SCAD重组腺病毒对自发性高血压大鼠血管重构的影响

目的探究尾静脉注射短链酰基辅酶A脱氢酶(SCAD)重组腺病毒能否改善自发性高血压大鼠(SHR)血管重构。方法实验分为6组:Wistar+NS组、Wistar+GFP组、Wistar+Ad-SCAD组、SHR+NS组、SHR+GFP组和SHR+Ad-SCAD组。腺病毒包装的SCAD和GFP纯化后,以尾静脉方式注射给药8周。采用超声心动图检测大鼠心功能情况;采用无创血压仪检测大鼠的血压变化;主动脉HE染色、天狼星红染色、DHE染色、TUNEL染色、EVG染色,观察血管重构。采用实时荧光定量PCR检测相关基因mRNA表达变化、Western blot检测相关蛋白表达变化,观察游离脂肪酸、一氧化氮(NO)和ATP含量变化。结果 (1)尾静脉注射SCAD重组腺病毒,大鼠主动脉中SCAD蛋白出现过表达,mRNA水平增高,SCAD酶活性增加;(2)在病理状态下,增加主动脉中SCAD表达,可以降低血压,改善心功能,改善血管管腔大小,减少胶原沉积,减少血管活性氧(ROS)的生成和细胞凋亡;(3)在病理状态下,SCAD的表达增加可减少血清和主动脉中游离脂肪酸的含量,增加组织中的ATP水平,激活内皮型一氧化氮合酶(eNOS)磷酸化,增加主动脉NO的生成。结论自发性高血压大鼠主动脉SCAD的表达增加能逆转高血压血管重构,可能与其减少血清中游离脂肪酸的含量、增加NO水平、减少活性氧生成、消除氧化应激有关。

Effect of short chain acly-CoA dehydrogenase recombinant adenovirus on vascular remodeling in spontaneously hypertensive rats

Aim To investigate whether the recombinant adenovirus with short chain acyl CoA dehydrogenase (SCAD) injected via tail vein can improve the vascular remodeling in spontaneously hypertensive rats (SHR). Methods The experiment was divided into 6 groups:Wistar+NS group, Wistar+GFP group, Wistar+Ad-SCAD group, SHR+NS group, SHR+GFP group and SHR+Ad-SCAD group. After purification of SCAD and GFP packed with adenovirus, the drug was injected by tail vein for 8 weeks. The cardiac function of rats was detected by echocardiography. The blood pressure changes of rats were detected by non-invasive blood pressure meter. HE staining, Sirius red staining, DHE staining, TUNEL staining, EVG staining, were used to observe the phenomenon of vascular remodeling. The expression of related proteins was detected by Western blot, and mRNA expression was detected by RT-PCR. The free fatty acid (FFA), nitric oxide (NO) content and ATP content were observed. Results (1) After SCAD recombinant adenovirus was injected via tail vein, the expression of SCAD protein was significantly upregulated in the aorta of rats, meanwhile, the mRNA level was observably increased, and the activity of SCAD was markedly increased. (2) In the pathological state of rats, the rising of SCAD can lower blood pressure, improve heart function and vascular lumen size, reduce collagen deposition, result a poor production of vascular ROS, consequently, give lower to apoptosis. (3) Under pathological conditions, overexpression of SCAD can reduce FFA content of serum and aorta, increase the ATP level in aorta, activate eNOS phosphorylation, increase NO production in aorta. Conclusion The upregulation of SCAD in aorta of SHR can reverse hypertensive vascular remodeling, which may be related to decreasing FFA content of serum, increasing NO levels, reducing ROS production and eliminating oxidative stress.