糖尿病对大鼠脑缺血再灌注后缓激肽B1和B2受体表达及作用的影响  

Effects of diabetes on the expression and function of bradykinin 1 and 2 receptors after cerebral ischemia /reperfusion in rats

作　　者：桑红菲 邱忠明[2] 刘玲[1] 许丽丽[1] 张君[1] 张灏[1] 谢怡[1] 张仁良[1] 

机构地区：[1]南京大学医学院附属金陵医院(南京军区南京总医院)神经内科,南京医学硕士研究生210002 [2]第二军医大学金陵医院(南京军区南京总医院)神经内科

出　　处：《医学研究生学报》2015年第5期458-464,共7页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81100870);江苏省自然科学基金(BK2011663)

摘　　要：目的糖尿病基础上发生脑缺血再灌注损伤后,缓激肽B1受体(bradyldnin B1 receptor,B1R)和缓激肽B2受体(bradykinin B2 receptor,B2R)表达及作用的研究尚少。文中旨在观察比较非糖尿病和糖尿病大鼠脑缺血再灌注后B1R和B2R的表达差异及作用变化。方法构建非糖尿病大鼠和2型糖尿病大鼠局灶性脑缺血再灌注(ischemia/reperfusion,I/R)模型,各41只,测试大鼠体重和生化指标。按照随机数字表法分别将8只非糖尿病大鼠及8只2型糖尿病大鼠分为2组:非糖尿病对照组和非糖尿病脑I/R 24 h组,糖尿病对照组和糖尿病脑I/R 24 h组,每组4只。实时荧光定量PCR检测再灌注24 h后B1R和B2R的表达。另将33只非糖尿病大鼠及33只2型糖尿病大鼠以随机数字表法各分为4组:假手术组(n=6)、等渗盐水组(n=9)、B1R拮抗剂组(n=9)和B2R拮抗剂组(n=9),I/R 24 h后,采用NSS评估神经功能缺损程度,TTC染色观察脑梗死体积,TUNEL染色测定细胞凋亡,Fluoro-Jade C染色检测神经元变性。结果 41只糖尿病大鼠造模后3、7、14 d空腹血糖[(23.45±5.01)、(23.71±4.87)、(22.72±4.11)mmol/L]较非糖尿病大鼠[(5.77±0.75)、(6.05±0.69)、(7.15±1.09)mmol/L]明显增加(P<0.05);14 d时血胆固醇[(4.59±3.43)mmol/L]、胰岛素[(67.26±12.02)pmol/L]较非糖尿病大鼠[(1.58±0.37)mmol/L、(25.34±4.88)pmol/L]明显升高(P<0.05),而三酰甘油水平差异无统计学意义(P>0.05)。糖尿病大鼠脑I/R 24 h后脑组织中B1R mRNA水平较非糖尿病大鼠升高更加明显(P<0.01),而B2R mRNA水平则相对下降(P<0.05)。非糖尿病B2R拮抗剂组NSS评分、梗死体积百分比、损伤和凋亡细胞数目较等渗盐水组明显减少(P<0.05);糖尿病B1R拮抗剂组NSS评分、梗死体积百分比、损伤和凋亡细胞数目较等渗盐水组明显减少(P<0.05)。结论非糖尿病大鼠脑I/R损伤诱导了B2R mRNA的显著上调,抑制B2R有效地减少脑梗死体积及减轻组织细胞损伤;糖尿病大鼠的脑I/R损伤诱�Objective There is little research focusing on the expression and function of bradykinin 1 receptor （ B1R ） and bradykinin 2 receptor （ B2R） after cerebral ischemia/reperfusion on the basis of diabetes .The aim of this study was to compare the ex-pression difference and function change of B 1R and B2R in non-dia-betic and diabetic rats . Methods The cerebral ischemia/reperfu-sion model was established on 41 non-diabetic and type 2 diabetic rats, the weight and the biochemical index were measured on these two types of rats .8 non-diabetic rats and 8 diabetic rats were respec- 〈br〉 tively assigned to two groups according to random number tables：control group and I/R 24 h group, 4 in each group.Real-time PCR was performed to observe the expressions of two receptors at 24 h after reperfusion .Then, 33 non-diabetic rats and 33 diabetic rats were randomly divided into 4 groups respectively, including sham group （n=6）, saline group （n=9）, B1R antagonist group （n=9） and B2R antagonist group （n=9）.At 24 hours after cerebral I/R, neurological deficiency was evaluated by neurological severity scores （ NSS）;infarct volume was observed by TTC staining;cell apoptosis was determined by TUNEL staining;neuron degeneration was de-tected by Fluoro-Jade C staining. Results Glucoses of diabetics at 3, 7, 14 d after model establishment [（23.45 ±5.01）, （23.71 ±4.87）, （22.72 ±4.11） mmol/L] were obviously elevated compared with non-diabetics [（5.77 ±0.75）, （6.05 ±0.69）, （7.15 ±1.09） mmol/L];blood cholesterin [（4.59 ±3.43） mmol/L] and insulin [（67.26 ±12.02） pmol/L] at 14 d after model establishment were evidently incresaed in comparison to those in non-diabetics [（1.58 ±0.37） mmol/L, （25.34 ±4.88） pmol/L] （P＆lt;0.05）, while no significant difference was found in the blood triglyceride of diabetics between them （P＆gt;0.05）.Compared with non-diabetics, diabetics suffered from more apparent up-regulation of B1R mRNA （P＆lt;0.01） but relative

关 键 词：2型糖尿病 脑缺血再灌注 缓激肽B1受体 缓激肽B2受体 

分 类 号：R587.1[医药卫生—内分泌]