山苦瓜萃取物對 β-類澱粉胜?誘導 PC12 細胞損傷之神經保護影響

ASIAIR > College of Medical and Health Science > Department of Medical Laboratory Science and Biotechnology > Theses & dissertations > Item 310904400/113649

Please use this identifier to cite or link to this item: http://asiair.asia.edu.tw/ir/handle/310904400/113649

Title:	山苦瓜萃取物對 β-類澱粉胜?誘導 PC12 細胞損傷之神經保護影響 Neuroprotective effects of wild bitter gourd extract on β-amyloid peptide-induced PC12 cell injury
Authors:	詹鎧毓 JHAN, KAI-YU
Contributors:	醫學檢驗暨生物技術學系
Keywords:	神經退化性疾病;抗神經發炎;山苦瓜;β-類澱粉胜? Neurodegenerative disease;Anti-neuroinflammation;Wild bitter gourd;β-amyloid peptide
Date:	2022-08-01
Issue Date:	2022-10-31 03:03:50 (UTC+0)
Publisher:	亞洲大學
Abstract:	阿茲海默症為一常見的神經退化性疾病，常發生於65歲以上的老年人，導致患者記憶喪失及認知衰退，該病症是由於腦部 -類澱粉胜?(-amyloid peptide, A) 斑塊的堆積，造成氧化壓力、粒線體失調、突觸萎縮，進而導致腦部功能退化。山苦瓜 (Momordica charantia Linn. var. abbreviate Ser.) 為葫蘆科苦瓜屬一?生蔓性攀緣草本植物，別名野苦瓜、短果苦瓜或小苦瓜，原產於熱帶亞洲地區。本研究利用山苦瓜作為實驗樣品，將山苦瓜果實含籽冷凍乾燥後磨碎成粉末後浸泡於 95% 乙醇進行萃取，經減壓濃縮去除乙醇後以不同極性之溶劑進行分配萃取，得到正己烷層 (BGH)、乙酸乙酯層 (BGE) 、正丁醇層 (BGB) 、及水層 (BGW) 四個分配萃取區分物。分別進行總酚含量測定及DPPH自由基清除能力試驗，並以 LPS 誘發小鼠巨噬細胞 (RAW264.7)探討其對發炎反應之影響；以 A1-40 誘發大鼠嗜鉻瘤細胞 (PC12)，探討其對神經損傷之預防及治療功效。結果發現BGE之總酚含量為1945 mg/100 g dry wt，對應其DPPH自由基清除能力達到53.4 %，具有良好的抗氧化能力。抑制發炎介質亞硝酸鹽生成試驗結果發現BGE在濃度200 ?g/mL時有67 %的抑制效果，具有顯著之抗發炎能力。於 RAW264.7細胞之蛋白質表現試驗結果發現，山苦瓜萃取物經由抑制 iNOS 蛋白質之表現，達到抗發炎效果；於PC12細胞之神經保護試驗發現山苦瓜正己烷及乙酸乙酯萃取物具有優秀預防及治療能力，於 PC12 細胞之蛋白質表現試驗結果發現，山苦瓜乙酸乙酯萃取物經由抑制 Bax及增加 Bcl-2 和HO-1 之表現量，達到神經保護及抗細胞凋亡效果。山苦瓜萃取物抑制乙醯膽鹼酯?增加腦部乙醯膽鹼的含量，達到神經保護效果。由本研究結果得知山苦瓜萃取物具有抗氧化、改善神經發炎、抗細胞凋亡及保護神經細胞之保健功效。未來可應用於阿茲海默症等神經退化性疾病保健食品之開發。 Alzheimer's disease is a common neurodegenerative disease that usually occurs in the elderly over 65 years old, causing memory loss and cognitive decline in patients. The disease is due to the accumulation of β-amyloid peptide (Aβ) plaques in the brain. Causes oxidative stress, mitochondrial dysregulation, synaptic atrophy, which in turn leads to degeneration of brain function. Momordica charantia Linn. var. abbreviate Ser. is an annual climbing herb of the bitter gourd family, short-fruited bitter gourd or small bitter gourd, native to tropical Asia. In this study, wild bitter gourd was used as an experimental sample. The wild bitter gourd whole fruit contains seeds were freeze-dried, then ground into powder, and then immersed in 95% ethanol for extraction. The ethanol was removed by concentration under reduced pressure, and then partitioned and extracted with solvents of different polarities to obtain n-hexane layer (BGH), ethyl acetate layer (BGE), n-butanol layer (BGB), and water layer (BGW) four partition extracts. Determination of total phenolic content and DPPH free radical scavenging ability test were performed respectively, and LPS-stimulated RAW264.7 macrophages were used to investigate their effects on inflammatory response. PC12 cells were induced by Aβ1-40, and its preventive and therapeutic effects on neurotoxicity were investigated. The results showed that the total phenolic content of BGE was 1945 mg/100 g dry wt, corresponding to DPPH free radical scavenging ability reaching 53.4 %, showing good antioxidant capacity. The results of the inhibition of nitrite generation test showed that BGE has a 14.8% inhibitory effect at a concentration of 100 ?g/mL, and has a significant anti-inflammatory ability. In the mouse macrophage inflammatory protein expression test, the anti-inflammatory effect was achieved by inhibiting the expression of iNOS; In the neuroprotection test of PC12 cells, it was found that the n-hexane and ethyl acetate extracts of balsam pear had excellent preventive and therapeutic abilities. The results of the protein expression test of PC12 cells found that the ethyl acetate extract of balsam pear inhibited Bax and increased Bcl-2. and HO-1 expression to achieve neuroprotective and anti-apoptotic effects. Bitter gourd extract inhibits acetylcholinesterase and increases the content of acetylcholine in the brain, achieving neuroprotective effect. From the results of this study, it is known that the extract of balsam pear has anti-oxidation, improving nerve inflammation, anti-apoptosis and protecting nerve cells. In the future, it can be applied to the development of health food for neurodegenerative diseases such as Alzheimer's disease.
Appears in Collections:	[Department of Medical Laboratory Science and Biotechnology] Theses & dissertations

Files in This Item:

File	Description	Size	Format
index.html		0Kb	HTML	83	View/Open

Loading...