人参皂苷Rg1、胰岛素样生长因子-1及二者合用抗帕金森病作用的机制研究

发布时间：2018-03-26 16:03

本文选题：人参皂苷Rg1　切入点：胰岛素生长因子-1　出处：《青岛大学》2014年硕士论文

【摘要】：人参皂苷Rgl (ginsenoside Rgl)是人参主要的活性成分,具有神经营养和神经保护作用。胰岛素样生长因子-1(insulin-like growth factor1, IGF-1)是脑内非常重要的生长因子,可以促进神经元的分化和存活。机制研究显示Rgl与IGF-Ⅰ的神经保护作用均与IGF-I受体(IGF-R)信号途径有关,但二者合用是否具有协同作用,尚不清楚。本实验应用6-羟基多巴(6-hydroxydopamine,6-OHDA)损伤多巴胺能神经元细胞系SK-N-SH细胞,建立帕金森病(Parkinson's disease,PD)细胞模型,神经毒素1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, MPTP)制备的PD小鼠模型,应用MTT法、免疫组织化学染色法、高效液相色谱法及免疫印迹法等多种评价方法,探讨人参皂苷Rg1、IGF-1及二者合用对多巴胺能神经元的保护作用及其可能机制。实验结果如下： 1.IGF-1可剂量依赖式对抗6-OHDA对SK-N-SH细胞的损伤(P0.05)。Rg1与IGF-1合用具有协同作用(P0.05)。 2.6-OHDA能够明显降低Bcl-2蛋白表达(P0.05)。Rg1、IGF-1及二者合用均可逆转上述变化,且Rgl与IGF-1合用组作用最明显(P0.05)。 3.6-OHDA可明显升高Bax蛋白表达(P0.05)。Rg1、IGF-1及二者合用均可逆转上述变化(P0.01。 Rg1与IGF-1合用组与单用组相比,无统计学意义(P0.05)。 4. MPTP可明显降低小鼠纹状体DA及其代谢产物DOPAC, HVA的含量,IGF-1(0.1μg/μl,0.5μg/μl,1μg/μl)能够对抗MPTP的毒性作用,其中0.5μg/μl IGF-1神经保护作用最为明显(P0.05)。 5. Rg1、IGF-1及二者合用均可以明显增加DA及其代谢产物DOPAC, HVA含量,与MPTP组相比,差异显著。且Rgl与IGF-1二者合用的效果最为明显(P0.05)。 6. MPTP可明显减少小鼠黑质酪氨酸羟化酶免疫阳性(TH-IR)神经元数目。Rg1、IGF-1及二者合用均可明显增加TH-IR神经元的数目,且以二者合用组的作用最为显著(P0.05)。 7. MPTP可明显减少小鼠黑质抗凋亡蛋白Bcl-2的表达,增加促凋亡蛋白Bax的表达。Rg1、IGF-1及二者合用均可明显逆转上述变化,且以二者合用的效果最为明显(P0.05)。上述结果表明Rg1、IGF-1及二者合用均能够对抗神经毒素对DA神经元的毒性作用,且Rg1与IGF-1合用具有协同作用。本研究为PD的预防与治疗开辟了新的途径,提供了新的实验依据。
[Abstract]:Ginsenoside Rgl (ginsenoside Rgl) is the main active ingredient of ginseng, has neuroprotective effect of insulin-like growth factor -1 (insulin-like growth factor1, IGF-1) is an important growth factor in the brain, can promote neuronal differentiation and survival mechanisms. Studies show neuroprotective effect of Rgl and IGF- I were with the IGF-I receptor (IGF-R) pathway, but the combination of the two will have a synergistic effect, is not clear. The experiment used 6- 6-hydroxydopamine (6-hydroxydopamine, 6-OHDA) SK-N-SH cell injury of dopaminergic neuronal cell lines, the establishment of Parkinson's disease (Parkinson's disease, PD) cell model of neurotoxin 1- methyl -4- phenyl -1,2,3,6- tetrahydropyridine four (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, MPTP) PD mice model preparation, using MTT method, immunohistochemistry, immune method and high performance liquid chromatography A variety of evaluation methods of imprinting, ginsenoside Rg1, IGF-1 and the combination of the two protective effect on dopaminergic neurons and its possible mechanism. The experimental results are as follows:
1.IGF-1 in a dose-dependent manner against 6-OHDA injury in SK-N-SH cells (P0.05) of.Rg1 and IGF-1 has a synergistic effect (P0.05).
2.6-OHDA can decrease the expression of Bcl-2 (P0.05).Rg1, IGF-1 and the combination of the two can reverse these changes, and the Rgl and IGF-1 group had significant effect (P0.05).
3.6-OHDA can significantly increase the expression of Bax (P0.05).Rg1, IGF-1 and the combination of the two can reverse the above changes (P0.01. Rg1 combined with IGF-1 group and single group compared with no statistical significance (P0.05).
4. MPTP can significantly reduce the mouse striatal DA and its metabolites DOPAC, HVA content, IGF-1 (0.1 g/ L, 0.5 g/ L, 1 g/ L) to cytotoxicity against MPTP, of which 0.5 mu g/ Mu L IGF-1 neuroprotective effect was most significant (P0.05).
5. Rg1, IGF-1 and the combination of the two can significantly increase DA and its metabolites DOPAC, HVA content, compared with group MPTP. And the Rgl and IGF-1 combination of the two has the most obvious effect (P0.05).
6. MPTP could significantly reduce the tyrosine hydroxylase positive neurons in mice (TH-IR) the number of.Rg1, IGF-1 and the combination of the two can obviously increase the number of TH-IR neurons, and the combination of the two group is the most significant (P0.05).
7. the expression of MPTP can significantly reduce the anti apoptotic protein Bcl-2 in substantia nigra of mice, increase the expression of.Rg1 apoptosis protein Bax, IGF-1 and the combination of the two can be significantly reversed these changes, and the combined effect of the two most obvious (P0.05).
The results showed that Rg1, IGF-1 and the combination of the two can be toxic effect against the neurotoxin on DA neurons, and Rg1 combined with IGF-1 has a synergistic effect. This study provides a new way for the prevention and treatment of PD, provides a new experimental basis.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.5

【参考文献】