摘要
目的探讨吡格列酮（pioglitazone，PIO）对Wistar大鼠大脑皮质神经元氧-葡萄糖剥夺／复氧（oxyge-glucose deprivation／reoxygenation，OGD／R）后过氧化物酶体增殖物激活受体1（peroxisome proliferator—activated receptors，PPAR-y）、核因子KB（nuclearfactor—KB，NF—KB）亚基c—Rel以及抗凋亡因子Bcl—xL表达的影响。方法大鼠大脑皮质神经元体外原代培养，建立OGD／R模型，分为正常组、OGD4h／R6h组、OGD4h／R12h、OGD4h／R6h＋PIO组、OGD4h／R6h＋PPARγ抑制剂T0070907（TIO）组、OGD4h／R12h＋PIO组以及OGD4h／R12h＋TIO组。采用蛋白质印迹法检测神经元内PPARγ／和NF—KBc—Rel蛋白表达，采用逆转录一聚合酶链反应检测神经元内Bcl—xLmRNA表达。结果蛋白质印迹分析显示，OGD／R后PPAR~／和NF—KBc—Rel蛋白表达水平较正常组显著性增高（P〈0．01），给予PIO后PPAR3＇和NF—KBc—Rel蛋白表达水平进一步增加，显著高于单纯OGD4h／R6h组和OGD4h／R12h组（P〈0．01），而给予TIO后PPARγ和NF—KBc—Rel蛋白表达减少，显著性低于单纯OGD／R组（P〈0．05）以及相应PIO组（P〈0．01）。逆转录一聚合酶链反应分析显示，OGD4h／R6h组Bcl—xLmRNA表达水平较正常组显著性增高（P〈0．01），PIO组Bcl．xLmRNA表达水平显著性高于单纯OGD4h／R6h组和OGD4h／R12h组（P〈0．01），给予TIO后Bcl—xLmRNA表达减少，显著性低于单纯OGD／R组（P〈0．05）以及相应PIO组（P〈0．01）。结论PIO可上调培养大脑皮质神经元OGD／R后PPAR／蛋白表达，进而增加NF—KB亚基c—Rel调控的抗凋亡因子Bcl—xLmRNA表达，这可能是PPAR＇y神经保护作用的部分机制。
Objective To investigate the effect of pioglitazone （PIO） on the expression of peroxisome proliferators-activated receptor （PPAR/）, nuclear factor-KB （NF-Kt3） c-Re1 and anti-apoptosis factor Bcl-xL in cultured Wistar rat cortical neurons after oxygen-glucose deprivation/reoxygenation （OGD/R）. Methods The rat cerebral cortical neurons were primarily cultured in vitro and a model of OGD/R was induced. The rats were divided into 7 groups： normal, OGD 4 h/R 6 h, OGD 4 h/R 12 h, OGD 4 h/P, 6 h ＋ PIO, OGD 4 h/P, 6 h ＋ PPAR~/ inhibitor T0070907 （TIO）, OGD 4 h/R 12 h ＋ PIO, and OGD 4 h/R 12 h ＋ TIO groups. Western blot was used to detect the expression of PPAR-y and NF-KB c-Rel protein in neurons. Reverse transcription-polymerase chain reaction assay was used to detect the expression of Bcl-xL mRNA in neurons. Results Western blot analysis showed that the expression levels of PPAR/ and NF-KB c-Rel proteins after OGD/R were increased significantly compared to those of the normal group （P 〈 0. 01 ）. After giving PIO, the expression levels of PPAR/ and NF-KB c-Rel proteins were further increased, and they were significantly higher than those in the OGD 4 h/R 6 h and OGD 4 h/R 12 h groups （P 〈 0. 01）, and after giving TIO, the expression levels of PPAR＇,/ and NF-KB c-Rel proteins were decreased, and they were significantly lower than those in the OGD/R group （P 〈 0. 05） and the corresponding PIO group （P 〈 0. 01）. Reverse transcription-polymerase chain reaction analysis showed that the expression level of Bcl-xL mRNA in the OGD 4 h/R 6 h group was significantly higher than that in the normal group （P 〈0. 01）. The expression level of Bcl-xL mRNA in the P10 group was significantly higher than that in the OGD 4 h/R 6 h and OGD 4 h/R 12 h groups （P 〈0. 01）. After giving TIO, the expression of Bcl-xL mRNA was decreased. It was significantly lower than that in the OGD/R and corresponding PIO groups （P 〈 0. 05 and P 〈 0. 01 ）. Conclusions PIO can upregulate
出处
《国际脑血管病杂志》
北大核心
2013年第5期357-362,共6页
International Journal of Cerebrovascular Diseases
基金
重庆市卫生局资助项目（渝卫科教[2003]61号,03-2.179）
