Graduate School, Anhui University of Chinese Medicine, Hefei 230012, China.
Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui University of Chinese Medicine, Hefei 230012, China.
J Tradit Chin Med. 2023 Oct;43(6):1140-1149. doi: 10.19852/j.cnki.jtcm.20221230.001.
To observe regulatory effect of Naoluoxintong formula (, NLXT) and its split prescriptions on vascular regeneration of rats suffering from cerebral ischemia-reperfusion (IR) syndrome of Qi deficiency with blood stasis (QDBS).
NLXT is the representative prescription of Yiqi Huoxue Tongluo decoction, and NLXT is divided into Yiqi herbs and Huoxue Tongluo herbs according to their efficacies. One hundred and eight specific-pathogen-free, clean-grade, Sprague-Dawley male rats were selected to prepare the classical rat model with QDBS due to middle artery ischemia-reperfusion using the multi-factor compound simulation approach. The animals were classified into sham operation (S), model (M), Nimodping (NMDP), NLXT, YQ and HXTL groups, each having 18 rats. Cerebral ischemia was reperfused after 2 h, and 24 h later, they were administered traditional Chinese medicine treatment for 14 d twice a day. Angiogenesis changes after NLXT administration to middle cerebral artery occlusion-reperfusion (MCAO/R) rats with QDBS were analyzed using the neurological deficit score and hematoxylin-eosin staining. Cerebral infarct area by 2,3,5-Triphenyltetrazolium chloride was detected, and the ultrastructure of the blood vessel in the ischemic frontoparietal cortex was observed by transmission electron microscopy. Angiopoietin 1 (Ang1), angiopoietin 2 (Ang2), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2), platelet endothelial cell adhesion molecule-1 (CD31), angiopoietin receptor 2 (Tie2), and P38 mitogen-activated protein kinase (MAPK) protein levels in the frontal and parietal cortex were quantified by immunofluorescence, reverse transcription-polymerase chain reaction, and Western blotting assays.
Relative to the S group, VEGFA and VEGFR2 levels in the frontal and parietal cortex of group M were increased, and Ang1, Ang2, Tie2, CD31, and p38 MAPK levels remarkably increased ( < 0.05); cerebral infarct area was significant and pathological morphology and ultrastructure damage was obvious. Relative to the group M, VEGFA, VEGFR2, CD31, Ang1, Ang2, and Tie2 expression of group NLXT and NMDP remarkably elevated ( < 0.05) and infarct focus, pathological morphology and ultrastructure were significantly improved; VEGFA and VEGFR2 levels in the groups YQ and HXTL increased, and Ang1, Ang2, CD31, and Tie2 levels remarkably increased ( < 0.05); p38 MAPK levels in the three treatment groups decreased ( < 0.05). Relative to the group NLXT, the expression levels of p38 MAPK in group YQ and group HXTL were significantly increased, and the expression levels of other indicators were significantly decreased ( < 0.05).
NLXT can promote the angiogenesis of the rat model of MCAO/R with QDBS by activating VEGFA and inhibiting P38 MAPK, and the effect is better than that of split prescription groups.
观察脑络欣通方及其拆方对气虚血瘀型脑缺血再灌注（IR）大鼠血管再生的调控作用。
脑络欣通方是益气活血通络方的代表方剂，根据功效将其拆分为益气药和活血通络药。采用多因素复合模拟法制备气虚血瘀型脑缺血再灌注经典大鼠模型，选取 108 只特定病原体-清洁级 Sprague-Dawley 雄性大鼠，采用大脑中动脉缺血再灌注，造模后 2 h 再灌注，再灌注 24 h 后，每天两次给予中药治疗 14 d。采用神经功能缺损评分和苏木精-伊红染色分析脑络欣通方对大脑中动脉闭塞再灌注（MCAO/R）大鼠气虚血瘀证的血管生成变化。用 2,3,5-三苯基氯化四氮唑检测脑梗死面积，用透射电镜观察缺血额顶叶皮质血管的超微结构。采用免疫荧光、逆转录-聚合酶链反应和 Western blot 检测额叶和顶叶皮质中血管生成素 1（Ang1）、血管生成素 2（Ang2）、血管内皮生长因子 A（VEGFA）、血管内皮生长因子受体 2（VEGFR2）、血小板内皮细胞黏附分子-1（CD31）、血管生成素受体 2（Tie2）和 P38 丝裂原活化蛋白激酶（MAPK）的蛋白水平。
与 S 组相比，M 组额叶和顶叶皮质的 VEGFA 和 VEGFR2 水平升高，Ang1、Ang2、Tie2、CD31 和 p38 MAPK 水平显著升高（ < 0.05）；脑梗死面积显著增加，病理形态和超微结构损伤明显。与 M 组相比，NLXT 组和 NMDP 组的 VEGFA、VEGFR2、CD31、Ang1、Ang2 和 Tie2 表达显著升高（ < 0.05），梗死灶、病理形态和超微结构明显改善；YQ 组和 HXTL 组的 VEGFA 和 VEGFR2 水平升高，Ang1、Ang2、CD31 和 Tie2 水平显著升高（ < 0.05）；三组治疗组的 p38 MAPK 水平降低（ < 0.05）。与 NLXT 组相比，YQ 组和 HXTL 组的 p38 MAPK 表达水平显著升高，其他指标表达水平显著降低（ < 0.05）。
脑络欣通方通过激活 VEGFA 和抑制 P38 MAPK 促进 MCAO/R 大鼠模型的血管生成，其作用优于拆方组。
