摘要
目的探讨银杏叶提取物（GBE）对大鼠局灶性脑缺血再灌注损伤的保护作用及其机制。方法将80只清洁级雄性sD大鼠随机分为2组：对照组（A组，给予生理盐水治疗，n=40只），银杏叶提取物干预组（B组，给予等量的银杏叶提取物，n=40只），每组再分4个亚组，即再灌注后1、3、6、24h组，每个亚组10只。制作大鼠右侧大脑中动脉缺血再灌注模型。应用苏木素-伊红（HE）染色法观察脑组织形态学变化及免疫组织化学SABC法检测诱导型一氧化氮合酶（iNOS）在脑组织中的表达；应用酶联免疫吸附试验（ELISA）法检测血小板活化因子（PAF）在血清中的浓度变化。结果HE染色：A组发现明显的梗死灶、神经细胞坏死、凋亡，神经细胞外形难以辨认，大部分细胞结构消失。B组形态相对正常，损伤较A组轻。iNOS免疫组织化学结果：缺血再灌注1h后可见少量iNOS阳性表达细胞，3、6、24h可见iNOS阳性表达细胞明显增多，并且开始出现核移位现象，包括神经元、神经胶质细胞、血管内皮细胞等，各时间点（1、3、6、24h）iNOS的阳性表达M值B组（1851．38±242．12、2005．41±290．52、2625．28±385．21、3142．504-425．72）显著低于A组（1950．25±298．26、2232．45±305．28、3251．22±425．26、3965．36±521．62）（P〈0．05）。再灌注后PAF的血清浓度升高，6h达高峰，24h开始下降，再灌注后各时间点（1、3、6、24h）B组PAF的血清浓度（15．36±2．12、18．56-4-3．28、28．214-4．26、22．48±4．21）¨g／L显著低于A组（20．52±4．26、28．25±6．18、36．08±7．45、30．26±6．02）斗g／L（P〈0．05）。结论iNOS和PAF在大鼠脑缺血再灌注后均显著增高。银杏叶提取物可能通过抑制PAF和iNOS的表达减轻脑缺血再灌注损伤，银杏叶提取物对大鼠缺血再灌注后的脑组织有显著的保护作用。
Objective To explore the protective effect and mechanism of ginkgo biloba extract （GBE） on brain tissue after local cerebral ischemia and reperfusion in rats. Methods Eight male Sprague- Dawley rats were randomly divided into two groups ： control group （ group A, normal saline injected, n = 40）, GBE intervention group （group B, GBE injected, n = 40）, which were then divided into 4 sub- grouops, namely, 1 h, 3 h, 6 h and 24 h after reperfusion, 10 in each sub-group. The model of right mid- die cerebral artery occlusion （MCAO） and reperfusion was made. Pathological changes in brain morpholo- gy were observed by Hematoxylin and Eosin （HE） staining; immunohitochemical method was used to de- tect the inducible nitric oxide synthase （iNOS） changes; platelet activating factor （PAF） was detected by enzyme linked immunosorbent assay （ELISA）. Results Nerve cell necrosis, apoptosis and cytoplasmic vaeuolatin were observed, neuronal shape was difficult to identify, and most of the cell structures disap- peared in group A. The injury was relieved at the same time points in group B compared with that in group A. Immunohitochemical results showed that there were a few iNOS expression positive ceils 1 h after reper- fusion, which increased at 3 h after referfusion and had nucler translocation. Analysis by Image-pro plus 5. 0 showed that iNOS expression levels in group B （1851.38 ＋ 242. 12, 2005.41 ~ 290.52, 2625.28 ~385.21, 3142. 50 ~ 425.72） were significantly lower than those in group A （ 1950.25 ＋ 298. 26, 2232. 45 ＋305.28, 3251.22 ＋425.26, 3965.36 ~521.62） （P 〈0. 05）, respectively. ELISA results showed that serum PAF levels were increased after ischemia-reperfusion, peaked at 6 h, and decreased at 24 h. Serum PAF level were significantly lower in group B （ 15. 36 ＋ 2. 12, 18. 56 ~ 3.28, 28. 21 ~ 4. 26, 22.48 ~ 4. 21 ） ixg,/L than in group A （20. 52 ~ 4. 26, 28. 25 ~ 6. 18, 36.08 ~ 7.45, 30. 26 ~ 6. 02） ~g/L （P 〈 0. 05）. Conclusion iNOS, and PAF change significantly after the cerebral isehemia-reperfusion in rats, which may be involved in cerebral ischemia-reperfusion injury mechanism. GBE can significantly alleviate the cerebral injury by regulating the expression of iNOS and PAF, suggesting GBE can effectively protect the ischemic cerebral tissues.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第2期259-261,共3页
Chinese Journal of Experimental Surgery
基金
中国博士后基金资助项目（57068）
关键词
银杏叶提取物
脑缺血
再灌注损伤
诱导型一氧化氮合酶
Ginkgo biloba extract
Cerebral ischemia
Reperfusion injury
Inducible nitricoxide synthase
