Zheng Yong-Qiu, Yao Ming-Jiang, Liu Jian-Xun, Song Wen-Ting, Li Lei, Liu Sheng-Bo, Hu Yan, Si Jing-Xi
Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China.
Zhongguo Zhong Yao Za Zhi. 2013 Feb;38(4):585-90.
To observe the effect and mechanism of Huatuo Zaizao extractum (HTZZ) on focal ischemia/reperfusion (I/R) blood-brain barrier injury induced by middle cerebral artery occlusion.
Sixty healthy male adult Sprague-Dawley rats was randomly divided into the sham operation group, the MCAO model group, the Tanakan (20 mg x kg(-1)) group, and high, middle and low-dose HTZZ groups (5, 2.5, 1.25 g x kg(-1)), with 10 in each group and single-dose duodenal administration. Middle cerebral artery occlusion was adopted to establish the rat focal I/R model. After ischemia for 90 min and reperfusion for 24 h, the pathological injury at the ischemia side was observed by HE staining. The blood-brain barrier structure was observed under transmission electron microscope. Expressions of G protein-coupled receptor kinases 2 (GRK2), matrix metalloproteinases 2 (MMP-2) and MMP-9 were detected by western blotting technique.
After 90 min MCAO/24 h reperfusion, penumbra cerebral cortical micro-vessels showed edema, mitochondrial injury, vacuolation, membrane injury and reduction. Along with the changes, sub-cells of G protein-coupled receptor kinase 2 (GRK2) in cortical penumbra brain tissues transferred from cytoplasm to membrane, with increase in expressions of MMP-2 and MMP-9. HTZZ could effectively recover cerebral micro-vascular endothelial edemaand blood-brain barrier ultrastructure injury induced by I/R, reduce expression of functional (membrane coupling) GRK2, and inhibit expressions of MMP-2 and MMP-9.
Cell membrane coupling GRK2 may be the effective target of Huatuo Zaizao extractum.
观察华佗再造丸提取物（HTZZ）对大脑中动脉闭塞所致局灶性缺血/再灌注（I/R）血脑屏障损伤的作用及机制。
将60只健康成年雄性Sprague-Dawley大鼠随机分为假手术组、大脑中动脉闭塞（MCAO）模型组、灯盏细辛组（20 mg·kg⁻¹）和HTZZ高、中、低剂量组（5、2.5、1.25 g·kg⁻¹），每组10只，单剂量十二指肠给药。采用大脑中动脉闭塞法建立大鼠局灶性I/R模型。缺血90分钟再灌注24小时后，通过苏木精-伊红（HE）染色观察缺血侧的病理损伤。在透射电子显微镜下观察血脑屏障结构。采用蛋白质免疫印迹技术检测G蛋白偶联受体激酶2（GRK2）、基质金属蛋白酶2（MMP-2）和MMP-9的表达。
MCAO 90分钟/再灌注24小时后，半暗带大脑皮质微血管出现水肿、线粒体损伤、空泡化、膜损伤和减少。随着这些变化，皮质半暗带脑组织中G蛋白偶联受体激酶2（GRK2）的亚细胞从细胞质转移到细胞膜，MMP-2和MMP-9的表达增加。HTZZ可有效恢复I/R诱导的脑微血管内皮水肿和血脑屏障超微结构损伤，降低功能性（膜偶联）GRK2的表达，并抑制MMP-2和MMP-9的表达。
细胞膜偶联GRK2可能是华佗再造丸提取物的有效靶点。
