蒲葵子/齊墩果酸萃取物修復大鼠肝纖維化之評估

肝臟疾病在世界上是一個很重要的健康問題。慢性肝損傷可導致纖維化和肝硬化，甚至可以發展為肝細胞癌。肝臟纖維化是肝臟受傷癒合過程中的一種自我保護機制。為了修補肝臟損傷，肝星狀細胞被活化並在損傷部位產生纖維化瘢痕。病毒感染、空氣污染與長期飲酒等因素會造成肝臟損傷；因此，肝星狀細胞啟動自我修復過程，導致疤痕組織積聚，最終導致肝纖維化。過去多半認為，肝纖維化是不可逆的。但現在有愈來愈多證據顯示，遠離傷肝因子與使用藥物治療，即使是重度肝纖維化，也有機會被「逆轉」。目前進行中治療肝纖維化的臨床試驗主要方向包括抑制肝細胞凋亡、減少氧化壓力、抑制肝星狀細胞活化、減少纖維化瘢痕形成和免疫調節。蒲葵子(Livistona chinensis)是盛行於民間的天然中藥，其應用範圍廣泛，可調整腸胃功能、促進食慾、改善便秘等日常問題。蒲葵子中含有黃酮、酚類、棕櫚酸等化合物，具有抗氧化作用、抗發炎反應、抗血管生成和抗腫瘤能力。本計畫第一部份主要研究蒲葵子萃取物的製備與體外細胞定性分析。透過不同濃度的蒲葵子萃取物處理在肝星狀細胞上，發現半最大抑制濃度(Half maximal inhibitory concentration，IC50)為0.4毫克每毫升(mg/mL)；在細胞型態與流式細胞儀分析也觀察到蒲葵萃取物造成的多為細胞晚期凋亡。在活性氧分析中，在濃度為 0.3毫克每毫升(mg/mL)時，蒲葵子萃取物顯著降低了白細胞介素6的產生，也代表著蒲葵子萃取物具有抗發炎能力。後續實驗會藉由大鼠肝纖維化模組(藉由硫代乙醯胺誘發肝纖維化)，來評估蒲葵子萃取物對於大鼠肝纖維化的修復效果。齊墩果酸(Oleanolic acid)是一種天然存在的五環三萜類化合物，常見於許多植物中。它的衍生物表現出多種生物活性，包括肝臟保護、抗發炎和抗氧化作用。過去許多文獻也指出空氣細顆粒物(fine particulate matter，PM2.5)會造成肝臟發炎與肝糖代謝異常；而過量飲酒會導致許多肝臟疾病的產生，例如：肝臟發炎、肝纖維化甚至造成肝硬化。本計畫的第二部份為建立空氣細顆粒物和酒精誘導的小鼠肝纖維化模型，試著模擬人類在日常生活中會接觸到的傷肝因子。接著使用脂質體齊墩果酸來治療，評估其抗發炎作用和肝纖維的改善程度。血液生化分析中，本研究發現脂質體齊墩果酸治療顯著降低了丙氨酸氨基轉移酶(alanine aminotransferase)、天冬氨酸氨基轉移酶(aspartate aminotransferase)和谷氨酰轉移酶(γ-glutamyltransferase)水平；在組織病理方面，脂質體齊墩果酸減輕了脂肪變性並改善了肝發炎指數(Ishak’s modified HAI score)。因此，脂質體齊墩果酸對空氣細顆粒物和酒精誘導的小鼠肝臟損傷具有抗發炎和修復作用。

關鍵字

肝纖維化；蒲葵子；水萃取物；齊墩果酸；脂質體；空氣細顆粒物

並列摘要

Liver diseases are very important health problems in the world. Chronic liver damage can lead to fibrosis and cirrhosis and can even develop into hepatocellular carcinoma. Liver fibrosis is a self-protective mechanism during liver wound healing. After liver injury, hepatic stellate cells are activated and create fibrotic scars at the injury site. Factors such as viral infection and chronic alcohol consumption and air pollution wound impair hepatic stellate cells. Therefore, hepatic stellate cells initiate a self-repair process, leading to the accumulation of scar tissue and ultimately liver fibrosis. In the past, it was mostly believed that liver fibrosis was irreversible. Liver fibrosis can be reversed by eliminating the etiologic agents or disrupting the pathogenic mechanisms of liver injury. Severe liver fibrosis might be reversible by some medication in recent literature reviews. Clinical trials in the treatment of hepatic fibrosis include inhibition of hepatocyte apoptosis, reduction of oxidative stress, inhibition of hepatic stellate cells activation, reduction of fibrotic scar evolution, and immune modulation. Livistona chinensis is a natural herb medicine. It has a wide range of daily applications, examples include improving gastrointestinal function, promoting appetite and constipation. Livistona chinensis contains flavonoids, phenols and palmitic acid, which have antioxidant effect, anti-inflammatory, anti-angiogenesis and anti-tumor abilities. First part of this project, we studied the preparation and in vitro qualitative analysis of Livistona chinensis extract. The half maximal inhibitory concentration (IC50) of Livistona chinensis extract for hepatic stellate cells was 0.4 mg/mL; cell morphology and flow cytometry revealed the late apoptosis effect of Livistona chinensis extract. Reactive oxygen species analysis revealed that Livistona chinensis extract significantly reduced the production of interleukin-6 at the concentration of 0.3 mg/mL, representing its anti-inflammatory properties. In the future work, the rats liver fibrosis model (thioacetamide-induced) will be used to evaluate the therapeutic effect of Livistona chinensis extract against liver fibrosis. Oleanolic acid is a naturally occurring pentacyclic triterpenoid and is widely distributed in plants. Oleanolic acid and its derivatives have several biological activities, including hepatoprotective, anti-inflammatory, and antioxidant effects. Many studies showed fine particulate matter (PM2.5) exposure wound cause nonalcoholic steatohepatitis and impair hepatic glucose metabolism. Excessive alcohol consumption can cause liver diseases, including liver inflammation, liver fibrosis and liver cirrhosis. In the second part of the project, we established the PM2.5 and alcohol-induced liver fibrosis in mice, trying to simulate people exposure to the two liver-damage factors in daily life. We treated with liposomal Oleanolic acid nanoparticles and evaluated the anti-inflammatory effect on exposed cells and the ameliorative effect against liver fibrosis. Liposomal Oleanolic acid nanoparticles treatment significantly reduced alanine aminotransferase, aspartate aminotransferase, and γ-glutamyltransferase levels; histologically, it alleviated steatosis and improved Ishak’s modified HAI score. In conclusion, liposomal Oleanolic acid nanoparticles have potential anti-inflammatory and reparative effects for PM2.5 and alcohol-induced liver injury.

並列關鍵字

Liver fibrosis ； Livistona chinensis ； water extract ； oleanolic acid ； liposome ； PM2.5

參考文獻

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