DHA调节人脑血管周细胞氧糖剥夺-复氧复糖时血管生成素表达的机制：与SSeCKS的关系  

Mechanism underlying docosahexaenoic acid-induced regulation of angiopoietin expression in human brain vascular pericytes subjected to oxygen-glucose deprivation and restoration： the relationship with SSeCKS

作　　者：余艳丽[1] 夏中元[1] 方海滨[1] 周斌[1] 雷少青[1] 

机构地区：[1]武汉大学人民医院麻醉科,430060

出　　处：《中华麻醉学杂志》2017年第2期188-191,共4页Chinese Journal of Anesthesiology

摘　　要：目的评价二十二碳六烯酸（DHA）调节人脑血管周细胞（HBVPs）氧糖剥夺-复氧复糖时血管生成素（Ang）表达的机制与Src抑制的蛋白激酶C底物（SSeCKS）的关系。方法HBVPs以密度2×105个/ml分别接种于96孔板或6孔板，采用随机数字表法分为5组（n＝18）：正常对照组（C组）、氧糖剥夺-复氧复糖组（OGD/R组）、DHA组（D组）、SSeCKS基因沉默（S组）、SSeCKS基因沉默＋DHA组（SD组）。氧糖剥夺-复氧复糖损伤模型制备方法：采用无糖-无血清培养液，置于94%N2-5%CO2-1%O2条件下缺氧缺糖24 h，再置于高糖DMEM培养基常氧环境中复氧复糖6 h。D组于缺氧前1 h加入DHA，终浓度40 μmol/L。S组和SD组采用siRNA进行SSeCKS基因沉默，随后SD组于缺氧前1 h加入DHA，终浓度40 μmol/L。于复氧6 h时，采用CCK-8法确定细胞存活率，采用ELISA法检测LDH漏出量，采用Western blot法检测SSeCKS、Ang-1和Ang-2的表达。结果与C组比较，OGD/R组细胞存活率降低，LDH漏出量增加，SSeCKS和Ang-1表达下调，Ang-2表达上调，Ang-1/Ang-2比值降低，S组SSeCKS表达下调（P〈0.05）。与OGD/R组比较，D组细胞存活率升高，LDH漏出量降低，SSeCKS和Ang-1表达上调，Ang-2表达下调，Ang-1/Ang-2比值升高（P〈0.05），SD组上述指标差异无统计学意义（P〉0.05）。与D组比较，SD组细胞存活率降低，LDH漏出量升高，SSeCKS和Ang-1表达下调，Ang-2表达上调，Ang-1/Ang-2比值降低（P〈0.05）。结论DHA增加HBVPs氧糖剥夺-复氧复糖时Ang-1/Ang-2比值的机制可能完全与上调SSeCKS表达有关。Objective To evaluate the relationship between the mechanism underlying docosahexaenoic acid （DHA） -induced regulation of angiopoietin expression and Src-suppressed C kinase substrate （SSeCKS） in human brain vascular pericytes （HBVPs） subjected to oxygen-glucose deprivation and resto- ration （OGD/R）. Methods HBVPs were seeded in 96-well or in 6-well plates at a density of 2×10^5 cells/ ml and divided into 5 groups （n= 18 each） using a random number table： control group （group C） , OGD/ R group, DHA group （group D）, SSeCKS gene silencing group （group S） and SSeCKS gene silencing plus DHA group （group SD）. The model of OGD/R injury was established as follows： the cells were subjected to O2-glucose deprivation for 24 h in glucose- and serum-free culture medium aerated with 94% N2- 5%CO2-1%O2 followed by restoration of O2-glucose supply for 6 h in high-glucose DMEM culture medium in normal atmosphere. DHA was added at 1 h before hypoxia with the final concentration of 40 μmol/L in group D. Small interfering RNA induced SSeCKS gene silencing in S and SD groups. Subsequently, DHA with the final concentration of 40 μmol/L was added at 1 h before hypoxia in group SD. At 6 h of reoxygen- ation, the cell survival rate was determined by CCK-8 assay, the amount of LDH released was detected using ELISA, and the expression of SSeCKS, angiopoietin-1 （Ang-1） and Ang-2 was detected by Western blot. Results Compared with group C, the cell survival rate was significantly decreased, the amount of LDH released was increased, the expression of SSeCKS and Ang-1 was down-regulated, the expression of Ang-2 was up-regulated, and Ang-1/Ang-2 ratio was decreased in group OGD/R, and the expression of SSeCKS was down-regulated in group S （P〈0.05）. Compared with group OGD/R, the cell survival rate was significantly increased, the amount of LDH released was decreased, the expression of SSeCKS and Ang-1 was up-regulated, the expression of Ang-2 was down-regulated, and Ang-1/Ang-2 ratio was inc

关 键 词：二十二碳六烯酸类 A型激酶锚定蛋白质类 促血管生成素类 周细胞 脑 再灌注损伤 

分 类 号：R743[医药卫生—神经病学与精神病学]