馬錢苷對MPTP誘導(dǎo)帕金森病小鼠模型的神經(jīng)保護作用
發(fā)布時間：2018-09-07 16:46
【摘要】：帕金森病(Parkinson’s disease,PD)是僅次于阿爾茨海默疾病的第二大神經(jīng)退行性疾病,主要臨床表現(xiàn)為靜止性震顫,行動遲緩為主的運動障礙。目前,對于帕金森病病因的了解并不完全清楚,較多的研究是從環(huán)境、基因和衰老方面對其致病原因進行機制探討,比如炎癥、氧化應(yīng)激,內(nèi)質(zhì)網(wǎng)應(yīng)激,自噬等方面。然而對其治療手段研究的進展較為緩慢,且關(guān)于治療方面的探索力度仍不夠,很多具有潛在作用的單體或單體群有待開發(fā),包括天然產(chǎn)物里面的單體及單體群。馬錢苷(Loganin,Log)是眾多化合物中的一種,為山茱萸(Cornus officinalis sieb.et zuce.)的主要成分之一,已有文獻報道馬錢苷可調(diào)節(jié)大鼠機體免疫反應(yīng);可減輕氧化應(yīng)激損傷;并具有神經(jīng)營養(yǎng)作用,從而提高細(xì)胞的生存能力;改善糖尿病大鼠空間記憶能力。在這里我們選取8周齡左右雄性C57小鼠為實驗動物,采用1-甲基-4-苯基-1,2,3,6四氫吡啶(MPTP)誘導(dǎo)的小鼠帕金森病模型;將給藥組分為馬錢苷預(yù)防給藥組(Log+MPTP)和治療給藥組(MPTP+Log),并設(shè)置預(yù)防對照組(Saline+MPTP)、治療對照組(MPTP+Saline)和空白對照組(Saline)。給藥結(jié)束后用爬桿試驗進行行為學(xué)檢測;用高效液相色譜法(HPLC)檢測小鼠紋狀體內(nèi)多巴胺和3,4-二羥基苯乙酸(DOPAC)的含量變化;同時用western blot、ELISA檢測紋狀體內(nèi)炎癥因子caspase-3、TNF-α,上游調(diào)控因子NFκB,p38、c-Abl,以及與自噬相關(guān)的LC3-II、Drp1表達變化;并結(jié)合免疫熒光染色法檢測膠質(zhì)細(xì)胞和caspase-3、c-Abl、LC3-II腦內(nèi)紋狀體與黑質(zhì)致密部的分布情況;同時采用PC12細(xì)胞建立PD模型后給藥,用吖啶橙染色后觀察酸性自噬泡的水平,以評價馬錢苷對自噬的作用情況。實驗結(jié)果表明,馬錢苷治療給藥組可抑制MPTP誘導(dǎo)的PD小鼠紋狀體內(nèi)多巴胺和TH表達的下降和多巴胺能神經(jīng)元的丟失,且爬桿總運動時間(TLA)下降;但馬錢苷預(yù)防給藥組未見差異。DOPAC的含量在各組間未見差異。經(jīng)過馬錢苷給藥后,小鼠黑質(zhì)內(nèi)小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞的活性較其相應(yīng)對照組有所抑制;caspase-3、TNF-α及其上游調(diào)控因子NFκB,p38和c-Abl的表達較其相應(yīng)對照組均有下降其。提示馬錢苷可能通過抑制炎癥及c-Abl-p38-NFκB信號通路實現(xiàn)保護作用。LC3-II,Drp1表達在馬錢苷給藥后較其對照組也有下降。且細(xì)胞實驗顯示馬錢苷給藥組與其相應(yīng)的對照組相比,細(xì)胞內(nèi)酸性自噬泡水平有所下降,提示馬錢苷也可能對細(xì)胞自噬產(chǎn)生影響。以上研究表明,馬錢苷對MPTP誘導(dǎo)的PD小鼠模型具有神經(jīng)保護的作用,可能是通過抑制膠質(zhì)細(xì)胞的聚集及激活、抑制炎癥因子的表達、并可能通過c-Abl-p38-NFκB通路來抑制細(xì)胞死亡,從而改善行為學(xué)表現(xiàn)。同時馬錢苷也有可能通過影響細(xì)胞自噬來增強細(xì)胞的生存能力。這些結(jié)果為臨床治療帕金森病提供了新的思路。
[Abstract]:Parkinson's disease (Parkinson's disease,PD) is the second most common neurodegenerative disease after Alzheimer's disease. At present, the etiology of Parkinson's disease is not fully understood. Many researches have been done on the pathogenesis of Parkinson's disease from the aspects of environment, gene and aging, such as inflammation, oxidative stress, endoplasmic reticulum stress, autophagy and so on. However, the research on the therapeutic methods is slow, and the research on the treatment is not enough. Many potential monomers or groups of monomers, including those in natural products, need to be developed. Loganin,Log is one of a variety of compounds, which is the (Cornus officinalis sieb.et zuce. of Cornus officinalis. One of the main components has been reported in the literature can regulate the immune response of rats, reduce oxidative stress injury, and neurotrophic effect, thereby improve the viability of cells, improve the ability of spatial memory in diabetic rats. In this paper, we selected male C57 mice about 8 weeks old as experimental animals and used 1-methyl-4-phenyl-1-trihydropyridine 6-tetrahydropyridine (MPTP) to induce the model of Parkinson's disease in mice. The drug groups were divided into two groups: (Log MPTP) in the preventive administration group and (MPTP Log), in the treatment group and (Saline MPTP), in the preventive control group and (Saline). In the blank control group. At the end of administration, the behavioral tests were carried out by climbing pole test, and the contents of dopamine and (DOPAC) in the striatum of mice were detected by HPLC (HPLC), and the changes of dopamine and 3-dihydroxyphenylacetate (DOPAC) in the striatum of mice were determined by HPLC. At the same time, western blot,ELISA was used to detect the expression of inflammatory factor caspase-3,TNF- 偽, upstream regulatory factor NF 魏 B, p38 c-Abland the expression of LC3-II,Drp1 associated with autophagy, and the distribution of glial cells and the substantia nigra compact region in caspase-3,c-Abl,LC3-II brain was detected by immunofluorescence staining. At the same time, PC12 cells were used to establish the PD model and the acridine orange staining was used to observe the level of acidic autophagy to evaluate the effect of marganin on autophagy. The results showed that the treatment group could inhibit the decrease of dopamine and TH expression and the loss of dopaminergic neurons in the striatum of PD mice induced by MPTP, and the decrease of total climbing time (TLA). However, there was no difference in the content of DOPAC between the two groups. The activities of microglia and astrocytes in the substantia nigra of mice were inhibited by the administration of marganin, and the expression of caspase-3 tNF- 偽 and its upstream regulatory factor, NF 魏 B, p38 and c-Abl, decreased compared with those of the corresponding control group. These results suggest that the expression of rhiganoside may be inhibited by inhibiting inflammation and c-Abl-p38-NF 魏 B signaling pathway. The