姜黄素对缺氧再灌注时体外培养星形胶质细胞载脂蛋白E及类视黄醇X受体、肝X受体表达的影响

Effects of curcumin on apoe、rxr and lxr in primary cultured astrocytes underoxygen-glucose deprivation / reoxygenation

目的:本研究旨在观察在姜黄素(Curcumin)对离体缺血再灌注模型中,原代培养新生SD大鼠的大脑皮层星形胶质细胞载脂蛋白E(Apolipoprotein E,Apo E)、类视黄醇X受体(Retinoic X receptor,RXR)和肝X受体(Liver X receptor,LXR)的影响。方法:选取新生SD大鼠,对其大脑皮层的星形胶质细胞进行原代培养,用糖氧剥夺/再灌注(oxygen-glucose deprivation/reoxygenation,OGD/Reoxygenation)方法来构建离体的脑缺血再灌注模型。运用免疫荧光法观察神经胶质原纤维酸性蛋白(GFAP)的表达来鉴定星形胶质细胞。本实验将培养的细胞分为7组:正常对照组;模型组;姜黄素剂量组:2.5umol/L,5umol/L,10umol/L,20umol/L;尼莫地平阳性对照组:0.5umol/L。采用四甲基偶氮唑蓝(MTT)和乳酸脱氢酶(lactate dehydrogenase,LDH)两个指标分别对细胞的活力以及细胞的毒性进行评定。用ELISA法对原代细胞条件培养液中Apo E和RXR、LXR的含量进行测定,并用蛋白质印迹法(Western Blot)对星形胶质细胞Apo E和RXR、LXR的表达水平进行测定。结果:姜黄素可使细胞活力明显增高,LDH释放量显著降低,并使APOE及LXR、RXR的表达上调。其中20umol/L效果最为明显,而最低剂量2.5umol/L几乎为无效剂量。结论:姜黄素对糖氧剥夺再灌注条件下离体星形胶质细胞具有保护作用,此保护作用可能是通过上调APOE及LXR、RXR的表达所引起的。

Objective：This study was designed to investigate the effects of Curcumin on ApoE.RXR and LXR in Primary Cultured Astrocytes Under Oxygen-glucose deprivation/Reoxygenation. Methods ： Primary cultured astrocytes, using the way of oxygen-glucose deprivation/reoxygenation make Cerebral ischemia-reperfusion model in vitro. Through the expression of glial fibriUary acidic protein （GFAP） by using immunofluores- cence to identify astrocytes. In this study, the cultured cells were divided into seven groups： normal control group; the model group; curcu-rain group：2.5 umol / L, 5 umol / L, 10 umol / L, 20 umol / L; nimodipine group： 0.5umol / L. Viability and cytotoxieity of cells to be assessed by MTT and LDH. ELISA method was used to measure the content of ApoE, RXR, LXR in btoth and Western blotting was used to detect the expression of ApoE, RXR, LXR. Results：Curcumin can significantly increased cell viability and reduced LDH release. The ex- pression of ApoE, RXR, LXR was increased. In it, 20 umol / L was most obvious and the lowest dose 2.5 umol / L was almost ineffective. Conlusion：Curcumin has a protective effect for Primary Cultured Astrocytes Under Oxygen-glucose Deprivation/Reoxygenation, This protec- tive effect may be regulated by the expression of APOE LXR, RXR arising.