脑泰方对脑缺血再灌注大鼠HIF-1α/VEGF的调节作用

目的从HIF-1α/VEGF通路观察脑泰方对局灶性脑缺血再灌注大鼠的治疗性血管新生样作用。方法将120只SD大鼠随机分为正常组（n=12）、假手术组（n=12）、脑缺血再灌注损伤组（模型组,n=48）和脑泰方组（n=48）。采用线栓法复制大鼠脑缺血再灌注损伤模型,模型组和脑泰方组再分为再灌注时间1、3、5、7天四个亚组（n=12）。通过免疫荧光染色方法观察血管新生的现象,采用RT-PCR方法检测HIF-1α、VEGF-A和VEGFⅡ受体的表达。结果脑泰方组的缺血区有大量血管新生的标记,染色结果为vWF（红）和Brdu（绿）双染;脑缺血再灌注后第1天,与模型组比较,脑泰方治疗组HIF-1α蛋白表达显著增强（P〈0.01）;第3天,VEGF蛋白表达开始增强,脑泰方组与模型组比较,差异有统计学意义（P〈0.01）;第5天,VEGFR蛋白表达升高,与模型组比较,脑泰方组表达最强（P〈0.01）;第7天,各组VEGF和HIF-1α蛋白表达开始下降。结论脑泰方通过提高HIF-1α表达,激活HIF-1α/VEGF信号传导通路,使缺血后血管新生作用加强,以达到治疗性血管新生样作用。

Regulation of Naotai Recipe on the Expression of HIF-loJVEGF Signaling Pathway in CereBral Ischemia/Reperfusion Rats

Objective To observe the therapeutic angiogenesis effect of Naotai Recipe （NR） on local ischemia/reperfusion （I/R） injury of rats by observing signaling pathway of hypoxia-inducible factor- 1α （HIF-1α and vascular endothelial growth factor （VEGF）. Methods Totally 120 Sprague-Dawley （SD） rats were randomly divided into 4 groups, namely, the normal control group （n = 12）, the sham-operation group （n =12）, the I/R model group （n =48）, and the NR group （n =48）. Cerebral I/R injury models were established using thread suture method. Rats in the I/R model group and the NR group were sub-divided into 4 sub-groups according to the 1 st, 3rd, 5th, and 7th I/R day （n = 12）. The phenomenon of neovascu- lization was observed by immunofluorescence staining. The protein and mRNA expression levels of HIF- 1α, VEGF-A, and VEGFR 11 receptor were detected by RT-PCR. Results There were a large amount of labels for neovasculization in the ischemic area of the NR group. Double-immunofluorescence labeling vWF （red） and BrdU （green） ] was observed in the NR group. Compared with the model group, the HIF- 1α protein expression was obviously enhanced on the 1 st day of I/R （P 〈0.01 ）, and the VEGF protein ex- pression started to enhance on the 3rd day in the NR group （P 〈0.01 ）. The VEGFR protein expression level was the highest in the NR group on the 5th day of I/R （P 〈0.01 ）. The protein expression of VEGF and HIF-1α started to decrease on the 7th day of I/R. Conclusion NR could strengthen angiogenesis after I/R by elevating the expression of HIF-1 α and activating HIF-1α/VEGF signaling pathway.