血塞通对局灶性脑缺血再灌注大鼠VEGF/VEGFR2的调节

Effect of Xuesaitong on VEGF/VEGFR2 pathway in rats with middle cerebral artery occlusion/reperfusion

目的探究血塞通对局灶性脑缺血再灌注(MCAO/R)大鼠血管内皮生长因子(VEGF)/血管内皮细胞生长因子受体2(VEGFR2)通路的影响。方法125只大鼠按随机数字表法分为假手术组,模型组,血塞通低(0.25 mg/mL)、中(0.5 mg/mL)、高(1.0 mg/mL)剂量组,每组25只。检测建模前后,CT灌注成像检测大鼠局部脑血流量、侧支循环(CC)血流代偿程度;建模后,大鼠神经功能,氯化三苯基四氮唑法(TTC)测脑梗死体积,苏木精-伊红染色法(HE)观察大鼠脑组织病理学状态,实时定量PCR法(RT-PCR)、蛋白免疫印迹法检测脑组织中VEGF、VEGFR2表达量。结果与建模1 d前比较,建模8 d后模型组局部脑血流量减少(16.39±1.16)m L/100 g vs.(21.69±2.89)mL/100 g,血塞通中、高剂量组局部脑血流量增加(37.26±2.63)、(43.06±1.69)m L/100 g vs.(22.15±2.93)、(21.98±1.73)、CC血流代偿程度减少(2.17±0.33)、(0.86±0.09)vs.(3.11±0.52)、(3.09±0.47),差异有统计学意义(P<0.05)。建模前1 d,与假手术组比较,模型组、血塞通各剂量组局部脑血流量减少、CC血流代偿程度增加(P<0.05)。建模8 d后,与假手术组比较,模型组、血塞通各剂量组局部脑血流量减少、CC血流代偿程度、神经功能缺损评分、脑梗死体积、VEGF、VEGFR2表达量升高(P<0.05);与模型组比较,血塞通各剂量组局部脑血流量、神经功能缺损评分、脑梗死体积降低、VEGF、VEGFR2表达量增多、CC血流代偿程度减少,呈剂量依赖性(P<0.05)。假手术组脑组织结构清晰,细胞形态正常,模型组细胞凋亡严重,随着血塞通浓度的升高,细胞形态逐渐好转。结论血塞通注射液预处理对MCAO/R大鼠脑侧支循环有保护作用,可能与VEGF/VEGFR2通路蛋白表达有关。

Objective To investigate the effects of Xuesaitong on vascular endothelial growth factor(VEGF)/vascular endothelial growth factor receptor 2(VEGFR2)pathway in rats with middle cerebral artery occlusion/reperfusion(MCAO/R).Methods Rats were randomly divided into sham-operated group,model group,Xuesaitong low(0.25 mg/mL),medium(0.5 mg/mL)and high dose(1.0 mg/mL)groups according to the random number table method;the regional cerebral blood flow and the compensation degree of CC blood flow in rats were examined by CT perfusion imaging before and after modeling.After the model was established,the neurological deficits were assessed in each group and the volume of cerebral infarction was determined by triphenyltetrazolium chloride(TTC)staining;hematoxylin-eosin staining(HE)was used to observe the pathological changes of brain tissue in rats of each group.Real-time quantitative PCR(RT-PCR)and Western blotting were used to detect the expressions of VEGF and VEGFR2 in brain tissues.Results Compared with 1 d before modeling,the regional cerebral blood flow decreased in the modeled 8 d model group(16.39±1.16)mL/100 g vs.(21.69±2.89)mL/100 g(P<0.05),and the regional cerebral blood flew increased(37.26±2.63),(43.06±1.69)mL/100 g vs.(22.15±2.93),(21.98±1.73)mL/100 g and the CC blood flow compensation decreased(2.17±0.33),(0.86±0.09)vs.(3.11±0.52),(3.09±0.47)in the middle and high dose groups of Xuesaitong(P<0.05).1 day before modeling,compared with the sham operation group,the regional cerebral blood flow decreased and the CC blood flow compensation increased in the model group and Xuesaitong group(p<0.05).After 8 days of modeling,compared with the sham operation group,the regional cerebral blood flow reduction,CC blood flow compensation degree,neurological deficit score,cerebral infarction volume,VEGF,VEGFR2 expression increased in the model group and Xuesaitong group(p<0.05);compared with the model group,the regional cerebral blood flow,neurological deficit score,cerebral infarction volume decreased,VEGF,VEGFR2 expression increased,CC blood flow compensation decreased,and the dose was decreased(P<0.05).In the sham operation group,the brain tissue structure was clear,the cell morphology was normal,and the apoptosis of the model group was severe.With the increase of the blood transfusion concentration,the cell morphology gradually improved.Conclusion Xuesaitong injection pretreatment had protective effect on CC in MCAO/R rats,which might be related to the expression of VEGF/VEGFR2 path-way protein.