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目的 筛选和鉴定反复高 GZ暴露大鼠脑损伤相关基因。 方法 16只 SD大鼠随机分成对照组、+GZ重复暴露后 30 min、6 h和 2 4h4组 ,每组 4只。对照组大鼠 G值为 +1GZ,实验组大鼠在动物离心机上经历了 3次 +14GZ/ 45 s(两次间间隔 30 m in)作用。分别于暴露后 30 m in、6 h和2 4h处死大鼠取脑 ,提取总 RNA。取对照组和 +GZ暴露后 6 h组大鼠脑总 RNA,用 m RNA差异显示( DDPCR)的方法 ,筛选 +GZ暴露大鼠脑差异表达基因。用 3组锚定引物和 6组随机引物作不同组合进行 PCR扩增。用 Slot blot方法分析差异表达基因在对照组和 +GZ暴露后 30 m in、6 h和 2 4h组大鼠脑中表达情况。 结果 获得的差异表达片段 ,经斑点杂交进一步筛选 ,克隆了 3个强阳性片段 ,并进行序列分析 ,与 Genebank等数据库进行同源比较 ,没有同源基因。筛选的 3个差异表达基因在+GZ暴露后 6 h和 2 4h组大鼠脑中呈高表达。 结论 用 DDPCR筛选到 3个可能与反复高 GZ暴露大鼠脑损伤相关基因 ,它们在脑损伤的病理过程中可能起一定作用。
Objective To screen and identify the genes related to brain injury induced by repeated high GZ exposure in rats. Methods Twenty-six SD rats were randomly divided into control group, GZ group, 30 rats in each group, 6 h rats and 24 h animals in each group. The G value of the control group was + 1GZ, and the rats in the experimental group experienced three times + 14GZ / 45s on the animal centrifuge (twice between intervals of 30 mins). Rats were sacrificed at 30 min, 6 h and 24 h after exposure respectively, and total RNA was extracted. Total RNA was extracted from rat brain at 6 h after exposure to + GZ, and the differentially expressed genes in brain of + GZ-exposed rats were screened by m RNA differential display (DDPCR). Three groups of anchor primers and six random primers were used for PCR amplification. The expression of differentially expressed genes in rat brain at 30 min, 6 h and 24 h after exposure to + GZ was analyzed by Slot blot. Results The differentially expressed fragments were obtained and further screened by dot blot hybridization. Three strong positive clones were cloned and sequenced. The homology comparison with Genebank database showed that there were no homologous genes. The three differentially expressed genes were highly expressed in rat brain at 6 h and 24 h after + GZ exposure. Conclusion Three genes related to brain injury induced by repeated high GZ exposure in rats were screened by DDPCR, which may play roles in the pathological process of brain injury.