- 學位論文
注射Propacetamol建構急性乙醯胺基苯酚肝毒性小鼠並探討高良薑萃取物與高良薑素的防毒性
Establish an Acute Acetaminophen Hepatotoxicity in Mice by Injection of Propacetamol and Study the Prevention of Hepatotoxicity by Alpinia officinarum Extracts and Galangin
摘要
根據衛生福利部2013年統計資料顯示，慢性肝病及肝硬化占2012年全國十大死亡原因第九名，此外肝癌為同年國人癌症十大死亡率的第二名，因此肝臟保健及有效治療藥物特別受到注目。之前實驗中，我們利用脂多醣誘導RAW264.7小鼠巨噬細胞做為發炎的細胞模式，發現高良薑萃取物（Alpinia officinarum extracts, AOE）與其主要成分－高良薑素（Galangin）具有顯著地抑制發炎能力。我們先嘗試以注射Propacetamol來建構急性乙醯胺基苯酚肝毒性（N-acetyl-p-aminophenol (APAP)-induced liver injury, AILI）模式小鼠，並探討高良薑萃取物與高良薑素的防毒性。乙醯胺基苯酚（Acetaminophen, APAP）為常見解熱與鎮痛劑的主要成份，當食用過量易產生肝、腎損傷，是歐、美最常見的化學性肝毒性之一。本實驗藉由單一次腹腔注射Propacetamol後，測試在不同誘導時間下小鼠血液中的麩丙酮酸轉胺酶（ALT）、麩草醋酸轉胺酶（AST）酵素活性，與肝臟切片染色結果，確定可藉由注射Propacetamol來產生AILI小鼠，並發現誘導第18小時為最嚴重肝損傷時間點。之後藉由誘導AILI小鼠前一小時分別管餵AOE、高良薑素、具有保肝功效的水飛薊素（Silymarin）與臨床急救APAP肝損傷之半胱胺酸醋酸（N-acetylcysteine, NAC）藥物，比較對AILI小鼠的保護效果，發現水飛薊素與NAC確實可對注射Propacetamol所誘導之AILI小鼠產生肝臟保護效果，此結果亦可證明透過注射Propacetamol所產生AILI小鼠來作為護肝藥物篩選實驗動物平台的可行性。此外，我們也證明AOE與高良薑素前處理，可產生顯著肝臟保護效果。以對小鼠注射雙倍劑量Propacetamol模擬臨床APAP致死劑量中毒病患，來誘導致死性肝損傷小鼠，並觀察比較注射後半小時餵食水飛薊素、NAC、AOE和高良薑素的小鼠存活情形，發現只有NAC能大幅提高小鼠存活時間與存活率，其他皆無急救效果。除此之外，臨床上APAP過量也可能造成急性腎臟損傷，我們藉由注射Propacetamol也來產生急性腎臟損傷模式（AIKI）的小鼠，並預處理水飛薊素、NAC與高良薑素，發現水飛薊素與高良薑素對此AIKI小鼠顯示保護效果。本實驗將現行由APAP誘導化學性肝、腎損傷動物模式進行改良，做為新的AILI與AIKI動物模式，並證明高良薑粗萃物與高良薑素可有效保護此模式動物的肝臟和腎臟。
並列摘要
According to Taiwan's 2013 statistics, chronic hepatitis and cirrhosis accounted for the ninth causes of death in 2012. Furthermore, hepatocellular carcinoma is the second most cancer cause of death in 2012. Therefore, clinical drugs or healthy foods that can cure or protect liver damages are an important research aim. In our previous study, we used lipopolysaccharide-induced mouse RAW264.7 macrophage cell line as an inflammatory cell model to screen which nature plants or Chinese herbals have anti-inflammatory activities. In these tested Chinese herbals, we found that Alpinia officinarum extract (AOE) and the major component of this extract, Galangin, have significant anti-inflammatory activities. In this study, we first established a novel acute N-acetyl-p-aminophenol (acetaminophen, APAP)-induced liver injury (AILI) model mice by single injection of propacetamol, a water-soluble precursor of APAP, and then study the protection effects of AOE and Galangin. APAP is a widely used counter analgesic and antipyretic drug. However, at higher doses, APAP causes liver and/or kidney damages. The AILI is the most frequent chemical cause of liver injury in both American and European. We examined the time- and dosage-dependent effects of AILI by propacetamol injection in mice through analyses of ALT and AST activities. From these results, we conclude a novel AILI model mice can be established by single propacetamol injection, and found that the most severe time for liver injury is the 18 hours after injection. Furthermore, we compared the therapeutic effects of AOE and Galangin with Silymarin and N-acetylcysteine (NAC) in these AILI model mice. We found that both Silymarin and NAC can protect AILI in mice before propacetamol injection. Our results showed a novel AILI mouse model by propacetamol injection and these model mice are feasible for screening liver protective drugs. In addition, we also proved that both AOE and Galangin pre-treatment have significant protective effects of AILI mice. However, we observed that only NAC can greatly improve survival time and rate when a lethal dosage of propacetamol injection. Otherwise, we also observed APAP-induced kidney injury (AIKI) in these mice with propacetamol injection. Silymarin, AOE and Galangin showed kidney protection effects of these AIKI mice. Results established a new AILI and AIKI mouse models by single propacetamol injection. Using these model mice, we showed that AOE and Galangin have significant protective effects on liver and kidney injuries.