蓮蓬萃取物應用於保肝護腎之研究

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题名:	蓮蓬萃取物應用於保肝護腎之研究 The study of application of lotus seedpod extracts on hepatic and renal protection
作者:	賴彥勳 Lai, Yen-Hsun
贡献者:	陳璟賢
关键词:	非酒精性脂肪肝疾病;慢性腎臟疾病;蓮蓬;氧化壓力;細胞凋亡;纖維化 non-alcoholic fatty liver disease;chronic kidney disease;lotus seedpod extracts;oxidative stress;cell apoptosis;fibrosis
日期:	2017
上传时间:	2018-03-20T07:31:59Z (UTC)
摘要:	蓮蓬(lotus seedpod)為傳統中藥材，研究顯示具有抗氧化、抗老化與抗發炎作用，本實驗室先前的研究顯示蓮蓬對於肝腎損傷可能具有改善效果，但是蓮蓬在保肝護腎作用上仍需進一步研究。在保肝細胞實驗中，透過油酸(oleic acid, OA)誘導人類肝細胞株HepG2細胞損傷，而蓮蓬萃取物(lotus seedpod extracts, LSE)與其主要功效成分表沒食子兒茶素(epigallocatechin, EGC)可以增加HepG2細胞存活率並減少細胞中脂肪堆積；此外oleic acid誘導氧化壓力與細胞凋亡的增加，在LSE或EGC的給予下，皆能降低氧化壓力與抑制細胞凋亡來減少肝細胞損傷，分子數據顯示LSE的抗凋亡作用可能通過線粒體途徑介導。在護腎動物實驗中，則透過10週連續餵食0.2% 腺嘌呤(adenine)誘導C57BL/6 小鼠產生腎損傷，並於第5週開始餵食1%與2% LSE直到第10週實驗結束。與誘導組相較之下，LSE能降低尿素氮(BUN)、肌酸酐(Creatinine, CRE)與發炎因子tumor necrosis factor-α (TNF-α)與interleukin 6 (IL-6)表現量，並且LSE可以增加腎臟總抗氧化能力與抗氧化酵素Glutathione peroxidase (GPx)、Glutathione reductase (GR)、catalase、Superoxide Dismutase (SOD) 活性並且減少脂質過氧化。另外LSE還能抑制腎臟纖維化及纖維化相關因子(TGF-β1、PAI-1、α-SMA、COL1A2)蛋白表現。研究結果顯示LSE具有潛力發展成為改善NAFLD與CKD的保健食品。Lotus seedpod, which is rich in flavonoid, is one of the traditional Chinese herbal medicines. Previous studies have shown that lotus seedpod possess antioxidant, anti-aging, and anti-inflammatory activities. However, the hepatic and renal protective effects of lotus seedpod are still unknown. In this study, we examined the heptoprotective effect of lotus seedpod extracts (LSE) and its main components epigallocatechin (EGC) in vitro. Firstly, oleic acid (OA) is used to induce the phenotype of non-alcoholic fatty liver disease (NAFLD) in human hepatic HepG2 cells. LSE dose-dependently improved the OA-induced loss of viability and lipid accumulation of HepG2 cells. Furthermore, LSE and EGC showed potential in reducing OA-induced occurrence of apoptosis confirmed by morphological and biochemical features, including an increase in oxidative stress, apoptotic bodies formation and caspase-3 activation. Molecular data showed the anti-apoptotic effect of LSE might be mediated via mitochondrial pathway. On the other hand, to investigate the protective effect and molecular mechanism of LSE on adenine-induced chronic kidney disease (CKD) mice in vivo, C57BL/6JNar1 mice were treated with 0.2% adenine, an inducer of chronic kidney injury (CKD) for 4 weeks, and then treated LSE (1% or 2%) for another 6 weeks. After 10 weeks, mice serum and kidney of mice were collected for further studies. The results showed that serum levels of blood urea nitrogen (BUN) and creatinine (CRE) in adenine-treated mice were increased as compared with control group. 1% and 2% LSE could decrease both levels when compared with the adenine alone group. In inflammatory cytokines analysis, we found that serum level of tumor necrosis factor (TNF)- α was significant increased after adenine induction, while 2% of LSE could reduce the serum level. In determination of oxidative stress in kidney, adenine could increase thiobarbituric acid relative substances (TBARS) level, while LSE treatments could significantly decrease oxidative stress level in mice. In measurement of antioxidant enzyme activity, adenine also decrease glutathione peroxidase (GPx) activity, glutathione (GSH) level, glutathione reductase (GR), catalase and superoxide dismutase (SOD) activity in kidney tissues, while LSE treatments could increase these enzyme activities. Furthermore, adenine-induced renal morphology fibrosis and fibrosis related proteins (TGF-β1, PAI-1, α-SMA, COL1A2) were decreased after treated with LSE. Our data imply that LSE potentially could be developed as an anti-NAFLD and anti-CKD agents in future.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/18852
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