本文選題：杜鵑素 + 小膠質細胞　； 參考：《吉林大學》2017年碩士論文

【摘要】：近年來,大量研究表明神經炎癥在神經退行性疾病的發(fā)生發(fā)展過程中起到重要作用。對死后PD病人剖檢及6-羥多巴胺(6-OHDA)和MPTP誘導的PD動物模型中都發(fā)現(xiàn)中腦黑質區(qū)存在小膠質細胞異常活化的現(xiàn)象。小膠質細胞是神經炎癥反應的主要效應細胞,其數(shù)量僅占膠質細胞總數(shù)的20%[1]。神經炎癥過程中小膠質細胞被過度活化后,能產生一些細胞毒性因子(IL-1β、IL-6、TNF-α、NO和PGE2等),這些細胞毒性因子可以導致周圍的神經元發(fā)生損傷,損傷神經元釋放的損傷相關模式分子可以進一步活化小膠質細胞,從而產生更多細胞毒性因子進一步損傷周圍的神經元[2]。目前已經有研究表明,抗炎藥物能減少PD的患病率。因此,抑制小膠質細胞活化可能對PD起到治療或緩解作用。杜鵑素(Farrerol)是提取自滿山紅或者其他杜鵑屬植物的一種黃酮類化合物,在體內起到抗炎、抗氧化和免疫抑制等多方面的功能[3]。有研究結果顯示[4,5],杜鵑素在疾病中的很多積極作用都與其參與調節(jié)NF-κB通路的激活相關;另外,杜鵑素可通過抑制促炎蛋白酶類(iNOS和COX-2)及其產物NO和PGE2來達到減輕炎癥的作用,所以,我們推測在PD中杜鵑素可能通過抑制神經炎癥反應從而對PD起到治療和緩解作用。因此,本實驗通過建立LPS誘導的體外神經炎癥反應細胞模型和體內PD動物模型,研究了杜鵑素對神經炎癥介導的PD模型的影響及其作用機制。通過對小膠質細胞系BV-2細胞的研究發(fā)現(xiàn),杜鵑素對LPS誘導的炎癥模型中促炎細胞因子(IL-6、IL-1β和TNF-α)及促炎蛋白酶類(iNOS和COX-2)表達具有顯著的抑制作用,且呈劑量依賴性。同時,杜鵑素也可以顯著抑制LPS誘導的BV-2細胞中NO和PGE2產量的增加。另外,通過Western blot方法檢測杜鵑素對LPS誘導的MAPKs、NF-κB和Akt信號通路關鍵節(jié)點磷酸化水平的影響,結果發(fā)現(xiàn)杜鵑素可以顯著抑制LPS誘導的NF-κB p65和Akt的磷酸化水平,但對MAPKs通路中ERK1/2、p38和JNK1/2的磷酸化水平沒有影響。以上結果表明,杜鵑素可以顯著抑制LPS在BV-2細胞中誘導的炎癥反應,其機制可能是通過抑制NF-κB p65和Akt的磷酸化。為了進一步確定杜鵑素的抗神經炎癥功能對PD的影響,我們通過在中腦黑質區(qū)注射LPS建立PD動物模型,并腹腔注射杜鵑素研究其對PD的治療作用。結果顯示,杜鵑素可以顯著抑制阿撲嗎啡誘導的PD動物模型的旋轉行為;機制研究發(fā)現(xiàn),杜鵑素顯著抑制PD動物模型中中腦黑質區(qū)小膠質細胞的激活以及多巴胺能神經元的減少。以上研究結果表明,杜鵑素可以通過抑制NF-κB-p65和Akt的磷酸化改善LPS在小膠質細胞中誘導的炎癥反應;此外,杜鵑素在PD動物模型中通過抑制小膠質細胞介導的神經炎癥反應過程中對多巴胺能神經元起到保護作用。因此,杜鵑素有望成為PD治療的有效藥物。
[Abstract]:In recent years, a large number of studies have shown that neuroinflammation plays an important role in the occurrence and development of neurodegenerative diseases. Abnormal activation of microglia in substantia nigra was found in postmortem PD patients, 6-OHDA (6-OHDA) and MPTP induced PD animal models. Microglial cells are the main effector cells of neuroinflammatory reaction, which only account for 20% of the total number of glial cells [1]. In the process of neuroinflammation, microglia are over-activated and produce some cytotoxic factors (IL-1 尾, IL-6, TNF- 偽, no, PGE2, etc.). These cytokines can cause damage to the surrounding neurons. Injury-related model molecules released by injured neurons can further activate microglia and produce more cytotoxic factors to further damage the surrounding neurons [2]. Studies have shown that anti-inflammatory drugs can reduce the prevalence of PD. Therefore, inhibition of microglial activation may play a therapeutic or remission role in PD. Farrerol is a flavonoid extracted from Rhododendron or other Rhododendron plants. Farrerol has many functions of anti-inflammatory, anti-oxidation and immunosuppressive in vivo [3]. Some studies have shown that Rhododendron is involved in regulating the activation of NF- 魏 B pathway in many of its positive roles in the disease. In addition, rhododendronin can attenuate inflammation by inhibiting proinflammatory proteases (iNOS and COX-2) and their products, no and PGE2. Therefore, we speculate that rhododendron may play a therapeutic and remission role in PD by inhibiting neuroinflammatory response. Therefore, the effects of rhododendron on neuroinflammatory PD model induced by LPS in vitro and PD animal model in vivo were studied. Through the study of microglial cell line BV-2, it was found that rhododendron inhibited the expression of pro-inflammatory cytokines (IL-6, IL-1 尾 and TNF- 偽) and pro-inflammatory proteases (iNOS and COX-2) in a dose-dependent manner. At the same time, rhododendron could significantly inhibit the increase of no and PGE2 production in BV-2 cells induced by LPS. In addition, the effects of rhododendron on LPS-induced phosphorylation of NF- 魏 B and Akt key nodes in MAPK- 魏 B and Akt signaling pathway were detected by Western blot. The results showed that Rhododendron could significantly inhibit the phosphorylation of LPS-induced NF- 魏 B p65 and Akt. But there was no effect on the phosphorylation levels of ERK1 / 2 p38 and JNK1 / 2 in MAPKs pathway. These results suggest that Rhododendron can significantly inhibit the inflammatory response induced by LPS in BV-2 cells by inhibiting the phosphorylation of NF- 魏 B p65 and Akt. In order to further determine the effect of rhododendron on PD, we established PD animal model by injecting LPS into substantia nigra of mesencephalon, and intraperitoneally injected rhododendron to study the therapeutic effect of Rhododendron on PD. The results showed that rhododendron could significantly inhibit the rotational behavior of PD model induced by apomorphine. Rhododendron significantly inhibited the activation of microglia and the decrease of dopaminergic neurons in substantia nigra in PD animal model. These results suggest that Rhododendron can improve the LPS-induced inflammatory response in microglia by inhibiting the phosphorylation of NF- 魏 B-p65 and Akt. Rhododendron protects dopaminergic neurons by inhibiting neuroinflammatory responses mediated by microglia in PD animal models. Therefore, rhododendron is expected to be an effective drug in the treatment of PD.
【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

【參考文獻】

相關博士學位論文 前2條

1 李建寬;杜鵑素對氧化應激誘導血管內皮細胞損傷的保護作用及分子機制研究[D];山西醫(yī)科大學;2014年

2 慈鑫鑫;杜鵑素對卵蛋白和內毒素誘導氣道炎癥的作用及機制研究[D];吉林大學;2012年

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