氟比洛芬酯对局灶性脑缺血再灌注大鼠皮质磷酸化环磷酸腺苷反应元件结合蛋白表达的上调作用

Flurbiprofen up-reguating expression of phosphor-cyclic adenosine monophosphate response ele-ment binding protein during focal cerebral ischemia/reperfusion in rat cerebral cortex

目的观察大鼠局灶性脑缺血再灌注后立即给予氟比洛芬酯（FA）治疗对大脑皮质磷酸化环磷酸腺苷反应元件结合蛋白（p—cREB）表达的影响及对缺血再灌注（I—R）损伤的保护作用。方法雄性sD大鼠45只，随机均分为5组：假手术组（sham组）、缺血再灌注组（I．R组）、脂微球组（Lip组）、FA10mg／，kg组（FAl0组）和FA20m∥蚝组（FA20组）。脑缺血后立即分别静脉注射相应剂量FA及其脂微球载体，制作大脑中动脉栓塞（McA0）模型，再灌注后24、48、72h观察神经功能评分，并于72h后处死大鼠，取大脑皮质用westemblot方法分析瞬时受体电位通道6（TRPc6）和p．cREB的表达；取大脑切片行2％2，3，5一氯化三苯四唑（1TI℃）染色计算脑梗死体积。结果（1）F10和砣0组TRPC6和p—cREB表达均显著高于I—R组（P〈0．01）。（2）再灌注后24、48、72h，F10（12．30±0．84、12．67±1．02、13．17±0．91）和F20组（12．83±1．05、13．00±0．68、13．33±0．71）神经功能评分均显著高于I—R组（6．50±0．76、7．67±0．80、7．83±0．70）（P〈0．01）。（3）I-R组脑梗死体积（24．12±1．98）％显著大于F10组（12．42±1．10）％和F20组（11．73±1．06）％（JP〈O．01）。结论FA可能通过TRPC6／cREB通路明显减轻大鼠局灶性脑I—R损伤。

Objective To explore the effect of flurbiprofen axetil （FA） on the expression of phos- phorylated cyclic AMP response element binding protein （p-CREB） and the protective effects of FA admin- istrated immediately after reperfusion on focal cerebral ischemia-repeffusion （I-R） injury in rats. Methods Forty-five male SD rats were randomly divided into 5 groups ： sham, I-R, lipo-microballoons （Lip） , FA 10 mg/kg and FA 20 mg/kg with 9 rats each. The dose of FA or lipo-microballoons solvent （ the vehicle for FA ） was intravenously administered to the corresponding groups immediately after the establishment of cer- ebral I-R model by middle cerebral artery occlusion （MCAO） for 120 rain. The neurological outcome was evaluated at 24, 48 and 72 h after reperfusion. The brain infarct volume percentage （BIVP） was then as- sessed after 2% TIC staining following the last neurological outcome evaluation. The expression of Transi- ent receptor potential channel 6 （TRPC6） and p-CREB in the ipsilateral cerebral cortex was detected by using Western blotting at 72 h after reperfusion. Results （1） The expression levels of TRPC6 and p-CREB in FA 10 mg/kg and FA 20 mg/kg groups were significantly higher than in I-R group （P 〈 0. 01 ） ; （2） Neuropsychological scores in FA 10 mg/kg group [ （ 12. 30 ＋ 0. 84）, （ 12. 67 ＋ 1.02） and （13.17 ±0.91）] and FA 20 mg/kg group [（12.83±1.05）, （13.00 ±0.68） and （13.33 ±0.71）] were significantly higher than those in I-R group [ （6. 50 ±0. 76）, （7.67 ±0. 80） and （7.83 ±0. 70） ] at 24, 48 and 72 h after reperfusion （ P 〈 0. 01 ） ; （ 3 ） The BIVP in FA 10 mg/kg group （ 12.42 ± 1.10） % and FA 20 mg/kg group （11.73 ± 1.06）% was significantly lower than that in I-R group （24. 12 ± 1.98 ） % at 72 h after reperfusion （ P 〈 0. 01 ）. Conclusion FA treatment may significantly reduce the fo- cal cerebral I-R injury induced by MCAO in rats through TRPC6/CREB pathway.