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Cerebral lymphatic blockage aggravates apoptosis of cortical neurons after subarachnoid hemorrhage in rats in vivo

WANG Xuan, GAO Xiang-Dong, GAO Bing, JIA Li-Li, YANG Ming-Feng, ZHANG Yan-Bo, SUN Bao- Liang

Key Laboratory of Cerebral Microcirculation in Universities of Shandong (Taishan Medical College), Department of Neurology, Affiliated Hospital of Taishan Medical College, Taian 271000, China； Department of Neurology, Zhangqiu People’s Hospital, Zhangqiu 250200, China

Abstract

This present study was performed to investigate the influence of cerebral lymphatic blockage (CLB) on apoptosis of corticalneurons after subarachnoid hemorrhage (SAH) in rats in vivo. Healthy adult Wistar rats were randomly divided into normal controlgroup, SAH group and SAH + CLB group. SAH model was made by double injection of autologous blood into the cisterna magna. Onthe third day after the second cisternal injection, morphological changes of cortical cells were observed by hematoxylin-eosin (HE)combined with propidium iodide (PI) staining. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)method was applied to determine in situ apoptosis in the cerebral cortex. Immunohistochemistry was conducted to detect the expressionof Caspase-3 and Bcl-2 in cortical neurons. HE and PI staining showed that cortical neurons of SAH rats were partly shrinkable;the nuclei showed wavy, folded or wrinkled appearance, and some nuclei had the shape of crescent. The cortical neurons in SAH + CLBgroup distributed sparsely and the nuclear fragmentation, apoptotic bodies were found, surrounded by the formation of vacuoles. Thenumbers of TUNEL-positive cells in SAH group and SAH + CLB group were higher than that in the normal control group, while thenumber in SAH + CLB group was significantly higher than that in the SAH group. Caspase-3 expressions in SAH group and SAH +CLB group were higher than that in the normal control group, while the expression in SAH + CLB group was significantly higher thanthat in the SAH group. Bcl-2 expressions in SAH group and SAH + CLB group were higher than that in the normal control group, whilethe expression in the SAH + CLB group was significantly lower than that in SAH group. The results obtained suggest that CLBexacerbates the apoptosis of cortical neurons in rats after SAH by up-regulating Caspase-3 expression and down-regulating Bcl-2expression.

Key words: cerebral lymphatic drainage; subarachnoid hemorrhage; apoptosis; Caspase-3; Bcl-2

Received: 2009-11-19　　Accepted: 2010-02-01

Corresponding author: 王轩　　E-mail: blsun88@163.com

Citing This Article：

WANG Xuan, GAO Xiang-Dong, GAO Bing, JIA Li-Li, YANG Ming-Feng, ZHANG Yan-Bo, SUN Bao- Liang. Cerebral lymphatic blockage aggravates apoptosis of cortical neurons after subarachnoid hemorrhage in rats in vivo. Acta Physiol Sin 2010; 62 (2): 109-114 (in Chinese with English abstract).