参附注射液对缺氧缺血性脑损伤新生大鼠海马神经元Bax、Bcl-2表达的影响

Effects of Shenfu injection on the expression of Bax and Bcl-2 in the hippocampus neurons of neonatal rat with hypoxic-ischemic brain damage

目的:探讨参附注射液对新生大鼠缺氧缺血性脑损伤(HIBD)后海马神经元Bax、Bcl-2蛋白表达的影响及其治疗作用。方法:建立新生大鼠HIBD模型,把实验分成两部分:①观察缺氧缺血后3、7、14、28天各组的体重变化以及各组28天内生存率;②观察缺氧缺血前2h,缺氧缺血后2、12、24h,3、7、14、28天各时间点新生大鼠海马CA1区神经元Bax、Bcl-2蛋白的表达。每组实验随机分为4组:假手术组(S组)、生理盐水对照组(C组)、参附预处理组(P组)、参附治疗组(SF组)。结果:SF组和P组较C组Bax表达明显下调(P<0.05,P<0.01),而Bcl-2表达显著上调(P<0.05,P<0.01),生长发育和生存情况明显好于C组。结论:参附改善了新生大鼠HIBD后的生长发育,提高了其生存率;可明显下调新生大鼠HIBD后海马神经元Bax蛋白表达及上调Bcl-2蛋白的表达。

Objective： To observe the effect on the expression of Box and Bcl - 2 of neurons in hippocampal CA1 region of neonatal rat with HIBD, Shenfu injection was used to treat HIBD. Methods： The experiment included two parts, one was to observe the effect on the physical development and survival rate, the other was to investigate the expression of Box and Bd -2 of neurons in left hippocampal CA1 region.. Models of postnatal 7 - day Sprague Dawley （SD） rats with HIBD were established. The neonatal rat with H1BD were equally divided into 4 groups： sham operation （S）, control group （C）, Shenfu injection pretreatment （P）, and Shenfu injection treatment （SF）. The in-creasing body weight of rats in every group was observed at 3, 7, 14 and 28 d after HI insult, and the survival rate of rats in every group on 28 d after HI insult. Similarly, four groups were established, however they were further divided into several subgroups again according to the time point 2 h before and 2, 12, 24 h, 3,7, 14 and 28 d after HI insult （ n = 6）. After treatment with Shenfu injection, the expression of Bax and Bcl - 2 in left hippocampal CA1 region at different time was investigated. The expression of Bax and Bcl - 2 was performed by immunohistochemistry. Results： The increase of body weight in group C at 3, 7, 14 and 28 d after HI insult were significantly less than those of group S （P 〈 0. 01 ） and also were group P and SF （P 〈 0. 01, P 〈0. 05）. The increase of body weight in group P at 3, 7, 14 and 28 d after HI insult were significantly higher than those of group C （P 〈 0. 01, P 〈 0. 05）, and the increase of body weight in group SF at 7, 14 and 28 d after HI insult were significantly higher than those of group C （P 〈 0. 05）. The survival rate of group C （60%） was significantly less than that of other groups （P 〈 0. 05）, and there was no obvious difference in survival rates between other groups （ P 〉 0. 05 ）. The increasing expression of Bax and Bcl - 2 in hippocampal CA1 region in group C were observed at 2 h after insult, and at 12 h after insult, compared to group S, the expression of Bax or Bc1-2 in group P was markedly lower or higher than that of group C （P 〈0. 01 ）. The degree of expression were reached the peak at 1 d after insult, and the expression of Bax or Bc1-2 in group P was still markedly lower or higher than that of group C （P 〈0. 01 ）. At 3 d after HI the expression of Bax and Bcl -2 was gradually decreasing, but the expression of Bax or Bcl -2 in group P and SF was markedly lower or higher than that of group C （P 〈 0. 01 ）. At 7 d after HI, the expression of Bax and Bcl -2 was decreased markedly, but the expression of Bax or Bcl- 2 in group C was markedly higher or lower than that of group P and group SF （P 〈 0. 01 ） . Conclusion： Shenfu injection could enhance the physical development and elevate the survival rate in neonatal rats with HI insult. It sigaaiflcantly prevents apoptosis of hippocampus neurons from HI insult by down - regulation of the pro - apoptotic gene Bax, otherwise, and up - regulation of the anti - apoptotic gene Bcl - 2.