expression of LC3-IIp1 may also decrease compared with that of the control group. Compared with the control group, the intracellular acidic autophagy level in the group treated with marganin decreased, which suggested that it might also have an effect on autophagy. These results suggest that the PD mice model induced by MPTP has neuroprotective effect of marganin, which may inhibit the accumulation and activation of glial cells, inhibit the expression of inflammatory factors, and inhibit cell death through the c-Abl-p38-NF 魏 B pathway. In order to improve behavioral performance. At the same time, it is possible to increase the viability of the cells by affecting autophagy. These results provide new ideas for clinical treatment of Parkinson's disease.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R742.5;R-332
[Abstract]:Parkinson's disease (Parkinson's disease,PD) is the second most common neurodegenerative disease after Alzheimer's disease. At present, the etiology of Parkinson's disease is not fully understood. Many researches have been done on the pathogenesis of Parkinson's disease from the aspects of environment, gene and aging, such as inflammation, oxidative stress, endoplasmic reticulum stress, autophagy and so on. However, the research on the therapeutic methods is slow, and the research on the treatment is not enough. Many potential monomers or groups of monomers, including those in natural products, need to be developed. Loganin,Log is one of a variety of compounds, which is the (Cornus officinalis sieb.et zuce. of Cornus officinalis. One of the main components has been reported in the literature can regulate the immune response of rats, reduce oxidative stress injury, and neurotrophic effect, thereby improve the viability of cells, improve the ability of spatial memory in diabetic rats. In this paper, we selected male C57 mice about 8 weeks old as experimental animals and used 1-methyl-4-phenyl-1-trihydropyridine 6-tetrahydropyridine (MPTP) to induce the model of Parkinson's disease in mice. The drug groups were divided into two groups: (Log MPTP) in the preventive administration group and (MPTP Log), in the treatment group and (Saline MPTP), in the preventive control group and (Saline). In the blank control group. At the end of administration, the behavioral tests were carried out by climbing pole test, and the contents of dopamine and (DOPAC) in the striatum of mice were detected by HPLC (HPLC), and the changes of dopamine and 3-dihydroxyphenylacetate (DOPAC) in the striatum of mice were determined by HPLC. At the same time, western blot,ELISA was used to detect the expression of inflammatory factor caspase-3,TNF- 偽, upstream regulatory factor NF 魏 B, p38 c-Abland the expression of LC3-II,Drp1 associated with autophagy, and the distribution of glial cells and the substantia nigra compact region in caspase-3,c-Abl,LC3-II brain was detected by immunofluorescence staining. At the same time, PC12 cells were used to establish the PD model and the acridine orange staining was used to observe the level of acidic autophagy to evaluate the effect of marganin on autophagy. The results showed that the treatment group could inhibit the decrease of dopamine and TH expression and the loss of dopaminergic neurons in the striatum of PD mice induced by MPTP, and the decrease of total climbing time (TLA). However, there was no difference in the content of DOPAC between the two groups. The activities of microglia and astrocytes in the substantia nigra of mice were inhibited by the administration of marganin, and the expression of caspase-3 tNF- 偽 and its upstream regulatory factor, NF 魏 B, p38 and c-Abl, decreased compared with those of the corresponding control group. These results suggest that the expression of rhiganoside may be inhibited by inhibiting inflammation and c-Abl-p38-NF 魏 B signaling pathway. The expression of LC3-IIp1 may also decrease compared with that of the control group. Compared with the control group, the intracellular acidic autophagy level in the group treated with marganin decreased, which suggested that it might also have an effect on autophagy. These results suggest that the PD mice model induced by MPTP has neuroprotective effect of marganin, which may inhibit the accumulation and activation of glial cells, inhibit the expression of inflammatory factors, and inhibit cell death through the c-Abl-p38-NF 魏 B pathway. In order to improve behavioral performance. At the same time, it is possible to increase the viability of the cells by affecting autophagy. These results provide new ideas for clinical treatment of Parkinson's disease.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R742.5;R-332
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