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谷氨酰胺联合氟比洛芬酯对大鼠局灶性脑缺血再灌注损伤的影响
PROTECTIVE EFFECTS OF COMBINATION OF GLUTAMINE AND FLURBIPROFEN AXETIL ON FOCAL CEREBRAL ISCHEMIA REPERFUSION INJURY IN RATS

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DOI:
作者:
辛志祥，姜虹，朱也森
XIN Zhi-xiang, JIANG Hong，ZHU Ye-sen
作者单位:
上海第九人民医院麻醉科
Department of Anesthesiology，Shanghai Ninth Peoplel’s Hospital Affiliated of Shanghai Jiaotong University,School of Medicine
关键词:
谷氨酰胺；氟比洛芬；脑缺血；再灌注损伤
Glutamine； flurbiprofen； cerebral ischemia； reperfusion injury
摘要:
【摘要】 目的 探讨谷氨酰胺联合氟比洛芬酯对大鼠局灶性脑缺血再灌注损伤的保护作用及机制。 方法 改良线栓法制作大鼠大脑中动脉闭塞（MCAO）2h/再灌注24h模型。将60只雄性SD大鼠随机分为5组(每组12只)。假手术组（Sham组）：不插入线栓，其余操作与缺血再灌注组相同。缺血再灌注组（IR组）:于插入线栓前1h和15min静脉分别注射生理盐水1ml作为对照。谷氨酰胺组（Gln组）: 于插入线栓前1h静脉注射0.75g/kg谷氨酰胺1ml，插入线栓前15min静脉注射生理盐水1ml。氟比洛芬酯组（FPA组）：于插入线栓前1h静脉注射生理盐水1ml，插入线栓前15min静脉注射10mg/kg氟比洛芬酯1ml。谷氨酰胺联合氟比洛芬酯组（Gln+FPA组）：于插入线栓前1h静脉注射0.75g/kg谷氨酰胺1ml,插入线栓前15min静脉注射10mg/kg氟比洛芬酯1ml。MCAO 2h/再灌注24h后TTC染色计算脑梗死率；HE染色观察脑组织学形态学变化；免疫组化法检测脑组织HSP70，NF-КB蛋白。 结果 ①与IR组相比，Gln组、FPA组能缩小MCAO 2h/再灌注24h后脑梗死率（P＜0.05），Gln+FPA组能显著缩小MCAO 2h/再灌注24h后脑梗死率(P<0．01)。②与IR组相比，3个治疗组脑组织损伤均有所减轻，Gln+FPA组减轻程度最显著。③与Sham组比，IR组海马、缺血半暗带HSP70阳性细胞表达明显增加(P<0.01)；与IR组相比，Gln组和Gln+FPA组均显著增加MCAO 2h/再灌注24h后大鼠大脑缺血半暗带及海马组织HSP70的表达(P<0.01)，与IR组相比，FPA组HSP70无明显增加(P>0.05)。④与Sham组相比，IR组缺血半暗带NF-κB蛋白表达明显增加(P<0．01)；与IR组相比，3个治疗组大鼠缺血半暗带中NF-κB蛋白表达均有明显减少(P<0．05或0.01)；与Gln组和FPA组相比，Gln+FPA组大鼠缺血半暗带中NF-κBp65蛋白表达进一步减少(P<0．05)。结论 谷氨酰胺和氟比洛芬酯预处理均能不同程度减轻大鼠局灶性脑缺血再灌注损伤，两者联合应用具有增强作用。
【Abstract】 Objective To investigate the protective effects and mechanisms of combination of glutamine and flurbiprofen axetil(FPA) on focal cerebral ischemia reperfusion injury (CIRI) in adult rats. Methods The modified middle cerebral artery occlusion(MCAO)2h／reperfusion 24h were used in adult rats. 60 SD rats were randomly divided into 5 groups（n=12）.Sham group(sham operation), IR group (normal saline+local cerebral I／R),Gln group (glutamine 0.75g/kg +local cerebral I／R)，FPA group（FPA 10mg/kg +local cerebral I／R）,Gln+FPA group（glutamine 0.75g/kg+FPA 10mg/kg+ local cerebral I／R）. After reperfusion 24h,the histological changes were observed through HE staining. The infarction ratios of the brain were measured by TTC staining. The expression of HSP70 and NF-κB protein were measured by immunohistochemical technique. Reults Compared with IR group，both the brain infarction ratios and the histological damage were significantly decreased in three treated groups，and it was slightest in Gln+FPA group. The expression of HSP70 proteins in hippocampus or IP area of IR group were significantly increased compared with Sham group (P<0.01) .Compared with IR group，Gln and Gln+FPA group could up-regulate the expression of HSP70 proteins(P<0.01)，however，there was no significant difference between IR group and F group(P>0.05). The expression of NF-κB proteins in IP area of IR group were significantly increased compared with Sham group(P<0．01). Compared with IR group，the expression of NF-κB in three treated groups were significantly decreased(P< O．05 or 0.01).Conclusion Pretreatment with glutamine or FPA could protect the neurons from focal CIRI in rats ,and combination of both has a fortified action.

 

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