大黃對METHICILLIN抗藥性金黃色葡萄球菌之抗菌活性研究

The antibacterial activity of Rhei Rhizoma against methicillin-resistant Staphylococcus aureus

紀昆霖(Kun-Lin Chi)

指導教授 ： 楊玲玲 ； 梁文俐

臺北醫學大學/藥學院/生藥學研究所/碩士(2001年)

摘要

金黃色葡萄球菌 (Staphylococcus aureus) 是人類主要的致病原之一而且其常對多數之抗生素具有抗藥性，特別是MRSA (methicillin-resistant Staphylococcus aureus)。MRSA感染所引起的敗血症是造成燒傷、糖尿病、手術後、愛滋病或癌症住院病患的重要死因之一。萬古黴素 (vancomycin) 目前幾乎已成為全球所接受用於治療MRSA感染的首選藥物 (drug of choice)，然而由於vancomycin抗藥性的產生，致使新型抗生素之開發成為醫藥界的重要課題，本研究以瓊脂紙錠擴散法 (disc agar diffusion method) 檢測二十種中藥材對由皮膚膿瘍及傷口所分離出之金黃色葡萄球菌的抗菌活性，並期找到新的抗菌物質。結果發現大黃 (Rhei Rhizoma) 及黃芩 (Scutellariae Radix) 具有抗菌活性。大黃50% 酒精萃取物經Sephadex LH20 管柱層析後得到四個分離液，其中甲醇分離液具有較高的抗菌活性。經薄層分析後發現甲醇分離液中含有具抗菌活性之anthraquinone衍生物: 大黃素 (emodin) 及蘆薈大黃素 (aloe-emodin)。進一步探討中西藥物合併使用之活性，當emodin與aloe-emodin分別和oxacillin合併使用時，均能有效的抑制methicillin抗藥性金黃色葡萄球菌 (methicillin-resistant Staphylococcus aureus ATCC33591) 的生長，而產生球菌抗藥性之β-內醯胺環水解酵素 (β-lactamase) 的活性經由SDS-PAGE電泳法和gel renaturation 法測量後發現emodin具有抑制β-lactamase的作用。

關鍵字

methicillin抗藥性金黃色葡萄球菌 ； 大黃 ； 大黃素 ； 蘆薈大黃素 ； β-內醯胺環水解酵素 ； SDS-PAGE

並列摘要

Staphylococcus aureus is one of the major infectious pathogens of human and which is often resistant to a wide range of antibiotics, especially the MRSA (methicillin-resistant Staphylococcus aureus). MRSA induced sepsis is an important cause of mortality in hospitalized patients with burn injury, diabetes mellitus, post-operation, AIDS or tumors. Vancomycin is almost universally accepted as the drug of choice for the treatment of MRSA infections. However, because of the production of vancomycin-resistance, the development of new antibiotics has become an important work for medical and pharmaceutical industrial. In this study, the antibacterial activities of twenty kinds of Chinese medicine against Staphylococcus aureus isolated from skin lesions, abscesses and wounds were investigated by disc agar diffusion method expected to find new antibacterial materials. Rhei Rhizoma and Scutellariae Radix were found possessing antibacterial activities. Column chromatography of 50% ethanol extracts of Rhei Rhizoma on Sephadex LH-20 yielded four fractions and the methanol fraction performed higher antibacterial activity (MIC 256μg/ml). Emodin and aloe-emodin, anthraquinone derivatives with antibacterial activity, were found contained in methanol fraction by thin layer chromatography. When combined emodin or aloe-emodin with oxacillin, the growth of methicillin-resistant Staphylococcus aureus ATCC33591 was effectively inhibited. The production ofβ-lactamase which causes resistance to β-lactam antibiotics was estimated by SDS-PAGE and gel renaturation method and emodin were found possessing inhibitory activity toβ-lactamase production.

並列關鍵字

methicillin-resistant Staphylococcus aureus ； Rhei Rhizoma ； emodin ； aloe-emodin ； β-lactamase ； SDS-PAGE

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14.台北榮民總醫院醫院感染控制委員會八十六年度工作成果報告

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