臺灣博碩士論文加值系統

薑黃素 (CM) 是一種天然親油性的多酚物質，具有許多藥理活性，如抗氧化、抗發炎、抗菌、抗病毒、抗風濕、抗癌和保護神經等功能。除了許多益處外，薑黃素的缺點為水溶性低，就連薑黃素衍生物也是。然而，近期有許多研究是利用生物聚合物包覆疏水性物質以改善其生體可用率。因此，本研究目的為評估以玉米醇溶蛋白奈米粒子作為薑黃素及其衍生物口服載體的可行性，並探討其包括CM-1、CM-2、CM-A、CM-F和tetrahydrocurcumin 對ABTS 和DPPH 之自由基清除活性及α-葡萄糖苷酶和黃嘌呤氧化酶之抑制活性。傅立葉紅外線光譜儀亦用於玉米醇溶蛋白-薑黃素奈米粒子的鑑定和特性研究。結果顯示，衍生物中除了CM-1 外，都具備與維生素E 為CM-F > CM > CM-A > CM-1，且都較標準品槲皮素之抑制效果好。此外，只有CM-F 具有良好的黃嘌呤氧化酶抑制活性（IC50 = 2.05 ± 0.15 μM），甚至比異嘌呤醇（IC50 = 12.04 ± 1.54 μM）具有更佳的抑制作用。奈米粒子之FTIR 圖譜觀察到玉米醇溶蛋白amide I 和amide II 及amide III的波峰位置出現或有些微轉移；且化合物本身的幾個主要波峰也發現有存在或轉移的現象出現；證實玉米醇溶蛋白與化合物透過疏水作用區產生部分氫鍵鍵結。製備出無添加界面活性劑的奈米粒子是圓球狀，小的（平均粒徑 ≈ 125 - 150 nm），並具有狹窄的粒徑分布結果（多分散性指數 < 0.4）；此外，透過示差掃描量熱吸法測定，包覆的化合物為非結晶態。綜合上述，已確認玉米醇溶蛋白可當薑黃素及其衍生物的載體。特別是，玉米醇溶蛋白與CM-F 結合之奈米顆粒是用於治療痛風和糖尿病最具潛力的抑制劑。

Curcumin (CM) is a natural lipophilic polyphenol with a variety of pharmacological properties, that provide antioxidizing, antiinflammatory, antimicrobial, antiviral, antirheumatic, anticancer, and neuroprotective effects. However, curcumin is limited in terms of water-solubility, and curcumin analogues as well. Researchers have recently begun using biopolymers to encapsulate hydrophobic compounds in order to improve their bioavailability. The objective of this study was to evaluate the feasibility of using zein nanoparticles as an oral delivery vehicle for curcumin and its analogues. The investigation centered on the radical scavenging activities of ABTS and DPPH as well as the inhibition of α-glucosidase and xanthine oxidase by curcumin and its analogues, including CM-1, CM-2, CM-A, CM-F, and tetrahydrocurcumin. Fourier transform infrared spectroscopy was used to identify and characterize zein-curcumin analogues nanoparticles. All of the analogues except CM-1 presented ABTS radical scavenging activity on par with or superior to that of vitamin E. The α-glucosidase inhibitory activity of the compounds in this study was as follows: CM-F > CM > CM-A > CM-1. The α-glucosidase inhibitory effects of curcumin and its analogues were superior to those of quercetin. Furthermore, only CM-F had a good xanthine oxidase inhibitory activity (IC50 = 2.05 ± 0.15 μM), suggesting a better inhibition than that of allopurinol (IC50 = 12.04 ± 1.54 μM). Amide I, amide II and amide III of zein were observed in the nanoparticles with the peak positions either existed or slightly shifted. The main peaks of all compounds themselves were also occurred or shifted in nanoparticles, indicating that partial hydrogen bonds occurred between the hydrophobic region of zein and compounds. The resulting nanoparticles without surfactant were spherical, small (mean particle size ≈ 125-150 nm), and had a narrow size distribution (polydispersity index < 0.4). Additionally, the encapsulated compounds were in an amorphous as detected by differential scanning calorimetry. These results demonstrate that zein can indeed be used as a carrier of curcumin and its analogues. Especially, zein-CM-F nanoparticle is the most potential inhibitors using in the treatment of gout and diabetes.

I. Experimental motivation and objective..........................................1
II. Literature review.............................................................3
A. Zein...........................................................................3
B. Preparation of zein nanoparticles..............................................6
D. α-Glucosidase inhibitors.......................................................6
E. Xanthine oxidase...............................................................8
F. Xanthine oxidase inhibitors....................................................9
G. Curcumin......................................................................11
H. Curcumin analogues............................................................11
III. Material and Methods........................................................14
A. Experimental design...........................................................14
B. Curcumin and its analogues....................................................15
C. ABTS free radical scavenging assay............................................16
D. DPPH free radical scavenging assay............................................17
E. Inhibitory activity of α-glucosidase assay....................................18
F. Inhibitory activity of XO assay...............................................19
G. Preparation of compounds loaded zein nanoparticles............................20
H. Particle size and zeta potential measurements.................................20
I. Particle yield and encapsulation efficiency...................................21
J. Fourier transform infrared (FTIR) spectroscopy................................22
K. Differential scanning calorimetry (DSC).......................................22
L. Transmission electron microscopy (TEM)........................................23
M. Statistical analysis..........................................................23
IV. Results and discussion.......................................................24
Part 1: Characterization of curcumin and curcumin analogues
nanoparticles as inhibitors of α-glucosidase.....................................24
A. Inhibition of α-glucosidase...................................................24
B. Evaluation of ABTS antioxidant activity.......................................28
C. Physicochemical characterizations of nanoparticles............................31
D. DSC...........................................................................37
E. FTIR spectrometry.............................................................40
Part 2: Characterization of curcumin and curcumin analogues
nanoparticles as inhibitors of xanthine oxidase..................................44
A. Inhibition of xanthine oxidase................................................44
B. Evaluation of DPPH antioxidant activity.......................................48
C. Physicochemical characterizations of nanoparticle.............................51
D. DSC...........................................................................55
E. FTIR spectrometry.............................................................58
F. Mechanism of nanoparticles preparation........................................61
V. Conclusions...................................................................63
VI. References...................................................................64
VII. Supporting information......................................................72

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陳楚澤（民104）。薑黃素及其衍生物抑制黃嘌呤氧化酶和α-葡萄糖苷酶之研究（未出版之碩士論文）。天主教輔仁大學，新北市。