【摘要】：阿爾茲海默病是一種神經(jīng)系統(tǒng)退行性疾病,其主要的病理特征是腦中老年斑的沉積和神經(jīng)纖維的纏結,但具體的發(fā)病機制至今尚不明確。隨著人口老齡化進程的加快,患有阿爾茲海默病的人數(shù)也在逐年增多,因此急需開發(fā)出一種藥物或治療方法來減少阿爾茲海默病的發(fā)病幾率,延緩阿爾茲海默病的發(fā)生和發(fā)展。目前治療阿爾茲海默病的藥物主要是膽堿酯酶抑制劑,這類藥物只能緩解病人的癥狀,不能達到治療根本的目的,因此科研工作者正致力于開發(fā)能夠治療阿爾茲海默病的新藥物。已有大量的研究表明,具有抗氧化效果的天然產(chǎn)物可能具有治療阿爾茲海默病的作用,天然產(chǎn)物作為源于自然受益自身的藥物,具有良好的應用前景,因此本論文對天然產(chǎn)物進行了抗阿爾茲海默病的藥物篩選。雖然阿爾茲海默病的發(fā)病機制尚不明確,但有幾種假說受到大眾的認可,尤其是Aβ級聯(lián)假說,因此我們所選用的藥物篩選模型為秀麗隱桿線蟲(Caenorhabditis elegans,C.elegans)的Aβ模型CL4176轉(zhuǎn)基因線蟲株,針對Aβ假說篩選抗阿爾茲海默病的藥物。經(jīng)過一系列的篩選我們發(fā)現(xiàn),多種天然產(chǎn)物都具有較好的效果,我們選擇紫娟普洱茶粉作為主要的研究對象。實驗設計了三個給藥濃度,分別為0.1mg/ml、0.2mg/ml和0.4mg/ml,對于CL4176轉(zhuǎn)基因線蟲,三個濃度都具有延緩線蟲癱瘓的作用,并且呈現(xiàn)濃度依賴,因此選擇0.4mg/ml的紫娟普洱茶粉進行機制的研究,對EGCG和原花青素的效果進行驗證發(fā)現(xiàn)0.4mg/ml的紫娟普洱茶粉中發(fā)揮主要作用的不是上述兩種成分。首先對紫娟普洱茶粉發(fā)揮作用的水平進行檢測,發(fā)現(xiàn)0.4mg/ml的紫娟普洱茶粉不影響CL4176線蟲體內(nèi)Aβm RNA的水平,但是能夠抑制Aβ的沉積,說明藥物是在轉(zhuǎn)錄后水平發(fā)揮作用的。其次對CL4176線蟲進行給藥預處理,發(fā)現(xiàn)給藥組仍然具有延緩線蟲癱瘓的作用,因此對線蟲體內(nèi)同抗逆性相關的一些轉(zhuǎn)錄因子進行qPCR檢測,我們發(fā)現(xiàn)sir2.1有明顯的上調(diào)。再次利用TJ356線蟲追蹤了0.4mg/ml的紫娟普洱茶粉對DAF-16入核的影響,結果發(fā)現(xiàn)給藥后能夠促進DAF-16的核轉(zhuǎn)位,利用SOD-3::GFP共表達的線蟲來進一步確認DAF-16的轉(zhuǎn)錄活性對線蟲抵抗氧化應激的必要性,根據(jù)Aβ會引起線蟲體內(nèi)氧化應激增強,及給藥與否CL4176線蟲體內(nèi)ROS的含量及SOD活性的變化,我們推測這可能是給藥以后提高了線蟲抵抗氧化應激的能力。最后對藥物的毒性進行檢測,發(fā)現(xiàn)紫娟普洱茶在安全的劑量下能夠發(fā)揮延緩線蟲癱瘓的作用。綜上所述,本論文通過秀麗隱桿線蟲篩選了抗阿爾茲海默病的藥物,紫娟普洱茶粉能夠抑制Aβ毒性,顯著延緩線蟲的癱瘓,并且這種抑制作用可能是與提高線蟲抵抗Aβ引起的氧化應激的能力和抑制Aβ的沉積有關。
[Abstract]:Alzheimer's disease is a kind of neurodegenerative disease. The main pathological features of Alzheimer's disease are the deposition of senile plaques and the tangles of nerve fibers in the brain. However, the mechanism of Alzheimer's disease is still unclear. With the acceleration of population aging, the number of people suffering from Alzheimer's disease is also increasing year by year. Therefore, it is urgent to develop a drug or treatment to reduce the incidence of Alzheimer's disease and delay the occurrence and development of Alzheimer's disease. The current treatment of Alzheimer's disease is mainly cholinesterase inhibitors, such drugs can only alleviate the symptoms of patients, can not achieve the basic purpose of treatment. So researchers are working on new drugs that can treat Alzheimer's disease. A large number of studies have shown that natural products with antioxidant effects may have the effect of treating Alzheimer's disease, and natural products, as drugs derived from natural benefits themselves, have a good prospect of application. Therefore, natural products were screened for anti-Alzheimer's disease in this paper. Although the pathogenesis of Alzheimer's disease is still unclear, several hypotheses, especially the A 尾 cascade hypothesis, have been accepted by the public. Therefore, the drug screening model we selected is A 尾 CL4176 transgenic nematode strain of Caenorhabditis elegans,C.elegans. Screening of drugs against Alzheimer's disease based on A 尾 hypothesis. After a series of screening, we found that a variety of natural products have a better effect, we choose Zijuan Puer tea powder as the main research object. Three dosages were designed, 0.1 mg / ml 0.2 mg / ml and 0.4 mg / ml respectively. For CL4176 transgenic nematodes, the three concentrations could delay the paralysis of nematodes and present a concentration-dependent manner. Therefore, the mechanism of 0.4mg/ml powder was studied. The effects of EGCG and proanthocyanidins were verified. It was found that the main components of 0.4mg/ml were not the two components mentioned above. At first, the effect level of Puer tea powder from 0.4mg/ml was detected, and it was found that 0.4mg/ml did not affect the level of A 尾 m RNA in CL4176 nematode, but could inhibit the deposition of A 尾, indicating that the drug acted on the posttranscriptional level. Secondly, the pretreatment of CL4176 nematodes showed that the treatment group still had the effect of delaying the paralysis of nematodes. Therefore, some transcription factors related to stress resistance in nematodes were detected by qPCR. We found that sir2.1 was upregulated obviously. TJ356 nematode was used to track the effect of 0.4mg/ml 's Pu'er tea powder on the nucleation of DAF-16. The results showed that the nucleation of DAF-16 was promoted after administration. SOD-3::GFP co-expressed nematodes were used to further confirm the necessity of DAF-16 transcriptional activity for nematode resistance to oxidative stress. According to the effect of A 尾 on the increase of oxidative stress in nematode, the content of ROS and the change of SOD activity in CL4176 nematodes were determined. We speculate that this may increase the ability of nematodes to resist oxidative stress after administration. Finally, the toxicity of Pu'er tea was detected, and it was found that Zijuan Pu'er tea could delay the paralysis of nematodes in a safe dose. To sum up, the drug against Alzheimer's disease was screened by Rhizoma elegans. The tea powder of Zjuanjuan Pu'er can inhibit A 尾 toxicity and delay the paralysis of nematodes. The inhibition may be related to the ability of nematode to resist the oxidative stress induced by A 尾 and to inhibit the deposition of A 尾.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5;R-332

