CAJ | 학술논문

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、24 h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、24 h)iNOS的阳性表达IA值B组(1851.38 ±242.12、2005.41±290.52、2625.28±385.21、3142.50 ±425.72)显著低于A组(1950.25±298.26、2232.45±305.28、325 1.22±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、24 h)B组PAF的血清浓度(15.36±2.12、18.56 ±3.28、28.21±4.26、22.48±4.21) μg/L显著低于A组(20.52±4.26、28.25±6.18、36.08±7.45、30.26±6.02)μg/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.
목적 탐토은행협제취물(GBE)대대서국조성뇌결혈재관주손상적보호작용급기궤제.방법 장80지청길급웅성SD대서수궤분위2조:대조조(A조,급여생리염수치료,n=40지),은행협제취물간예조(B조,급여등량적은행협제취물,n =40지),매조재분4개아조,즉재관주후1、3、6、24 h조,매개아조10지.제작대서우측대뇌중동맥결혈재관주모형.응용소목소-이홍(HE)염색법관찰뇌조직형태학변화급면역조직화학SABC법검측유도형일양화담합매(iNOS)재뇌조직중적표체；응용매련면역흡부시험(ELISA)법검측혈소판활화인자(PAF)재혈청중적농도변화.결과 HE염색:A조발현명현적경사조、신경세포배사、조망,신경세포외형난이변인,대부분세포결구소실.B조형태상대정상,손상교A조경.iNOS면역조직화학결과:결혈재관주1h후가견소량iNOS양성표체세포,3、6、24h가견iNOS양성표체세포명현증다,병차개시출현핵이위현상,포괄신경원、신경효질세포、혈관내피세포등,각시간점(1、3、6、24 h)iNOS적양성표체IA치B조(1851.38 ±242.12、2005.41±290.52、2625.28±385.21、3142.50 ±425.72)현저저우A조(1950.25±298.26、2232.45±305.28、325 1.22±425.26、3965.36±521.62) (P ＜0.05).재관주후PAF적혈청농도승고,6h체고봉,24h개시하강,재관주후각시간점(1、3、6、24 h)B조PAF적혈청농도(15.36±2.12、18.56 ±3.28、28.21±4.26、22.48±4.21) μg/L현저저우A조(20.52±4.26、28.25±6.18、36.08±7.45、30.26±6.02)μg/L(P ＜0.05).결론 iNOS화PAF재대서뇌결혈재관주후균현저증고.은행협제취물가능통과억제PAF화iNOS적표체감경뇌결혈재관주손상,은행협제취물대대서결혈재관주후적뇌조직유현저적보호작용.
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 SpragueDawley 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 subgrouops,namely,1 h,3 h,6 h and 24 h after reperfusion,10 in each sub-group.The model of right middle cerebral artery occlusion (MCAO) and reperfusion was made.Pathological changes in brain morphology were observed by Hematoxylin and Eosin (HE) staining; immunohitochemical method was used to detect 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 vacuolatin were observed,neuronal shape was difficult to identify,and most of the cell structures disappeared 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 cells 1 h after reperfusion,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 ) μg/L than in group A (20.52 ± 4.26,28.25 ± 6.18,36.08 ± 7.45,30.26 ± 6.02) μg/L ( P ＜ 0.05).Conclusion iNOS,and PAF change significantly after the cerebral ischemia-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.