miR-103a对癫痫大鼠海马组织星形胶质细胞活化的影响

Effect of miR-103a on the Activation of Astrocytes in Hippocampus of Epilepsy Rats

为了考察miR-103a对癫痫大鼠海马组织星形胶质细胞活化的影响。本研究通过腹腔注射氯化锂和毛果芸香碱诱导癫痫大鼠模型,对大鼠脑室内注射miR-103a抑制剂来敲低miR-103a的表达;采用免疫组织化学染色检测大鼠海马组织中胶质纤维酸性蛋白(GFAP)的阳性表达;采用RT-qPCR和Western blotting方法检测大鼠海马组织中miR-103a、脑源性神经营养因子(BDNF)、GFAP、TNF-α和IL-6的m RNA和蛋白表达;苏木精-伊红(HE)染色评价海马组织病变程度;Nissl染色检测神经元存活情况;TUNEL染色检测神经元的凋亡。结果显示,癫痫大鼠海马组织中miR-103a被上调。下调miR-103a抑制癫痫大鼠海马组织中GFAP的mRNA和蛋白表达,且抑制癫痫大鼠海马神经元的病理损伤,但能促进癫痫大鼠海马神经元的存活并抑制其凋亡。此外,下调miR-103a还抑制癫痫大鼠海马组织中IL-6和TNF-α的表达,并促进癫痫大鼠海马组织中BDNF的表达。本研究表明,靶向沉默miR-103a可以抑制癫痫大鼠海马组织中星形胶质细胞的活化并改善神经元的病理损伤。

To investigate the effect of miR-103 a on the activation of astrocytes in hippocampus of epilepsy rats,in this study,a rat model of epilepsy was induced by intraperitoneal injection of lithium chloride and pilocarpine.Intracerebroventricular injection of miR-103 a inhibitor to knock down miR-103 a expression.Immunohisto chemical staining was used to detect the positive expression of glial fibrillary acidic protein(GFAP)in rat hippocampus.RT-qPCR and Western blotting were used to detect the m RNA and protein expressions of miR-103 a,brain-derived neurotrophic factor(BDNF),GFAP,TNF-αand IL-6 in rat hippocampus.Hematoxylin-eosin(HE)staining was used to evaluate the degree of hippocampal lesions.Nissl staining was used to detect neuron survival.TUNEL staining was used to detect neuronal apoptosis.The results showed that miR-103 a was up-regulated in hippocampus of epileptic rats.Down-regulating miR-103 a not only inhibited mRNA and protein expression of GFAP in hippocampus of epileptic rats,but also inhibited the pathological damage of hippocampal neurons in epileptic rats.However,down-regulating miR-103 a promoted survival and inhibited apoptosis of hippocampal neurons in epileptic rats.In addition,down-regulating miR-103 a inhibited the expression of IL-6 and TNF-αin hippocampus of epileptic rats,and promoted BDNF expression in hippocampus of epileptic rats.This study shows that targeted silencing of miR-103 a can inhibit the activation of astrocytes in the hippocampus of epileptic rats and inhibit the pathological damage of neurons.