破血化瘀汤治疗急性脑内血肿大鼠疗效及其对VEGF、GAP-43影响的研究

Effect on VEGF and GAP-43 of Poxue Huayu prescription on acute intracerebral hematoma in rats

目的:探讨破血化瘀汤对急性脑内血肿大鼠的治疗效果及其对VEGF、GAP-43表达的影响。方法:对急性脑内血肿大鼠进行以1.25%、2.5%、5%的破血化瘀汤剂灌胃给药,并通过酶联免疫法、免疫组化和RT-PCR检测VEGF分泌和表达的情况。按5、10、15d再分为三个亚组,每组20只。制备急性脑内血肿动物模型。免疫组化检测各实验组大鼠血肿区皮层GAP-43。结果:2.5%含药血清与2.5%空白对照组相比能显著提高血管内皮生长因子VEGF的含量(P<0.05),而其余浓度的含药血清对VEGF的作用不是十分显著。图像分析结果显示,与同血清含量的空白对照相比,2.5%含药血清组的VEGF灰度值最小(P<0.05),另外两种药物浓度对VEGF的含量与合成均无显著影响。RT-PCR的检测结果显示,2.5%含药血清与2.5%的空白对照组相比,血管内皮生长因子VEGF含量、及其受体的转录水平(P<0.05)二者都显著提高;而较高浓度5%含药血清与5%的空白对照组相比,5%含药血清对VEGF本身的转录有影响;而对VEGF及其受体的表达均无显著影响是较低浓度1.25%含药血清。造模后的第5天,各破血化瘀GAP-43均有表达,随时间增加表达也随之增加,在第10天后,开始有不同程度下降。与空白组相比,破血化瘀汤组10、15d的GAP-43阳性细胞数显著增加,有统计学意义(P<0.05)。结论:破血化瘀剂可上调VEGF途径,说明了药物促血管新生以及利于大脑神经元神经GAP-43再生。

Objective： To explore the therapeutic effect of Poxue Huayu prescription on acute intracerebral hematoma in rats and its effect on VEGF and GAP-43 expression. Methods： Using gastrointestinal administration, 1.25%, 2.5% and 5% of the Poxue Huayu prescription to treat acute cerebral hematoma in rats, and by enzyme-linked immunoassay, immunohistochemistry and RT-P-CR detection of hematoma vascular endothelial growth factor （vascular endot helial growth factor VEGF） and secretion and expression.Divided into 3 subgroups according to 5 days, 10 days and 15 days, 20 in each group.Animal models of acute intracerebral hematoma were prepared.Imm-un-ohistochemical detection of the experimental group rats hematoma cort-ex growth associated protein 43（43）Growth associated protein, GAP-43. Results：2.5% drug-containing serum significantly increased vascular endothelial growth factor VEGF （ P 〈0.05） compared with 2.5% blank control group, while other concentrations of drug-containing serum were not very significant for VEGF. The results of image analysis showed that compared with the serum control group, the VEGF gray value of the 2.5% drug-containing serum group was the smallest （ P 〈0.05）, and the other two drug concentrations had no significant effect on the VEGF content and synthesis. The results of RT-PCR showed that 2.5% of drug-containing serum compared with 2.5% of the blank control group, the vascular endothelial growth factor VEGF content and its receptor transcription level （ P 〈0.05） was significantly improved; Compared with 5% of the blank control group, 5% drug-containing serum had an effect on the transcription of VEGF itself, but had no significant effect on the expression of VEGF and its receptor. The lower concentration was 1.25%. Drug serum. On the 5th day after model establishment, GAP-43 was expressed in all blood stasis and phlegm, and the expression increased with time. After the 10th day, it began to decrease to varying degrees. Compared with the blank group, the number of GAP-43 positive cells in the blood-breaking Huayu decoction group in creased significantly at 10 and 15 days, and the difference was statistically significant （ P 〈0.05）.Conclusion： By upregulation of VEGF pathway, the anticoagulant can promote angiogenesis and promote the regeneration of GAP-43 neurons in brain.