清心开窍方对Aβ25-35诱导原代海马神经元损伤IGF-1R、IRS-1及Aβ mRNA表达的影响

Effect of Qingxin Kaiqiao Decoction on Expressions of IGF-1R,IRS-1 and Aβ mRNA in Aβ25-35 Injuried Primary Rat Hippocampus Neurons

目的：研究清心开窍方对Aβ25-35诱导原代海马神经元细胞损伤胰岛素样生长因子1受体（insulin—like growth factor 1 receptor，IGF-1R）、胰岛素受体底物蛋白1（insulin receptor substrate-1，IRS-1）及β-淀粉样蛋白（β—amyloid，Aβ）基因表达的影响，探讨清心开窍方对神经元细胞的保护作用及其机制。方法：制备sD乳鼠原代海马神经元细胞，分为6组：正常对照组、模型组、尼莫地平组（1×107mol.L-1）、清心开窍方低、中、高剂量组（6.25、12.5、25mg/mL）。支持培养至第7天，各治疗组分别给予相应药物预处理24h后，模型组及各治疗组加入老化处理的Aβ25-35。（终浓度为10 μmol.L-1）共同作用24h，建立AD细胞模型。正常对照组给予相等体积的培养液继续孵育24h。采用CCK一8法检测各组的细胞活力，RT—PCR法检测各组IGF-1R、IRS-1和Aβ mRNA的表达水平。结果：模型组与正常对照组比较细胞活性明显降低（P〈0.01），而清心开窍方能有效改善细胞活性；RT—PCR法结果显示模型组IGF-1R mRNA明显降低（P〈0.01），IRS-1和Aβ mRNA表达均明显增加（P〈0.01），而不同浓度的清心开窍方处理后均能提高IGF-1R mRNA的表达（P〈0.01或P〈0.05），减少IRS-1和Aβ mRNA表达（P〈0.01或P〈0.05）。结论：清心开窍方能减轻β一淀粉样蛋白对神经元细胞的毒性作用，提高细胞存活率，保护海马神经元细胞。

Objective： To study the effect of Qingxin Kaiqiao Recipe on the expressions of IGF1R, IRS1 and AI3 mR- NA in Aβ25_35 injuried primary rats hippocampus neurons and explore the protective effect of Qingxin Kaiqiao Recipe on neurons and its mechanism of anti - AD formation. Methods： Get hippocampus neurons of newborn Sprague - Dawley （SD） rats for vitro culture, and they were divided into 6 groups ： normal model, Nimodipine group, Qingxin Kaiqiao De- coction low, middle and high dose groups（6.25 mg.mL-1 , 12.5 mg.mL-1 ,25mg.mL-1）. Different doses of Qingxin Kaiqiao Decoction groups and Nimodipine group were pre - incubated for 24 h. Then all of groups expect normal group were cultured with Aβ25 -35 （final concentration of 10 μmol.L-1） to establish the neuron cell injury, and the nor- mal group was given an equal volume of culture medium and incubated for 24 h. After 24 h, the cell viability was detec- ted by CCK - 8 method, and the expressions of IGF - 1 R, IRS - 1 and Aβ mRNA were detected by RT - PCR. Results ： Compared with the normal control group, the neurons activity was significantly lower in the model group（ P 〈 0. 01 ）, and Qingxin Kaiqiao Decoction can significantly improve it. The results of RT - PCR showed that the expression of IGF - 1R mRNA was significantly decreased and IRS - 1 and Aβ mRNA were increased in model group compared with those of the normal control group（P 〈0. 01 ）, while the different doses of Qingxin Kaiqiao Decoction groups could increase IGF- 1RmRNA, and reduce the expressions of IRS - 1 and Aβ mRNA（P 〈0. 01 or P 〈0. 05）. Conclusion： Qingxin Qingxin De- coction can alleviate the toxicity of β - amyloid on neuronal ceils, increase cell survival rate and protect hippocampal neu- rons.