MiR-153在TPM治疗颞叶癫痫大鼠海马组织中的表达及意义

MiR-153 in TPM treatment of temporal lobe epilepsy rat hippocampal tissue of the expression and significance

目的探讨miRNAs参与颞叶癫痫(TLE)的发生发展,TPM治疗颞叶癫痫中有miR-153的参与及其与凋亡之间的联系。方法利用锂-匹罗卡品诱发癫痫,随机将80只大鼠分为四组:正常对照组,TLE组,TPM低剂量组(40mg/kg)和TPM高剂量组(80 mg/kg)。通过microRNA芯片分析测定上述四组大鼠海马中miRNAs表达谱的改变,HE染色观察各组大鼠海马组织与细胞的形态学改变,并利用免疫蛋白印迹、免疫组化方法测定上述各组海马中caspase-3蛋白的表达水平。结果与对照组相比,TLE大鼠海马中检测到了6个上调的miRNAs(miR-204,-214,-210,-466c,-298,-207)和12个下调的miRNAs(miR-153,-34a,-362,-377,-101a,-382,-598,-488,-383,-376b-3p,-380,-9)。经TPM治疗后,上调的6个miRNAs发生了下调,12个下调的miRNAs则出现了上调,其中miR-153在上调的miRNAs中变化最为显著。此外,在TLE大鼠海马中能够检测到caspase-3的表达水平明显增加,经TPM治疗后其表达水平下降,且TPM高剂量组中大鼠海马caspase-3的表达较TPM低剂量组更低。结论 MiRNAs参与到了TLE的发生发展中,且miR-153有望成为TLE治疗中的重要靶点。MiR-153参与到了癫痫的发生,TPM治疗能够使miR-153发生上调。TPM能够减少大鼠海马中凋亡的发生。

Objective This study aimed to investigate the involvement of miRNAs in the development of TLE,the association of miR-153 and caspase-3with TPM treatment.Methods Li-pilocarpine was used for induction of epilepsy and 80 rats were divided into four groups randomly：1）control,2）TLE group,3）low dose TPM group（40mg/kg）,and 4）high dose TPM group（80mg/kg）.MiRNAs profiling in hippocampus of four groups was detected using gene chip microarray analysis,the morphological changes of hippocampus were observed using H＆E staining,and the expression level of caspase-3protein in hippocampus was detected by western blot and immunohistochemistry（IHC）.Results A total of 6 miRNAs（miR-204,-214,-210,-466 c,-298,-207）were up-regulated and 12 miRNAs（miR-153,-34 a,-362,-377,-101 a,-382,-598,-488,-383,-376b-3p,-380,-9）were down-regulated in TLE rat models compared with control rats.Importantly,these 6up-regulated miRNAs were down-regulated,and the 12down-regulated miRNAs were up-regulated in both low dose and high dose TPM-treated groups.Caspase-3was significantly up-regulated in the TLE group,but downregulated in TPM-treated groups,especially in the high dose group.Conclusion MiRNAs is involved in the development of TLE and miR-153 could be a new therapeutic target in TPM treatment.miR-153 is involved in the development of TLE and TPM treatment could up-regulatemiR-153.TPM could reduce the apoptosis in hippocampus of TLE rats.