【相似文獻】

相關期刊論文 前10條

1 姚佳紅,李輝;阿爾茲海默病危險因素的病例對照研究[J];中國公共衛(wèi)生;2002年02期

2 劉坤梅,丁宛瓊;早發(fā)家族阿爾茲海默病1個家系報道[J];職業(yè)衛(wèi)生與病傷;2002年04期

3 李電東;第九屆國際阿爾茲海默病會議在美國費城召開[J];中國新藥雜志;2004年08期

4 孫磊;楊瑩;竇彩艷;;阿爾茲海默病治療的研究進展[J];醫(yī)學信息(中旬刊);2010年07期

5 王艷平;翟靜波;朱芳;張雯雯;楊曉娟;曲成毅;;社區(qū)老年人阿爾茲海默病患病及影響因素分析[J];中國公共衛(wèi)生;2011年07期

6 翟月;;線粒體平衡在阿爾茲海默病中的作用[J];中國科技信息;2012年10期

7 張于;程偉;;中西醫(yī)治療阿爾茲海默病的研究現(xiàn)狀[J];中醫(yī)臨床研究;2012年13期

8 鄭亞楠;甘小榮;;我國阿爾茲海默病相關心理學研究綜述[J];中國實用醫(yī)藥;2013年08期

9 謝嵐;艾華;;運動對阿爾茲海默病影響的研究進展[J];中國運動醫(yī)學雜志;2013年09期

10 左萍萍;β淀粉樣肽代謝與阿爾茲海默病[J];標記免疫分析與臨床;2001年01期

相關會議論文 前6條

1 苗雅;何婷;鐘遠;;阿爾茲海默病與磷酸二酯酶4相關性的研究進展(綜述)[A];第三屆江浙滬三地老年醫(yī)學高峰論壇暨2012年浙江省老年醫(yī)學學術年會論文集[C];2012年

2 曼淑梅;陳譽華;;阿爾茲海默病人外周血免疫細胞穿過血腦屏障能力分析[A];中國細胞生物學學會第八屆會員代表大會暨學術大會論文摘要集[C];2003年

3 孫一_，

本文編號：2223021