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目的:通过研究homer1a基因敲除小鼠脑缺血再灌注损伤及海马区星形胶质细胞活化、数目形态变化,探讨homer1a基因在脑缺血损伤中的作用及机制。方法:取雄性homer1a基因敲除(Knock Out,KO)小鼠及同窝野生型(Wild Type,WT)小鼠各15只,分为基因敲除假手术组(Sham Knock Out,SKO,n=3)、基因敲除型缺血2 h再灌注24 h组(Model Knock Out,MKO,n=12)、野生型假手术组(Sham Wild Type,SWT,n=3)及野生型缺血2 h再灌24h组(Model Wild Type,MWT,n=12)。线栓法闭塞小鼠大脑中动脉制作脑缺血再灌注损伤模型(middle cerebral artery occlusion and reperfusion,MCAO/R),在缺血再灌注损伤前(0 h)及缺血再灌注后3 h、6 h、12 h、24 h后进行改良版神经损伤严重性评分(modified Neurological severity scores,m NSS)、2,3,5—氯化三苯基四氮唑(2,3,5triphenyltetrazolium chloride,TTC)染色、苏木素—伊红染色(Hematoxylin-eosin staining,HE)、原位末端转移酶标记技术(terminal deoxynucleotidyl transferase(Td T)-mediated deoxyuridine triphosphate(d UTP)nick end labeling,TUNEL)检测及免疫荧光染色观察海马区星形胶质细胞神经纤维酸性蛋白(Glial Fibrillary Acidic Protein,GFAP)改变。结果:SKO组、SWT组行为学m NSS评分均为0分,TTC染色未见梗死灶。TUNLE及GFAP染色阳性细胞数很少且未见统计学差异(P>0.05)。脑缺血再灌注24 h后,MKO组m NSS评分较MWT组高;TTC染色MKO组较MWT组梗死百分比高;MKO组较MWT组TUNEL凋亡率高;GFAP免疫荧光染色阳性数MKO组少于MWT组,且均有统计学差异(P<0.05)。结论:homer1a基因敲除加重了小鼠脑缺血再灌注损伤,星形胶质细胞可能参与并发挥复杂作用。
OBJECTIVE: To investigate the role of homer1a gene in cerebral ischemia-reperfusion injury and its mechanism by studying the activation and morphological changes of astrocytes in hippocampus after cerebral ischemia-reperfusion injury in homer1a knockout mice. Methods Fifteen male homer1a knockout (KO) mice and 15 wild type mice (WT) were divided into Sham Knock Out (SKO, n = 3), model knockout ischemia (MKO, n = 12), Sham Wild Type (SWT, n = 3) and wild-type ischemia 2 h reperfusion 24h group (Model Wild Type, MWT, n = 12). The middle cerebral artery occlusion and reperfusion (MCAO / R) model was established by occlusion of the middle cerebral artery (MCAO / R) The modified Neurological severity scores (mNSS), 2,3,5triphenyltetrazolium chloride (TTC) ) Staining, hematoxylin-eosin staining (HE), terminal deoxynucleotidyl transferase (Td T) -mediated deoxyuridine triphosphate (d UTP) nick end labeling (TUNEL) The changes of astrocyte glial fibrillary acidic protein (GFAP) in hippocampus were observed by staining. Results: The scores of behavioral m NSS in SKO group and SWT group were all 0, and no infarction was found in TTC staining. TUNLE and GFAP staining positive cells were few and no significant difference (P> 0.05). MNSS score of MKO group was higher than that of MWT group after 24 h of cerebral ischemia-reperfusion; infarction percentage of MKO group was higher than that of MWT group in MKN group; apoptosis rate of TUNEL in MKO group was higher than MWT group; GFAP immunofluorescence positive MKO group was less In the MWT group, there were significant differences (P <0.05). Conclusion: knockdown of homer1a aggravates cerebral ischemia-reperfusion injury in mice, and astrocytes may play a complex role.