LOC389023在难治性颞叶癫痫中分子海绵机制的预测

Forecasting of Molecular Sponge Mechanism Mediated by LOC389023 in Patients with Intractable Temporal Lobe Epilepsy

【目的】研究可能与LOC389023和二肽基肽酶10(DPP10)作用的micro RNA在难治性颞叶癫痫患者皮质中的表达变化,预测LOC389023在颞叶癫痫中的分子海绵机制。【方法】15例脑外伤患者切除的皮质为正常组,26例难治性颞叶癫痫患者行癫痫灶切除的皮质为癫痫组。运用western blot、免疫组织化学染色检测DPP10和电压依赖性钾通道4.3(Kv4.3)的表达变化,采用免疫荧光染色对DPP10和kv4.3进行定位,采用免疫共沉淀分析DPP10与Kv4.3的相关性。运用miRanda、Pita、Target Scan和miRDB软件分别预测与LOC389023以及DPP10相互作用的micro RNA。采用q PCR技术检测LOC389023以及micro RNA的表达变化。【结果】免疫组织化学染色和western blot分析结果显示,DPP10在癫痫组中高表达(P<0.05),Kv4.3则低表达(P<0.05);DPP10与Kv4.3共表达于神经元的细胞膜且两者有相互作用;通过软件预测得到5个候选micro RNA(miR-32-5p,miR-140-5p,miR-367-3p,miR-25-3p和miR-4325);q PCR检测结果显示:在癫痫组中,LOC389023和miR-140-5p表达水平均低于对照组(P<0.05),miR-25-3p和miR-367-3p表达水平均高于对照组(P<0.05),而miR-32-5p和miR-4325无明显变化(P>0.05)。【结论】LOC389023可能以分子海绵的机制调控miR-25-3p和miR-367-3p,继而调控DPP10的表达参与癫痫的形成。

【Objective】To forecast the sponge mechanism mediated by LOC389023 in patients with intractable temporal lobe epilepsy（TLE）,through investigating the expression of microRNA interacted with dipeptidyl peptidase 10（DPP10）and LOC389023.【Methods】The expression of DPP10 and Kv4.3 were detected in 15 temporal neocortex from patients with brain trauma（control group）and in 26 temporal neocortex from patients with intractable TLE（epilepsy group）by western blot（WB）and immunohistochemical（IHC）staining. The location of DPP10 and voltage dependent potassium channel 4.3（Kv4.3）was detected by immunofluorescent（IF）staining. The interaction between DPP10 and Kv4.3 was testified by co-immunoprecipitation（Co-IP）. The expression of microRNA obtained by softwares（miRanda,Pita,Target Scan and miRDB）was detected by q PCR.【Results】IHC and WB showed an increased expression of DPP10（P〈0.05）and a decreased expression of Kv4.3（P〈0.05）in the epilepsy group. IF showed that the DPP10 and the Kv4.3 co-expressed in the membrane and the cytoplasm of neurons. Co-IP showed obvious interaction between the DPP10 and the Kv4.3.Five microRNA（miR-32-5p,miR-140-5p,miR-367-3p,miR-25-3p,miR-4325）were obtained by softwares. No significant differences in the expression of miR-32-5p and miR-4325 were found between epilepsy group and control group by q PCR（P〉0.05）. But decreased expression of LOC389023 and miR-140-5p and increased expression of miR-25-3p and miR-367-3p were found in epilepsygroup compared to control group（P〈0.05）.【Conclusion】miR-25-3p and miR-367-3p may be regulated by LOC389023 through the sponge mechanism followed by altered expression of DPP10 in intractable temporal lobe epilepsy.