Stereotaxic microinjection of adenovirus-mediated human tissue Kallikrein gene reduces apoptosis in a rat model of middle cerebral artery occlusion  

作　　者：Ruiyan Lu Lianhong Yang Qingyu Shen Mei Li Xiangpen Li Ying Peng 

机构地区：[1]Department of Neurology, Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China

出　　处：《Neural Regeneration Research》2008年第8期847-852,共6页中国神经再生研究（英文版）

基　　金：the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, No. 002006006

摘　　要：BACKGROUND：Several studies have demonstrated that Kallikrein gene transfer provides neuroprotection. Whether the neuroprotective effects of human tissue Kallikrein （HTK） are associated with apoptosis remains unclear. OBJECTIVE： To investigate the effects of HTK on apoptosis in the peripheral cerebral infarct region. DESIGN, TIME AND SETTING： The completely randomized grouping, gene engineered, controlled experiment was performed at the Lin Baixin Laboratory Center, the Second Affiliated Hospital of Sun Yat-sun University between September 2007 and April 2008. MATERIALS： Ninety clean, healthy, male, Sprague Dawley rats were included. pUC19-HTK plasmid was constructed in the Laboratory for Neurology, the Second Affiliated Hospital of Sun Yat-sen University, China. bcl-2, bax, caspase-3, and β-actin were designed and purified by Shanghai Shuiyuan Company, China. METHODS： Middle cerebral artery occlusion （MCAO） model was established in all rats. At 72 hours after MCAO model establishment, rats were randomly divided into 3 groups, with 30 rats per group： blank control, saline, and pAdCMV-HTK. The saline and pAdCMV-HTK groups were respectively stereotactically micro-injected with 5 μL physiological saline or pAdCMV-HTK at the area surrounding the cerebral infarction region. Only puncture was performed, without any injection, in the blank control group. MAIN OUTCOME MEASURES： At 72 hours after MCAO establishment, as well as at 24 hours, 72 hours, and 7 days subsequent to treatment, exogenous HTK expression was detected by immunohistochemistry. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）, while mRNA levels of bcl-2, bax, and caspase-3 were detected by reverse transcription-polymerase chain reaction. In addition, neurological severity scores were evaluated prior to and after treatments. RESULTS： Ninety rats were included in the final analysis. HTK was primarily detected in the cytoplasm at 24 hours after pAdCMV-HTK injection. ThereafBACKGROUND：Several studies have demonstrated that Kallikrein gene transfer provides neuroprotection. Whether the neuroprotective effects of human tissue Kallikrein （HTK） are associated with apoptosis remains unclear. OBJECTIVE： To investigate the effects of HTK on apoptosis in the peripheral cerebral infarct region. DESIGN, TIME AND SETTING： The completely randomized grouping, gene engineered, controlled experiment was performed at the Lin Baixin Laboratory Center, the Second Affiliated Hospital of Sun Yat-sun University between September 2007 and April 2008. MATERIALS： Ninety clean, healthy, male, Sprague Dawley rats were included. pUC19-HTK plasmid was constructed in the Laboratory for Neurology, the Second Affiliated Hospital of Sun Yat-sen University, China. bcl-2, bax, caspase-3, and β-actin were designed and purified by Shanghai Shuiyuan Company, China. METHODS： Middle cerebral artery occlusion （MCAO） model was established in all rats. At 72 hours after MCAO model establishment, rats were randomly divided into 3 groups, with 30 rats per group： blank control, saline, and pAdCMV-HTK. The saline and pAdCMV-HTK groups were respectively stereotactically micro-injected with 5 μL physiological saline or pAdCMV-HTK at the area surrounding the cerebral infarction region. Only puncture was performed, without any injection, in the blank control group. MAIN OUTCOME MEASURES： At 72 hours after MCAO establishment, as well as at 24 hours, 72 hours, and 7 days subsequent to treatment, exogenous HTK expression was detected by immunohistochemistry. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）, while mRNA levels of bcl-2, bax, and caspase-3 were detected by reverse transcription-polymerase chain reaction. In addition, neurological severity scores were evaluated prior to and after treatments. RESULTS： Ninety rats were included in the final analysis. HTK was primarily detected in the cytoplasm at 24 hours after pAdCMV-HTK injection. Thereaf

关 键 词：adenovirus-mediated APOPTOSIS apoptotic factor cerebral infarction KALLIKREIN 

分 类 号：R743[医药卫生—神经病学与精神病学]