神经调节素-1β对脑出血氧化应激损伤及Nrf2-ARE通路的影响及机制研究

发布时间：2018-06-18 13:14

本文选题：神经调节素-1β + 脑出血　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:本研究通过建立大鼠脑出血模型,探讨神经调节素-1β(neuregulin-1β,NRG-1β)对大鼠脑出血神经功能障碍与脑水肿的疗效,并通过检测内源性抗氧化通路Nrf2(核因子相关因子2)/ARE(抗氧化反应元件)及下游关键分子的表达,探索NRG-1β在脑出血中的抗氧化作用机制。方法:成年健康雄性SD大鼠,随机分为假手术组、模型组、治疗组,每组按造模后1d、3d、5d再分为3个亚组。采用立体定位尾状核注射Ⅳ型胶原酶(0.5U/只)建立大鼠脑出血模型,模型组及治疗组注入Ⅳ型胶原酶,假手术组注入等量的灭菌生理盐水。于术后2小时治疗组大鼠经尾静脉注射重组人NRG-1β(2μg/kg)干预治疗,模型组及假手术组给予等体积的0.1mol/L PBS溶液。分别于各观察时间点对各组大鼠进行神经功能缺损评分、干湿重法检测脑含水量,采用免疫组化法、蛋白免疫印迹法(Western blot)检测血肿周围组织Nrf2、HO-1蛋白表达变化。结果:1、神经功能缺损评分:模型组第1天即出现明显的神经功能缺损,第3天达高峰,神经功能缺损最重,第5天仍有明显的神经功能缺损,但较高峰时减轻,模型组与假手术组比较,各时间点差异有统计学意义(P0.01);治疗组运动、感觉、平衡、反射缺损程度减轻,治疗组与模型组比较,在各观察时间点评分均低于模型组,差异有统计学意义(P0.05);2、脑含水量:模型组大鼠脑含水量呈先升高后下降趋势,在造模第1天即出现脑水肿,在第3天达到高峰,后开始下降,但仍高于假手术组。假手术组和模型组比较,各时间点差异有统计学意义(P0.01);在治疗组中,各时间点脑水肿程度都有所减轻,治疗组与模型组比较,各观察时间点差异具有统计学意义(P0.05)。3、免疫组化检测结果:模型组Nrf2、HO-1于造模后第1天开始升高,在第3天达高峰,而后下降,但表达仍高于假手术组。模型组与假手术组比较,各时间点差异具有统计学意义(P0.05)。治疗组各时间点蛋白表达较模型组升高,治疗组与模型组比较,差异具有统计学差异(P0.05)。4、Western Blot检测结果:模型组在第1天即检测出Nrf2、HO-1蛋白表达升高,第3天达高峰,第5天较峰值有所回落,模型组与假手术组比较,差异有统计学意义(P0.05)。治疗组各时间点Nrf2、HO-1蛋白的表达较模型组均有明显的升高,差异有统计学意义(P0.05)。结论:在大鼠脑出血模型中,Nrf2-ARE通路被激活,NRG-1β的干预可以进一步激活Nrf2-ARE通路,上调下游抗氧化酶表达,发挥抗氧化应激作用。此外,NRG-1β的干预,显著改善脑水肿的程度和神经功能缺损。这些结果表明NRG-1β有效的减轻脑出血继发性损伤。这种神经保护作用可能的机制或许与激活Nrf2-ARE通路相关的抗氧化作用有关。
[Abstract]:Objective: To explore the effect of neuregulin-1 beta (NRG-1 beta) on cerebral hemorrhage and cerebral edema in rats by establishing a rat model of cerebral hemorrhage, and to explore NRG-1 beta by detecting the expression of endogenous antioxidant pathway Nrf2 (nuclear factor related factor 2) /ARE (anti oxygenation element) and the key molecules in the lower reaches of the rat. Methods: antioxidant mechanism in cerebral hemorrhage. Methods: adult healthy male SD rats were randomly divided into sham operation group, model group, and treatment group. Each group was divided into 3 subgroups by 1D, 3D, 5D. The rat model of cerebral hemorrhage was established by injection of type IV collagenase (0.5U/) in the caudate nucleus, and the model group and the treatment group were injected with type IV collagenase, artificial hand. The treatment group was injected with the same amount of sterilized physiological saline. 2 hours after the operation, the rats were injected with recombinant human NRG-1 beta (2 g/kg), the model group and the sham operation group were given equal volume of 0.1mol/L PBS solution. Immunoblotting and protein immunoblotting (Western blot) were used to detect the changes of Nrf2 and HO-1 protein expression around hematoma. Results: 1, nerve function defect score: the model group had obvious nerve function defect in first days, third Tianda peak, the nerve function defect was the heaviest, and the nerve function defect was still obvious in fifth days, but decreased in peak time. Compared with sham operation group, the difference of time points was statistically significant (P0.01). The exercise, sensation, balance, and the degree of reflex defect were reduced in the treatment group. Compared with the model group, the scores of the treatment group and the model group were lower than those in the model group. The difference was statistically significant (P0.05); 2, the brain water content of the model group increased first and then declined. The brain edema appeared at first days and reached the peak in the third day and then began to decline, but it was still higher than the sham operation group. The difference between the sham operation group and the model group was statistically significant (P0.01). In the treatment group, the degree of brain edema at each time point decreased, and the treatment group and the model group were compared with the model group. Study significance (P0.05).3, immunohistochemical detection results: the model group Nrf2, HO-1 began to rise at first days after the model, at the peak of third Tianda, and then decreased, but the expression was still higher than the sham operation group. The difference between the model group and the sham operation group was statistically significant (P0.05). The expression of protein in the treatment group was higher than that in the model group. Compared with the model group, the difference was statistically significant (P0.05).4, Western Blot detection results: the model group detected Nrf2, the expression of HO-1 protein, the peak of Tianda, the peak of third in Tianda, the peak of the peak, and the difference between the model group and the sham operation group was statistically significant (P0.05). The expression of Nrf2 and the expression of HO-1 protein at each time point in the treatment group. Compared with the model group, the difference was statistically significant (P0.05). Conclusion: in the rat model of cerebral hemorrhage, the Nrf2-ARE pathway is activated. The intervention of NRG-1 beta can further activate the Nrf2-ARE pathway, up-regulation the expression of antioxidant enzymes and exert antioxidant stress. In addition, the intervention of NRG-1 beta significantly improves the degree and nerve of brain edema. These results suggest that NRG-1 beta is effective in reducing secondary damage to cerebral hemorrhage. The possible mechanism of this neuroprotective effect may be associated with the antioxidant activity associated with the activation of the Nrf2-ARE pathway.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.34

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