β-amyloid誘發SK-N-MC細胞的絲裂原活化蛋白酶磷酸化並用包覆薑黃素之心磷脂微脂粒負載麥胚凝集素抑制阿茲海默症模型的β-amyloid沉積

Phosphorylation of mitogen-activated protein kinases in Sk-N-MC cells and curcumin encapsulated into cardiolipin liposomes grafted with wheat germ agglutinin for inhibition of β-amyloid deposit in Alzheimer’s disease models

林哲宇(Zhe-Yu Lin)

指導教授 ： 郭勇志

國立中正大學/工學院/化學工程學系/碩士(2015年)

摘要

本研究將cardiolipin (CL)組成的微脂粒(liposomes)包覆薑黃素(curcumin, CRM)和神經成長因子(nerve growth factor, NGF)做為藥物載體，以麥胚凝集素(wheat germ agglutinin,WGA)接枝於載體表面，並探討CL對蛋白質磷酸化的影響。實驗中利用β-Amyloid (Aβ)誘發人類神經胚胎腫瘤細胞株SK-N-MC cells的蛋白質磷酸化，建立一個體外模型來模擬阿茲海默症(Alzheimer’s disease, AD)蛋白質磷酸化的情形。實驗結果顯示Aβ誘發SK-N-MC細胞的p38和JNK蛋白磷酸化(p-p38: 0.8; p-JNK:0.9)明顯高於控制組(p-p38: 0.48; p-JNK: 0.38 )，對於治療AD體外模型的研究結果顯示包覆薑黃素的微脂粒能有效抑制p38、JNK和S202蛋白磷酸化，減緩細胞凋亡的現象。含有CL的微脂粒包覆神經成長因子能有效的提高p-ERK5的表現量(Aβ: 0.98; Aβ+NGF-CL/LIP: 1.56)，達到能使細胞存活的效果。p-38和p-JNK的免疫螢光染色指出用10 μM的Aβ處置SK-N-MC細胞24 h後，有達到細胞凋亡的效果。
動物實驗中發現，直接將Aβ注入在Wistar大鼠的海馬迴CA1區域，經過2個星期後能確實看到Aβ沉積以及斑塊的形成，實驗結果顯示，經由WGA-CL微脂粒的治療，能減少Aβ斑塊的面積和脂質過氧化的表現量(control: 0.374 nmol/mg; AD: 2.5 nmol/mg; AD+WGA-CRM-CL/LIP: 0.85 nmol/mg)，以及保護尼氏體(Nissl body)不被降解掉，故可應用於未來阿茲海默症的治療。

關鍵字

阿茲海默症 ； β型類澱粉蛋白 ； MAPK路徑 ； 磷酸化 ； 薑黃素 ； 神經成長因子

並列摘要

Curcumin and nerve growth factor (NGF) encapsulated into cardiolipin (CL) liposomes affeced phosphorylation of protein in this reserch. We used β-Amyloid (Aβ) to induce phophorylation of protein in SK-N-MC cells for simulating Alzheimer’s disease (AD). Our results suggest that Aβ induced level of phosphorylation is more than control groups and curcumin encapsulated into cardiolipin liposomes inhibit significantly phosphorylation of p38, JNK and S202. NGF encapsulated into cardiolipin liposomes can increase p-ERK5 expression significantly. Immunocytochemistry of p-p38 and p-JNK indicates apoptosis for treating SK-N-MC cells with 10 μM Aβ for 24 h. Futhermore, in an AD rat model by the hippocampus injection of Aβ, WGA-CL liposomes significantly decrease Aβ plaques area and level of lipid peroxidation. WGA-CRM-CL liposomes also can protect Nissl bodies against Aβ-induced degeneration. According to our results, we conclude that WGA-CL liposomes can be a useful delivery system for AD treatment in preclinical trials.

並列關鍵字

curcumin ； Alzheihmer’s disease ； β-amyloid ； MAPK pathway ； phosphorylation ； NGF

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