CAJ | 학술논문

目的探讨脂联素(APN)在糖尿病大鼠脑缺血再灌注损伤后血清及脑组织中的表达变化及其作用. 方法 104只SD大鼠按随机数字表法分成正常血糖组(8只)、糖尿病组(8只)、单纯脑缺血再灌注损伤组(I/R组,44只)、糖尿病合并脑缺血再灌注损伤组(DM+I/R组,44只).采用给予糖尿病饮食方法制备糖尿病模型,采用线栓法制备大脑中动脉阻塞(MCAO)缺血再灌注损伤模型.I/R组和DM+I/R组大鼠采用Zea Longa法进行神经行为学评分,并各取4只于脑缺血再灌注损伤前及不同再灌注时间点(3 h、6h、24 h、48 h、72 h、7 d)连续取血测定血清APN浓度变化;剩余40只大鼠再按不同再灌注时间点分为再灌注3h、6h、24 h、72 h、7d5个亚组(每个亚组8只),HE染色观察大鼠脑组织形态学变化,免疫组化染色及Western blotting法检测大鼠缺血脑组织APN蛋白表达. 结果 (1)DM+I/R组大鼠神经功能受损严重,其神经功能学评分[(2.79±0.41)分]明显较I/R组[(1.27±0.45)分]高,差异有统计学意义(P＜0.05).(2)与I/R组相比,DM+I/R组大鼠血清APN水平在再灌注3h、6h、24 h、48 h、72 h、7d时均明显减少,差异均有统计学意义(P＜0.05).(3)DM+I/R组大鼠HE染色示脑组织损伤程度较I/R组明显严重,包括细胞变性、坏死明显.(4)免疫组化染色显示:I/R组和DM+I/R组大鼠缺血脑组织APN表达在再灌注3h时明显增加达高峰,6h时逐渐降低,24h时再次增高并在7d时维持在较高的水平;DM+I/R组大鼠在相同再灌注时间点APN表达均较I/R组明显减少,差异有统计学意义(P＜0.05).Western blotting法检测到的APN蛋白含量变化与免疫组化染色结果相同. 结论 APN在糖尿病大鼠脑缺血再灌注损伤后表达降低,其可能是糖尿病加重脑缺血再灌注损伤的内源性机制之一.
목적 탐토지련소(APN)재당뇨병대서뇌결혈재관주손상후혈청급뇌조직중적표체변화급기작용. 방법 104지SD대서안수궤수자표법분성정상혈당조(8지)、당뇨병조(8지)、단순뇌결혈재관주손상조(I/R조,44지)、당뇨병합병뇌결혈재관주손상조(DM+I/R조,44지).채용급여당뇨병음식방법제비당뇨병모형,채용선전법제비대뇌중동맥조새(MCAO)결혈재관주손상모형.I/R조화DM+I/R조대서채용Zea Longa법진행신경행위학평분,병각취4지우뇌결혈재관주손상전급불동재관주시간점(3 h、6h、24 h、48 h、72 h、7 d)련속취혈측정혈청APN농도변화;잉여40지대서재안불동재관주시간점분위재관주3h、6h、24 h、72 h、7d5개아조(매개아조8지),HE염색관찰대서뇌조직형태학변화,면역조화염색급Western blotting법검측대서결혈뇌조직APN단백표체. 결과 (1)DM+I/R조대서신경공능수손엄중,기신경공능학평분[(2.79±0.41)분]명현교I/R조[(1.27±0.45)분]고,차이유통계학의의(P＜0.05).(2)여I/R조상비,DM+I/R조대서혈청APN수평재재관주3h、6h、24 h、48 h、72 h、7d시균명현감소,차이균유통계학의의(P＜0.05).(3)DM+I/R조대서HE염색시뇌조직손상정도교I/R조명현엄중,포괄세포변성、배사명현.(4)면역조화염색현시:I/R조화DM+I/R조대서결혈뇌조직APN표체재재관주3h시명현증가체고봉,6h시축점강저,24h시재차증고병재7d시유지재교고적수평;DM+I/R조대서재상동재관주시간점APN표체균교I/R조명현감소,차이유통계학의의(P＜0.05).Western blotting법검측도적APN단백함량변화여면역조화염색결과상동. 결론 APN재당뇨병대서뇌결혈재관주손상후표체강저,기가능시당뇨병가중뇌결혈재관주손상적내원성궤제지일.
Objective To investigate the expression of adiponectin (APN) and the clinical significance in type 2 diabetic rats with cerebral ischemic reperfusion damage.Methods One hundred and four SD rats were randomly divided into two groups,including normal diet group (n=52) and diabetic diet group (n=52); middle cerebral artery occlusion was performed in some of the rats to induce ischemia reperfusion injury models.Normal diet group was then randomly divided into euglycemia group (n=8) and ischemia reperfusion group (I/R,n-44); diabetic diet group was randomly divided into diabetes groups (n=8) and diabetes complicated with ischemia reperfusion group (DM+I/R,n=44).The blood was collected to detect the APN changes from the tail vein of rats from diabetes group (n=4) and DM+I/R group (n=4) before MCAO surgery and at 3,6,24,48 and 72 h,and 7 d after reperfusion.The rest 40 rats were randomly divided into subgroups subjected to 90 min of focal ischemia followed by 3,6,24 and 72 h,and 7 d reperfusion (n=8); the neuroethology assessment was determined by Zea Longa method; morphology of brain tissue was observed by HE staining.APN expression in infarction cores was detected by immunohistochemistry and Western blotting.Results The assessment scores of DM+I/R group (2.79±0.41) were significantly higher than those of I/R group (1.27±0.45,P＜0.05).As compared with that in the I/R group,the serum APN level in the DM+I/R group was significantly decreased 3,6,24,48 and 72 h,and 7 d after reperfusion (P＜0.05).Cerebral tissue damage (cellular degeneration and necrosis) in DM+I/R group was more serious as compared with I/R group at the same reperfusion time.In the ischemic hemisphere,APN expression increased at 3 h,decreased at 6 h,and increased again till 24 h after reperfusion,and then it remained at high level up to 7 days after reperfusion.The expression of APN in DM+I/R group was significantly lower than that in the I/R group (P＜0.05).These findings were consistent with the results of Western blotting.Conclusion APN expression decreases after focal cerebral ischemia-reperfusion injury,indicating that APN plays an important role in aggravating ischemic reperfusion injury by diabetes.