华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2010年
1期
73-77
,共5页
冯新民%何世银%樊红%谢纪文%沈霖
馮新民%何世銀%樊紅%謝紀文%瀋霖
풍신민%하세은%번홍%사기문%침림
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
聚花過路黃%腦微血管內皮細胞%覈因子κBp65%細胞間黏附分子1%神經保護
취화과로황%뇌미혈관내피세포%핵인자κBp65%세포간점부분자1%신경보호
Lysimachia congestifloa hemsl%brain microvascular endothelial cells%NF-κB p65%ICAM-1%neuroprotection
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 microvascular 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-glucose deprivation,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);NF-κB 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NF-κB 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.
目的 通過測定聚花過路黃(Lysimachia congestifloa hemsl,LCH)對原代大鼠腦微血管內皮細胞(brain microvascular endothelial cells,BMECs)糖-氧剝奪誘導下NF-κB p65蛋白及下遊靶基因ICAM-1錶達的變化,探討其可能的細胞保護機製.方法 選用初生的Wistar大鼠,建立腦微血管內皮細胞培養模型,以糖-氧剝奪方法 (oxygen-glucose deprivation,OGD)複製體外模擬的腦缺血再灌註模型,觀察試驗各組(即正常組、模型組、LCH低劑量組及LCH高劑量組)缺氧缺糖6 h、複氧複糖18 h後BMECs的活性(CCK-8比色法)變化,相差顯微鏡下BMECs的細胞形態學改變,免疫組化測定NF-κB p65蛋白錶達的變化以及熒光定量RT-PCR測定NF-κB p65 mRNA、ICAM-1 mRNA錶達的變化.結果 BMECs活性,LCH高、低劑量組明顯高于模型組(均P＜0.01);BMECs的細胞損傷變化,LCH高劑量組明顯輕于模型組;NF-κB p65暘性錶達,LCH高、低劑量組分彆明顯低于模型組(均P＜0.01);NF-κB p65 mRNA、ICAM-1 mRNA錶達,LCH高、低劑量組明顯低于模型組(均P＜0.01),LCH低劑量組高于LCH高劑量組(P＜0.01). 結論 LCH對糖-氧剝奪誘導下的腦微血管內皮細胞損傷具有保護作用,其機製可能與其抑製NF-κB p65的活化及其下遊靶基因ICAM-1的異常錶達有關.其抑製效應可能存在一定的量-效關繫.
목적 통과측정취화과로황(Lysimachia congestifloa hemsl,LCH)대원대대서뇌미혈관내피세포(brain microvascular endothelial cells,BMECs)당-양박탈유도하NF-κB p65단백급하유파기인ICAM-1표체적변화,탐토기가능적세포보호궤제.방법 선용초생적Wistar대서,건립뇌미혈관내피세포배양모형,이당-양박탈방법 (oxygen-glucose deprivation,OGD)복제체외모의적뇌결혈재관주모형,관찰시험각조(즉정상조、모형조、LCH저제량조급LCH고제량조)결양결당6 h、복양복당18 h후BMECs적활성(CCK-8비색법)변화,상차현미경하BMECs적세포형태학개변,면역조화측정NF-κB p65단백표체적변화이급형광정량RT-PCR측정NF-κB p65 mRNA、ICAM-1 mRNA표체적변화.결과 BMECs활성,LCH고、저제량조명현고우모형조(균P＜0.01);BMECs적세포손상변화,LCH고제량조명현경우모형조;NF-κB p65양성표체,LCH고、저제량조분별명현저우모형조(균P＜0.01);NF-κB p65 mRNA、ICAM-1 mRNA표체,LCH고、저제량조명현저우모형조(균P＜0.01),LCH저제량조고우LCH고제량조(P＜0.01). 결론 LCH대당-양박탈유도하적뇌미혈관내피세포손상구유보호작용,기궤제가능여기억제NF-κB p65적활화급기하유파기인ICAM-1적이상표체유관.기억제효응가능존재일정적량-효관계.
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-dose LCH group or low-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 cytoprotective role in OGD-induced 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.
