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研究生: 田安然
Tien, An-Jan
論文名稱: 抗發炎中草藥之開發應用與機轉
The Development, Application, and Mechanism of Anti-inflammatory Traditional Herbal Medicine
指導教授: 鄭劍廷
Chien, Chiang-Ting
學位類別: 博士
Doctor
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2018
畢業學年度: 106
語文別: 英文
論文頁數: 70
中文關鍵詞: Antrodia cinnamomeaHepatic inflammationFibrosisHepatocarcinogenesisReactive oxygen speciesAntcin KN-nitrosodiethylaminearterial thrombosisICAM-1monacolin KMonascus Adlayplatelet
英文關鍵詞: Antrodia cinnamomea, Hepatic inflammation, Fibrosis, Hepatocarcinogenesis, Reactive oxygen species, Antcin K, N-nitrosodiethylamine, arterial thrombosis, ICAM-1, monacolin K, Monascus Adlay, platelet
DOI URL: http://doi.org/10.6345/DIS.NTNU.SLS.027.2018.D01
論文種類: 學術論文
相關次數: 點閱:112下載:15
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  • 無中文摘要

    Traditional Chinese herbal medicine has been widely used for thousands of years for health promotion, disease prevention and treatment. the clinical effect of Traditional Chinese herbal medicine is remarkable, but the mechanism of those herbal medicine are incompletely clear. In our study, we chose two famous and widely used herbal medicine to explore the mechanism by determine the anti-inflammatory and anti-oxidant effect. We chose Antrodia cinnamomea and Monascus Adlay as the study subject.
    Antrodia cinnamomea (A. cinnamomea), a popular medicinal mushroom in Taiwan, is widely used for prevention or treatment of liver diseases. Systematic studies on the anti-inflammatory effect of A. cinnamomea and its molecular mechanisms have not yet been fully investigated. HPLC fingerprint analysis identified seven ergostane-type triterpenoids from A. cinnamomea water extract (ACW) including highest content of Antcin K (AC), Antcin C, Antcin H, Dehydrosulphurenic acid, Antcin B, Antcin A and Dehydroeburicoic acid. Here, we explored the effects and mechanisms of ACW and the highest content AC on N-nitrosodiethylamine (DEN) induced liver inflammation, fibrosis and carcinogenesis in rats. In in vitro study, we found ACW and AC dose-dependently scavenged O2-., H2O2 and HOCl amount by a chemiluminescence analyzer. In in vivo experiment, oral intake ACW and AC significantly inhibited DEN-enhanced hepatocellular inflammation, fibrosis and carcinoma by pathologic observation, the elevated bile and liver reactive oxygen species (ROS) amounts, plasma γ-glutamyl transpeptidase, and oxidative stress including 3-nitrotyrosine, 4-hydroxynonenal and Kuppfer cell infiltration (ED-1 stains) in the inflammatory livers. DEN enhanced nuclear factor-B (NF-B) translocation, whereas ACW and AC suppressed DEN-enhanced NF-B translocation through the inhibition of its upstream signaling of p85/phosphoinositide-3-kinase, mitogen activated protein kinase and CYP2E1 expression. In conclusion, DEN can induce hepatocellular inflammation, fibrosis and carcinoma through the increase in NF-B translocation to nucleus and oxidative injury. ACW and its active component, Antcin K, counteract DEN-induced hepatic injury and inflammation by the protective and therapeutic mechanisms of a direct scavenging ROS activity and an upregulation of antioxidant defense mechanisms.
     Monascus Adlay (MA) prepared from fungal fermentation of Monascus purpureus inoculating with cooked adlay contains high content of monakolin K (MK) and phenolic compounds. We explored whether MA and MK improve FeCl3-induced arterial thrombosis in rats. The rats were divided into control, FeCl3-treated rat carotid artery occlusion (TTO), TTO determined with one-week MA, and TTO determined with one-week MK. We compared MA or MK effects on oxidative stress by chemiluminescence amplification and immunohistochemistry, TTO by a transonic system, NFB, ICAM-1, endoplasmic reticulum stress CHOP and Nrf2 signaling by western blotting. MA or MK efficiently depressed O2-, H2O2 and HOCl levels, platelet activation and aggregation and H2O2-enhanced ICAM-1 and VCAM-1 expression in the endothelial cells. FeCl3 significantly increased NFB p65, 3-nitrotyrosine, CHOP and ICAM-1 expression, and decreased nuclear Nrf2 translocation and induces arterial thrombus formation. MA or MK pretreatment significantly elongated the level of FeCl3-induced TTO compared to TTO group, significantly decreased proinflammatory NF-B/ICAM-1 signaling, endoplasmic reticulum stress CHOP expression and decreased thrombotic area. MA or MK significantly preserved nuclear Nrf2 translocation. MA and MK exerted a similar protective effect in attenuating thrombus formation. We suggest MA is better than MK to improve FeCl3-induced arterial thrombosis.
    Through the study about Monascus Adlay (MA) and Antrodia cinnamomea, we found that anti-inflammatory and anti-oxidative effect are one of the main mechanism of Traditional herbal medicine. We can explore more about the mechanisms of the other traditional herbal medicine to discover the secret of Chinese Medicine.

