烏梅萃取物對臨床抗碳青黴烯克雷白氏肺炎桿菌之抗菌研究

Antibacterial Research on Smoked Plum Extract against Clinical Carbapenem Resistant Klebsiella pneumoniae

蔡榮櫸(Rong-Ju Tsai)

指導教授 ： 劉孝漢

義守大學/醫學科技學院/生物科技學系/碩士(2022年)

摘要

抗生素作為對抗細菌感染的有效治療方式已經運用了數十年，但近十餘年來因為抗生素濫用使得具有抗藥性之病原菌越來越多，世界衛生組織(WHO)也提出警告必須盡快找出新型抗生素。本研究以義大醫院提供2株臨床分離之抗藥性克雷白氏肺炎桿菌（Carbapenem Resistant Klebsiella pneumoniae，CRKP)用傳統中草藥－烏梅進行抗菌測試。我們根據CLSI使用MHB進行最小抑菌濃度(MIC)和最小殺菌濃度(MBC)的測試，並測試了毒殺動力曲線(Time-kill assay)，原本懷疑為低pH值導致細菌生長不良，所以用稀鹽酸調整pH值測試了細菌生長狀況。實驗結果表示烏梅對CRKP的MIC/MBC大約在14~28mg/ml，顯示烏梅萃取液具有抗微生物生長之活性，Time-kill assay結果則顯示烏梅在14mg/ml時可對EDAH-CRKP6產生抑制，28mg/ml則可達到99.9%抑制率，而在相近pH值的0.1N HCl滴定MH broth中在24小時後和烏梅對比兩者有明顯差異。和抗生素對比後發現抑制時間長度更優於臨床使用之最後一線抗生素。綜上所述，烏梅在體外測試時顯示出抗菌能力，有望成為治療及預防微生物感染和食物中毒的藥品或保健食品之潛力。

關鍵字

烏梅 ； 克雷白氏肺炎桿菌 ； 抗菌 ； 抗生素

並列摘要

Antibiotics have use decades in antibacterial infection treatment, but the amount and type of drug resistance pathogen increase for nearly ten years because abuse of antibiotics. The World Health Organization (WHO) also warned we need to find out new antibiotic as fast as possible before existing antibiotics failure.We wish to find out a new hope to antibacterial infection in traditional Chinese herbs. We according The Clinical & Laboratory Standards Institute (CLSI) method use Müller-Hinton broth (MHB) test minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). We also test Time-kill kinetics assays to record the bacterial growth. We doubt the inhibition of Smoked Plum is the low pH value cause bacterial poor growth, so we use dilute hydrochloric acid adjust the broth pH value to test the bacterial growth status. According result, the MIC/MBC of Smoked Plum resist CRKP about 14~28mg/ml. Showing antimicrobial growth activity of Smoked Plum extraction. The Time-kill assays show the Smoked Plum inhibition activity at 14mg/ml, when concentration increase to 28mg/ml can elimination to 99.9999% of pathogens on EDAH-CRKP6. Although the growth slowly in MH broth of similar pH value with 0.1N hydrochloric acid adjust. But after 24 hours, the hydrochloric acid adjust broth is failure, however the Smoked Plum still have activity to inhibit bacterial growth. When we take two kinds of antibiotics in clinical are the drug of last resort with the Smoked Plum extracts to try 24hrs Time killing assay. The result is shows the Smoked Plum extracts have better suppression effect than two kinds of antibiotics at 24hrs. In summary, the Smoked Plum show its antibacterial activity on in vitro test, and it use in traditional Chinese medical have thousands of years. It maybe can become a new hope of the drug or healthy food to therapy and safeguard the bacterial infection or food poisoning.

並列關鍵字

Smoked Plum ； Chinese herbs ； Klebsiella pneumoniae

參考文獻

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