清脑益元汤对大鼠脑缺血损伤VEGF、PDGF表达的影响

发布时间：2018-06-18 15:00

本文选题：清脑益元汤 + 脑缺血损伤　；参考：《广西中医药大学》2016年硕士论文

【摘要】：目的:观察清脑益元汤对大鼠脑缺血损伤缺血侧皮层及海马区VEGF、PDGF表达的影响,探讨清脑益元汤对大鼠脑缺血损伤神经元保护的作用机制。方法:将成年健康雄性SD大鼠192只,按随机数字表分为两大组,每组96只,继续随机分为4个亚组:空白组(A组6只)、假手术组(B组30只)、模型对照组(C组30只)、清脑益元组(D组30只)。假手术组、模型对照组及清脑益元组均按缺血损伤后1d、3d、7d、14d、28d五个处死时间点分为5个亚组(n=6),并分别作如下处理:空白正常饲养,灌胃等体积蒸馏水;假手术组单纯手术而未插线动物,术后正常饲养,灌胃等体积蒸馏水;模型对照组灌胃等体积蒸馏水7d后,麻醉,行MACO术,术后制成大鼠急性脑缺血损伤大鼠模型,待大鼠麻醉清醒后,继续予生理盐水灌胃至各处死时间点。清脑益元组灌胃等体积清脑益元7d后操作同模型对照组,待大鼠麻醉清醒后,继续予中药灌胃至大鼠各处死时间点。参考Longa的5分制法对大鼠进行神经功能缺损评分,并按各时间点进行标本采集。取材后,一组应用实时定量逆转录聚合酶链反应(Real Time PCR)检测大鼠缺血侧皮层及海马区VEGF的表达,另一组应用蛋白质印迹法(Western Blot)检测大鼠缺血侧皮层及海马区PDGF的表达。结果:1.RT-PCR检测结果:与假手术组比较,模型组脑组织VEGF的mRNA表达在各时间点均有所升高,但统计学比较无显著差异。与模型组比较,清脑益元组VEGF mRNA表达在各时间点均显著增高。清脑益元组早期(第1天)可见VEGF表达增高,在早期(第3天)就明显升高,并逐渐升高第14天达到高峰;在模型对照组VEGF表达成缓慢增长的趋势,差异有统计学意义(P0.05)。2.蛋白质印迹法结果:与模型组比较,清脑益元组PDGF的表达在各时间点均显著增高。清脑益元组早期(第1天)可见PDGF的明显增高,在早期(第3天)呈稍放缓但仍很明显的增高趋势,并逐渐升高第14天达到高峰;在模型组PDGF表达成缓慢增长的趋势,差异有统计学意义(P0.05)。结论:清脑益元汤对大鼠脑缺血损伤神经元的保护作用可能与VEGF、PDGF表达的增加有关。
[Abstract]:Objective: to observe the effect of Qingnao Yiyuan decoction (QNYD) on the expression of VEGF PDGF in ischemic cortex and hippocampus of rats with cerebral ischemia injury, and to explore the protective mechanism of Qingnao Yiyuan decoction (QIYD) on neurons injured by cerebral ischemia in rats. Methods: 192 adult male SD rats were randomly divided into two groups, 96 rats in each group. They were randomly divided into four subgroups: group A (n = 6), group B (n = 30), group C (n = 30) and group D (n = 30). Sham-operation group, model control group and Qingnao Yiyuan group were divided into 5 subgroups according to five death time points: normal blank feeding and gastric distilled water. In the sham operation group, the rats were fed normally and fed with distilled water of the same volume after operation, and the rats in the model control group were anesthetized and treated with MACO for 7 days after the rats were fed with distilled water of the same volume. The model of acute cerebral ischemia injury in rats was made after operation. After the rats were anesthetized and awake, the rats were gavaged with normal saline until all the time points were put to death. The rats in the Qingnao Yiyuan group were treated with the same model control group after 7 days of oral administration of the same volume of Qingnao Yiyuan. After the rats were anesthetized and awake, the rats were continued to be gavaged with traditional Chinese medicine to the death time points of the rats. According to the 5-point method of Longa, the neurological impairment score of rats was evaluated, and the specimens were collected according to the time points. The expression of VEGF in ischemic cortex and hippocampus was detected by real time PCR and Western blot was used to detect the expression of PDGF in ischemic cortex and hippocampus of rats. Results 1. The results of RT-PCR: compared with sham operation group, the expression of VEGF mRNA in brain tissue of model group was increased at all time points, but there was no significant difference between the two groups. Compared with model group, VEGF mRNA expression in Qingnao Yiyuan group was significantly increased at all time points. In Qingnao Yiyuan group, the expression of VEGF increased at the early stage (on the first day), increased obviously at the early stage (on the third day), and gradually increased to the peak on the 14th day, while in the model control group, the expression of VEGF increased slowly, and the difference was statistically significant (P 0.05). Results of Western blotting: compared with model group, PDGF expression in Qingnao Yiyuan group was significantly higher than that in model group. In Qingnao Yiyuan group, PDGF increased significantly at the early stage (day 1), but decreased slightly at the early stage (day 3), and reached its peak on the 14th day, and the expression of PDGF increased slowly in the model group. The difference was statistically significant (P 0.05). Conclusion: the protective effect of Qingnao Yiyuan decoction on neurons injured by cerebral ischemia may be related to the increase of VEGF PDGF expression.
【学位授予单位】：广西中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.7

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