聚花过路黄对原代大鼠脑微血管内皮细胞糖-氧剥夺诱导下NFκ-B p65蛋白及下游靶基因ICAM-1表达的影响  

Impact of Lysimachia Congestifloa Hemsl on Expressions of NF-κB p65 Protein and Its Downstream Target Gene ICAM-1 mRNA Induced by Oxygen-Glucose Deprivation in Rat Primary Brain Microvascular Endothelial Cells

作　　者：冯新民[1] 何世银[1] 樊红[1] 谢纪文 沈霖[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院1中西医结合科2急诊内科,武汉430022

出　　处：《华中科技大学学报（医学版）》2010年第1期73-77,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省卫生厅科研基金资助项目(No.9325)

摘　　要：目的通过测定聚花过路黄(Lysimachia congestifloa hemsl,LCH)对原代大鼠脑微血管内皮细胞(brain mi-crovascular endothelial cells,BMECs)糖-氧剥夺诱导下NF-κB p65蛋白及下游靶基因ICAM-1表达的变化,探讨其可能的细胞保护机制。方法选用初生的Wistar大鼠,建立脑微血管内皮细胞培养模型,以糖-氧剥夺方法(oxygen-glucosedeprivation,OGD)复制体外模拟的脑缺血再灌注模型,观察试验各组(即正常组、模型组、LCH低剂量组及LCH高剂量组)缺氧缺糖6 h、复氧复糖18 h后BMECs的活性(CCK-8比色法)变化,相差显微镜下BMECs的细胞形态学改变,免疫组化测定NF-κB p65蛋白表达的变化以及荧光定量RT-PCR测定NF-κB p65 mRNA、ICAM-1 mRNA表达的变化。结果BMECs活性,LCH高、低剂量组明显高于模型组(均P<0.01);BMECs的细胞损伤变化,LCH高剂量组明显轻于模型组;NF-κB p65阳性表达,LCH高、低剂量组分别明显低于模型组(均P<0.01);NFκ-B p65 mRNA、ICAM-1 mRNA表达,LCH高、低剂量组明显低于模型组(均P<0.01),LCH低剂量组高于LCH高剂量组(P<0.01)。结论LCH对糖-氧剥夺诱导下的脑微血管内皮细胞损伤具有保护作用,其机制可能与其抑制NFκ-B p65的活化及其下游靶基因ICAM-1的异常表达有关。其抑制效应可能存在一定的量-效关系。Objective To investigate the effects of Lysimachia congestifloa hemsl（LCH）on the expression of NF-κB p65 protein and its downstream target gene ICAM-1 mRNA after oxygen-glucose deprivation（OGD）in primary brain microvascular endothelial cells（BMECs）of rats. Methods Primary BMECs in rats were subjected to OGD in the absence or presence of LCH. The viability of BMECs was assessed by CCK 8,the morphological changes of BMECs were examined under a phase-contrast microscope, the expression level of NF-κB p65 protein in BMECs was detected by using immunohistochemical staining, and the expression level of ICAM 1 mRNA in BMECs was assayed by fluorescent quantitation RT PCR. Results The viability of BMECs in high dose LCH group or low dose LCH group was significantly higher than that in model group（both P〈0.01）. The damage of BMECs in high-dose LCH group was significantly milder than that in model group. The positive expression rate of NF-κB p65 in BMECs of high-close LCH group or tow-dose LCH group was significantly lower than that of model group（both P〈0.01）. The positive expression rate of ICAM 1 mRNA in BMECs of high-dose LCH group was significantly lower than that in model group（P〈0.01） ,and low-dose LCH group（P〈0.01）. Conclusion LCH may play a eytoprotective role in OGD-in duced cell damage by inhibiting the abnormal expression of NF-κB p65 and its downstream target gene ICAM-1 ,and the inhibitory effects may be dose-dependent.

关 键 词：聚花过路黄 脑微血管内皮细胞 核因子ΚBP65 细胞间黏附分子1 神经保护 

分 类 号：R845.2[医药卫生—航空、航天与航海医学]