Tian Fengyan, Fu Xi, Gao Jinzhi, Zhang Cai, Ning Qin, Luo Xiaoping
Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2012 Feb;32(1):107-112. doi: 10.1007/s11596-012-0019-5. Epub 2012 Jan 27.
In previous study, glutaric acid (GA) induced apoptosis of primary striatal neuron in vitro. In order to investigate the neurotoxic effects of GA on neonatal rat corpus striatum and the possible mechanism, 34 male pups were randomly assigned to NS group, low dose GA (LGA, 5 μmol GA/g body weight) group and high dose GA (HGA, 10 μmol GA/g body weight) group. These pups were subcutaneously administered with three injections from postnatal day 3 to 22 at 7:30 am, 15:00 pm and 22:30 pm and killed 12 h after the last injection. Microscopic pathology in corpus striatum was evaluated by HE staining. The apoptotic cells were identified by TUNEL staining. The transcript levels of caspase-3, 8, 9, Bax, Bcl-2 were detected by using real-time PCR and the protein levels of procaspase-3 and the active fraction were evaluated by Western blotting. In LGA and HGA groups, ventricle collapse, cortical atrophy by a macroscope and interstitial edema, vacuolations, widened perivascular space of bilateral striatum by a microscope were observed. TUNEL assay revealed that the apoptotic cells were increased in LGA and HGA groups. The transcript of caspase-3 was up-regulated to 2.5 fold, accompanied by the up-regulation of caspase-9, Bax and down-regulation of Bcl-2. The protein levels of procaspase-3 and the active fraction were up-regulated in LGA and HGA groups. The rat model for GA I showed mitochondrial apoptotic pathway may be involved in the GA-induced striatal lesion. Further studies should be taken to investigate the underlying mechanisms.
在先前的研究中，戊二酸（GA）在体外可诱导原代纹状体神经元凋亡。为了研究GA对新生大鼠纹状体的神经毒性作用及其可能的机制，将34只雄性幼崽随机分为生理盐水组（NS组）、低剂量GA组（LGA，5 μmol GA/克体重）和高剂量GA组（HGA，10 μmol GA/克体重）。这些幼崽在出生后第3天至22天，每天上午7:30、下午15:00和晚上22:30进行三次皮下注射，并在最后一次注射后12小时处死。通过苏木精-伊红（HE）染色评估纹状体的微观病理学。通过末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色鉴定凋亡细胞。使用实时聚合酶链反应（PCR）检测半胱天冬酶-3、8、9、Bax、Bcl-2的转录水平，并通过蛋白质免疫印迹法评估前半胱天冬酶-3及其活性片段的蛋白质水平。在LGA组和HGA组中，肉眼可见脑室塌陷、皮质萎缩，显微镜下可见双侧纹状体间质水肿、空泡形成、血管周围间隙增宽。TUNEL检测显示，LGA组和HGA组的凋亡细胞增加。半胱天冬酶-3的转录上调至2.5倍，同时伴有半胱天冬酶-9、Bax的上调和Bcl-2的下调。LGA组和HGA组前半胱天冬酶-3及其活性片段的蛋白质水平上调。GA诱导的纹状体损伤可能涉及线粒体凋亡途径的大鼠模型。应进一步研究其潜在机制。
