. Results The LVMI were significantly decresed in the AdHGF group, compared with the Positive control group. Left ventricular end-diastolic pressure （LVEDP） showed a significant decrease in the Ad-HGF group , compared with the Positive control group. The vessal density of myocardial was significantly increase in the Ad-HGF group ,compared with the Positive control group. The serum of IL-1, CRP,TNF-ot and myocardial NF-KB proteins were significantly reduced in the Ad-HGF group ,compared with the Positive control group. Conclusion Our results demonstrate the that gene transfection of B Ad-HGF attenuated the left vetricular remodeling,which has proved that Ad-HGF has angiogenic ,anti-inflammation biological functions.