    1. Abstract 3 2. Introduction 5 3. Materials and Methods 9 3.1 Chemicals 9 3.2 Experimental model of liver injury 10 3.3 In vivo and in vitro chemiluminescence recording for ROS activity 11 3.4 Inflammation, ED-1, 3-NT, 4-HNE, apoptosis and autophagy in the liver 13 3.5 Western Blotting for p85, MAPK and CYP2E1 in the livers 15 3.6 Preparation of Monascus adlay (MA) 16 3.7 Animals and MA Treatments 17 3.8 FeCl3-induced carotid arterial time to occlusion (TTO) 17 3.9 Grouping 18 3.10 Lucigenin-Enhanced Chemiluminescence (CL) Counts 18 3.11 Soluble form of Vascular ICAM-1 and ROS assay 19 3.12 In situ demonstration of ROS production and amount in the carotid artery 19 3.13 Immunoblot analysis for NFB, 3-NT, ICAM-1, CHOP, and Nrf2 20 3.14 Preparation of Platelet Suspensions for Platelet activation and aggregation 21 3.15 Expression of ICAM-1 and VCAM-1 on ROS-treated endothelial cells 22 3.16 Statistical analyses 23 4. Final Results 23 4.1 Ingredients of A. cinnamomea fruiting body 23 4.2 Scavenging O2-., H2O2 and HOCl ability by ACW and AC 24 4.3 Exploring Anti-inflammatory effect of AC and ACW by 3-NT and 4-HNE stain and ED-1 infiltration 24 4.4 Anti-fibrosis effect of AC and ACW 25 4.5 ACW or AC treatment inhibiting NF-κB mediated inflammation 25 4.6 inhibition of phosphorylated p85, phosphorylated MAPK and CYP2E1 expression by AC or ACW 26 4.7 Reducing DEN-evoked apoptosis and autophagy by AC and ACW in the livers 26 4.8 MA or MK pretreatment delays FeCl3-induced TTO 42 4.9 MA or MK reduces FeCl3-induced arterial thrombosis and endothelial 3-NT and ICAM-1 43 4.10 MK or MK inhibited platelet activation and aggregation and endothelial ICAM-1 and VCAM-1 expression 45 4.11 Scavenging O2-., H2O2 and HOCl ability by MA and MK 47 4.12 MA or MK treatment on FeCl3-enhanced arterial ROS and sICAM-1 levels 49 4.13 MA or MK on 3-NT, ICAM-1, CHOP and n-Nrf2 expression in FeCl3-treated arteries 51 5. Discussion 52 6. Conclusion 62 7. 未來展望: 63 8. References 64 9. 個人介紹 70

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