丁苯酞氯化钠注射液后处理对局灶性脑缺血再灌注大鼠海马CA1区凋亡相关因子表达的影响

发布时间：2018-10-19 16:00

【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(P0.05),且各亚组之间比较差异有统计学意义(P0.05)。低剂量亚组脑梗死体积大于中剂量亚组和高剂量亚组(P0.05)。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(P0.05)。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(P0.05);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(P0.05),BNIP3阳性细胞数[(52.07±1.02)、(40.30±2.00)、(31.04±0.43)个/HP]逐渐减少(P0.05)。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(P0.05)。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。
[Abstract]:Objective to investigate the effect of butyphthalide sodium chloride injection on X-linked inhibitor of apoptosis (X-inhibitor of apoptosis,XIAP) and Bcl-2/ adenovirus E1B19k Da interacting protein 3 (Bcl-2/adenovirus E1B19k Da interacting protein _ 3 BNIP3). The purpose of this study was to observe the effect of butyphthalide sodium chloride injection on the expression of XIAP,BNIP3 in the ischemic hippocampus of rats with focal cerebral ischemia-reperfusion injury (ischemia reperfusion,IR) and to explore the brain protective mechanism of butyphthalide sodium chloride injection. Methods Sixty-five clean grade male SD rats were randomly divided into three groups: sham operation group (n = 13), butyphthalide group (n = 39), low dose group, middle dose group and high dose subgroup. 13 rats in each group. IR group: IR model of middle cerebral artery in rats was made by modified thread embolization (ischemia for 2 h, reperfusion for 24 h). The rats in the low, middle and high dose subgroups were given intraperitoneal injection of butyphthalide sodium chloride for 2 mg/kg after ischemia for 2 h. The remaining steps were the same as those in the IR group. Sham operation group: the insertion depth of the thread was 10 mm, and the step was the same as that in the IR group. The rats in each group were killed at 24 h after reperfusion. The neurological deficit score, cerebral infarct volume and the number of apoptotic positive cells in CA1 region of hippocampus of ischemic side were observed by Zealonga, TTC staining and TUNEL method. The number of XIAP and BNIP3 positive cells and the expression of mRNA were observed by immunohistochemical staining and RT-PCR method. Results the neurological deficit scores in the middle and high dose subgroups were lower than those in the IR group (P0.05), and the differences among the subgroups were statistically significant (P0.05). The volume of cerebral infarction in low dose subgroup was larger than that in middle dose subgroup and high dose subgroup (P0.05). The number of apoptotic cells in the hippocampal CA1 region of the high dose subgroup was lower than that in the IR group (P 0.05 卤0.94), (60.13 卤2.59 / HP), and the number of apoptotic cells in the high dose subgroup was significantly higher than that in the sham operation group [(3.07 卤1.43), (5.78 卤0.44) / HP] (P0.05), and was significantly lower than that in the IR group (P 0.05), while the expression of XIAP,BNIP3 positive cells in the IR group (P 0.05 卤0.94), (60.13 卤2.59) was significantly higher than that in the sham operation group (P 0.05). In the high dose subgroup, the number of XIAP positive cells [(28.70 卤1.18), (32.79 卤0.88), (37.01 卤1.24) / HP] increased gradually (P0.05), and the number of BNIP3 positive cells [(52.07 卤1.02), (40.30 卤2.00), (31.04 卤0.43) / HP] decreased gradually (P0.05). Compared with IR group, butyphthalide was lower, XIAP mRNA increased gradually and BNIP3 mRNA decreased gradually in middle and high dose subgroups (P0.05). Conclusion Butylphthalide sodium chloride injection has neuroprotective effect on IR in rats, and its mechanism is related to the influence of XIAP,BNIP3 expression, and the positive expression of XIAP,BNIP3 is in a dose-dependent manner with butyphthalide sodium chloride injection.
【作者单位】：华北理工大学附属医院神经内一科;华北理工大学药理学教研室;
【基金】：河北省卫生厅医学科学研究课题计划(20130382)
【分类号】：R965

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