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目的:观察清热化瘀Ⅱ号方对脑缺血再灌注损伤大鼠TLR4、TRIF mRNA表达的影响。方法:将成年SD大鼠随机分为两大组,每组再分为:空白对照组(n=6)、假手术组(n=30)、模型组(n=30)、清热化瘀Ⅱ号方组(n=30)。采用线栓法阻断大脑中动脉制备I/R模型,除空白对照组外,每组按照缺血2h后再灌注3、6、12、24、48h 5个时间点分为5个亚组。取材后将一组脑组织行常规HE染色并在光学显微镜下观察其病理形态学改变,另一大组取材后采用实时荧光定量PCR法观察各个时间点TLR4、TRIF mRNA的表达变化。结果:1HE染色结果:光镜下观察清热化瘀Ⅱ号方组与模型组的脑组织病理形态学比较,在各个时间点神经元受损明显减轻。2荧光定量PCR反应结果:TLR4、TRIF mRNA在空白对照组、假手术组均有少量表达,模型组缺血各时间点其表达含量较其他组均明显增多(P<0.05);模型组大鼠随再灌注时间的延长TLR4、TRIF mRNA的表达含量逐渐升高,脑缺血再灌注24h后其mRNA表达达高峰,48h后开始下降;清热化瘀Ⅱ号方组较模型组各时间点其表达含量显著降低,差异均有统计学意义(P<0.05)。结论:清热化瘀Ⅱ号方可通过降低TLR4、TRIF mRNA的表达,以减少神经元的凋亡、炎性反应及脑组织受损程度进而改善脑内炎症环境,促进脑缺血再灌注损伤后的神经功能恢复。
Objective: To observe the effect of Qingre Huayu Recipe (Ⅱ) on TLR4 and TRIF mRNA expression in rats with cerebral ischemia-reperfusion injury. Methods: Adult SD rats were randomly divided into two groups, each group was divided into blank control group (n = 6), sham operation group (n = 30), model group (n = 30), Qingre Huayu Ⅱ Square group (n = 30). The occlusion of middle cerebral artery by thread occlusion was used to prepare the I / R model. Except for the blank control group, each group was divided into 5 subgroups according to the time points of 2h, 2h, 3h, 12h, 24h and 48h after reperfusion. A group of brain tissues were harvested routinely and stained with HE for histopathological examination under light microscope. The other group was observed by real-time fluorescence quantitative PCR to observe the expression of TLR4 and TRIF mRNA at different time points. Results: The results of 1HE staining: Compared with the model group, the histopathology of Qingre Huayu Ⅱ group and the model group were observed under the light microscope. The neuron damage was obviously alleviated at each time point. 2 Fluorescence quantitative PCR results: TLR4, TRIF mRNA in the blank control group and sham operation group were a small amount of expression, the expression of the model group at each time point were significantly increased compared with other groups (P <0.05); model group rats The expression of TRIF mRNA increased gradually with the reperfusion time prolonged, and the mRNA expression of TRIF reached the peak at 24 hours after cerebral ischemia-reperfusion, and then decreased at 48 hours. The expression of TRIF mRNA in Qingre Huayu Ⅱ group was higher than that of the model group at each time point Content was significantly lower, the difference was statistically significant (P <0.05). Conclusion: Qingre Huayu Decoction can improve the inflammatory environment in the brain and promote the cerebral ischemia-reperfusion injury by decreasing the expression of TLR4 and TRIF mRNA to reduce the neuronal apoptosis, inflammatory reaction and brain tissue damage Nerve function recovery.