| 摘要: | 冠狀動脈疾病是指，因為供給心臟血液和養分的冠狀動脈，產生了狹窄或阻塞，導致心肌損傷所造成的心臟疾病。冠狀動脈疾病是最常見的心臟血
管疾病。Apolipoprotein是脂蛋白分子中的重要組成成分，而Apolipoprotein AI
( Apo-AI)則是高密度脂蛋白(HDL)中的主要結構蛋白。其意義和HDL相同，代表防止血管硬化功能的指標。當濃度愈低意味著清除血管中膽固醇的能力愈差，發生心血管疾病阻塞的風險愈高。本次實驗，以開發組(Discovery set)，利用西方墨點法、奈米級液相層析串聯質譜與生物資訊軟體PEAKS 7分析ApoAI胜肽片段在MDA修飾和轉譯後，在健康人和不同程度冠狀動脈阻塞率的冠狀動脈疾病患者血漿中的表現。再以驗證組(Validation set) 進行: (1)西方墨點法驗證血漿中載脂蛋白AI的表現量。(2)研究血漿中載脂蛋白AI丙二醛修飾胜肽片段中的自體抗體，是否會引發自體免疫反應。並觀察冠狀動脈疾病患者自體抗體是否會因為冠狀動脈阻塞率的嚴重程度而有所影響。(3)檢測血漿中丙二醛加合物(MDA Adduct)，探討冠狀動脈疾病患者冠狀動脈阻塞率的嚴重程度與丙二醛修飾的關係。(4)檢測丙二醛中TBARs的濃度在血漿載脂蛋白AI的丙二醛修飾中冠狀動脈疾病的致病變化。
實驗的結果顯示：(1)健康人與冠狀動脈疾病患者血漿中，ApoAI的表現量沒有顯著的差異(p<0.831)只有CAD+患者血漿中ApoAI的表現量增加0.92倍，ApoAI表現量在CAD-與CAD+的冠狀動脈疾病患者相比較，無顯著差異(p=0.422)但是CAD-與CAD+相比較CAD+的患者增加了0.76倍。在IP-Western實驗後呈現的結果，在Input、Discovery set、Validation set的ApoAI位置，成功被MDA的抗體所辯識。(2)ApoAI的丙二醛修飾胜肽片段在自體抗體(IgG、IgM及IgA)在丙二醛修飾後，CAD-與CAD+的相比，並無顯著的差異。(3)MDA Adduct在冠狀動脈疾病患者的血漿中CAD-與CAD+無顯著的差異，p=0.576。因為本次實驗樣本數有限，期待在未來希望能擴大樣本數並且針對冠狀動脈阻塞率嚴重程度，來探討丙二醛轉譯後修飾載脂蛋白AI與冠狀動脈疾病間的關聯性。
Coronary artery disease refers, because the supply of blood and nutrients coronary heart, resulting in a narrowed or blocked, leading to myocardial damage caused by heart disease. Coronary heart disease is the most common blood
Vascular disease. Apolipoprotein are important components, and Apolipoprotein AI (Apo-AI) is the major structural protein a high density lipoprotein (HDL) . Its meaning and the same HDL, on behalf of indicators to prevent hardening of the arteries function. When the concentration of the lower means the ability to remove cholesterol in the blood vessels worse, the risk of cardiovascular disease, the higher the obstruction. The experiment to develop group (Discovery set), the use of western blot, nanoscale liquid chromatography tandem mass spectrometry and bioinformatics analysis software PEAKS 7 ApoAI peptide fragment after MDA modification and translation in healthy people and different coronary artery disease plasma levels of expression of coronary occlusion rate. And then to verify the group (Validation set): (1) western blot verification plasma apolipoprotein AI performance amount. (2) study of apolipoprotein AI plasma malondialdehyde-modified peptide fragment of autoantibodies, will trigger an autoimmune response. And observe patients with coronary artery disease autoantibodies whether because of the severity of coronary artery occlusion rate be affected. (3) plasma malondialdehyde adducts (MDA Adduct), explore the coronary occlusion rate the severity of coronary artery disease and malondialdehyde-modified relationship. (4) detecting the concentration of MDA in TBARs plasma apolipoprotein AI of malondialdehyde-modified pathogenic changes in coronary artery disease. The experimental results showed that: (1) healthy blood plasma of patients with coronary artery disease, there is no significant difference in the amount of ApoAI of performance (p <0.831) only CAD + plasma of patients show an increase in the amount of 0.92 times ApoAI, ApoAI Performance Measures in CAD- Compared with CAD + patients with coronary artery disease in patients with no significant difference (p = 0.422) but CAD¯ compared with CAD + increased 0.76-fold. Results After IP-Western experiments presented in ApoAI position Input, Discovery set, Validation set and successfully caught antibody of MDA. (2) The difference compared to no significant autoantibodies (IgG, IgM and IgA) after malondialdehyde-modified, CAD- and CAD + of ApoAI of malondialdehyde-modified peptide fragments. (3) MDA Adduct of coronary artery disease in patients with CAD + plasma CAD- no significant difference, p = 0.576. Because the sample size of this experiment is limited, look forward to in the future hopes to expand the number of samples and the severity rate for coronary artery occlusion, to discuss post-translational modification of MDA apolipoprotein AI association between coronary artery disease. |
