r-OPN通过调控PTEN-AKT通路对大鼠脑出血损伤的作用机制研究

Study on mechanism of r-OPN on cerebral hemorrhage injury in rats by regulating the PTEN-AKT pathway

目的探讨重组骨桥蛋白(r-OPN)通过调控第10号染色体同源丢失性磷酸酶张力蛋白基因(PTEN)/蛋白激酶B(Akt)通路对脑出血(ICH)损伤的作用机制。方法80只SD大鼠随机分为假手术组、模型组、r-OPN组、r-OPN+抑制剂组,每组20只。用自体血注入法建立ICH大鼠模型;r-OPN组和r-OPN+抑制剂组大鼠分别侧脑室注射相应药物,模型组和假手术组大鼠注射等量PBS。于建模后24 h、48 h、72 h给大鼠进行改良神经功能缺损评分(m NSS)。建模后72 h检测大鼠的脑组织含水量,HE染色观察脑组织病理改变,TUNEL染色检测脑组织细胞凋亡率;qRT-PCR和Western blotting法分别检测脑组织相关mRNA和蛋白表达水平。结果与模型组比较,r-OPN组大鼠各时间点m NSS评分、脑组织含水量、细胞凋亡率降低(均P<0.05),脑组织病理改变明显减轻,p-Akt、细胞淋巴瘤/白血病-2基因(Bcl-2)mRNA和蛋白表达水平升高,PTEN、Bcl-2相关X蛋白(Bax)mRNA和蛋白表达水平降低(均P<0.05)。r-OPN+抑制剂可降低部分r-OPN的改善效果。结论r-OPN可能通过调控PTEN/Akt信号通路,减少ICH后神经细胞凋亡,减轻脑组织损伤,改善神经功能。

Objective To investigate the mechanism of recombinant osteopontin(r-OPN)on intracerebral hemorrhage(ICH)injury by regulating homologous loss phosphatase tensin gene(PTEN)/protein kinase B(Akt)pathway on chromosome 10.Methods 80 rats were randomly divided into sham operation group,model group,r-OPN group and r-OPN+inhibitor group.ICH rat model was established by autologous blood injection.The rats in r-OPN group and r-OPN+inhibitor group were injected with corresponding drugs in lateral ventricle respectively,and the rats in model group and sham operation group were injected with the same amount of PBS.The rats were given modified neurological deficit score(m NSS)24 h,48 h and 72 h after modeling.72 hours after modeling,the water content of brain tissue was detected,the pathological changes of brain tissue were observed by HE staining,and the apoptosis rate of brain tissue was detected by TUNEL staining.The expression levels of brain related mRNA and protein were detected by qRT-PCR and Western blotting,respectively.Results Compared with the model group,the m NSS score,brain water content and apoptosis rate of r-OPN group decreased at each time point(all P<0.05),the pathological changes of brain tissue were significantly reduced,the expression levels of p-Akt,bcl-2 gene(Bcl-2)mRNA and protein were increased,and the expression levels of PTEN,bcl-2 related X protein(Bax)mRNA and protein were decreased(all P<0.05).r-OPN+inhibitor could reduce the improvement effect of some r-OPN.Conclusion r-OPN may reduce neuronal apoptosis after ICH,reduce brain injury and improve neurological function by regulating PTEN/Akt signaling pathway.