摘要
目的丁苯酞氯化钠注射液后处理对X连锁凋亡抑制蛋白(X-inhibitor of apoptosis,XIAP)、Bcl-2/腺病毒E1B19k Da相互作用蛋白3(Bcl-2/adenovirus E1B19k Da interacting protein 3,BNIP3)的作用研究甚少。文中旨在观察丁苯酞氯化钠注射液后处理对局灶性脑缺血再灌注损伤(ischemia reperfusion,IR)大鼠缺血侧海马CA1区XIAP、BNIP3表达的影响,探讨丁苯酞氯化钠注射液的脑保护机制。方法将清洁级雄性SD大鼠65只按随机数字表法分为假手术组(n=13)、IR组(n=13)、丁苯酞组(n=39),丁苯酞组按照剂量不同分为低剂量、中剂量和高剂量亚组,每组13只。IR组:采用改良线栓法制作大鼠大脑中动脉IR模型(缺血2 h,再灌注24 h)。低、中、高剂量亚组在大鼠缺血2 h后,予腹腔内分别注射2、4、6 mg/kg丁苯酞氯化钠注射液,余步骤同IR组。假手术组:栓线插入深度<10 mm,余步骤同IR组。各组于再灌注24 h处死大鼠,分别通过Zealonga法、TTC染色、TUNEL法观察各组大鼠的神经功能缺损评分、脑梗死体积、缺血侧海马CA1区凋亡阳性细胞数,通过免疫组化染色及RT-PCR法观察XIAP和BNIP3阳性细胞数及其mRNA的表达。结果丁苯酞低、中、高剂量亚组神经功能缺损评分均小于IR组(P<0.05),且各亚组之间比较差异有统计学意义(P<0.05)。低剂量亚组脑梗死体积大于中剂量亚组和高剂量亚组(P<0.05)。丁苯酞低、中、高剂量亚组海马CA1区凋亡细胞数逐渐减小,且均小于IR组(P<0.05)。IR组XIAP、BNIP3阳性细胞表达[(22.31±0.94)、(60.13±2.59)个/HP]较假手术组[(3.07±1.43)、(5.78±0.44)个/HP]明显增多(P<0.05);与IR组比较,丁苯酞低、中、高剂量亚组XIAP阳性细胞数[(28.70±1.18)、(32.79±0.88)、(37.01±1.24)个/HP]逐渐增加(P<0.05),BNIP3阳性细胞数[(52.07±1.02)、(40.30±2.00)、(31.04±0.43)个/HP]逐渐减少(P<0.05)。与IR组比较,丁苯酞低、中、高剂量亚组XIAP mRNA逐渐增加、BNIP3 mRNA逐渐减少(P<0.05)。结论丁苯酞氯化钠注射液后处理对大鼠IR具有神经保护作用,其机制与影响XIAP、BNIP3的表达有关,且XIAP、BNIP3阳性表达量与丁苯酞氯化钠注射液存在剂量依赖关系。
Objective The purpose of this study was to observe the effects of dl-3n-butylphthalide（ NBP） sodium chloride injection post-processing on the expressions of X-inhibitor of apoptosis（ XIAP） and Bcl-2/adenovirus E1B19 k Da interacting protein 3（ BNIP3） in the hippocampus CA1 neurons of focal cerebral ischemia reperfusion（ IR） rats,and to investigate the brain-protection mechanisms of NBP. Methods A total of 65 adult male SpragueDawley rats were divided into five groups of equal number,sham operation,IR,and low-,medium- and high-dose NBP,according to the random number table. The IR models were established by modified ligation of the middle cerebral artery. The animals in the NBP groups received intra-abdominal injection of NBP at 2,4,and 6 mg/kg,respectively. All the rats were sacrificed at 24 hours after modeling,neurological scores obtained by Zea Longa,the volume of infarction measured by TTC staining,the number of apoptotic cells counted by TUNEL,and the expressions of XIAP and BNIP3 detected by immunohistochemistry and real-time PCR. Results The neural function defect scores were markedly lower in low-,medium- and high-dose NBP groups than in IR model rats（ P〈0. 05）,with statistically significant differences among the three dose groups（ P〈0. 05）. The volume of infarction was remarkably higher in the low-dose than in the medium- and high-dose NBP groups（ P〈0. 05）. The number of apoptotic cells in the hippocampus CA1 neurons was decreased in the NBP groups as compared with the IR models（ P〈0. 05）. The XIAP- and BNIP3-positive cells were significantly increased in the IR model rats as compared with the sham operation group（ [22. 31 ± 0. 94]and [60. 13 ± 2. 59]/ HP vs [3. 07 ± 1. 43]and [5. 78 ± 0. 44]/ HP,P〈0. 05）. In comparison with the IR models,the NBP-treated rats showed a progressively increased number of XIAP-positive cells in low-,medium-,and high-dose groups（ [28. 70 ± 1. 18],[32. 79 ± 0. 88],and [37. 01 ± 1. 24]/ HP）（ P〈0. 05） but a decreased number of BNIP3-positive cells in the three dose groups（ [52. 07 ± 1. 02],[40. 30 ± 2. 00],and [31. 04 ±0. 43]/ HP）（ P〈0. 05）. Similarly,the expression of XIAP mRNA was up-regulated while that of BNIP3 mRNA down-regulated in the NBP treatment groups as compared with the IR model rats,both in a dose-dependent manner（ P〈0. 05）. Conclusion NBP postprocessing has a neuroprotective effect on IR rats,which is associated with its impact on the expressions of XIAP and BNIP3.
出处
《医学研究生学报》
CAS
北大核心
2015年第12期1257-1261,共5页
Journal of Medical Postgraduates
基金
河北省卫生厅医学科学研究课题计划(20130382)
作者简介
医学硕士研究生
通讯作者：李世英，E-mail：lishiying19701970@163．com
