α-螺旋抗菌肽與傳統(tǒng)抗生素的體內(nèi)和體外的協(xié)同抗菌作用研究
發(fā)布時間：2018-11-13 09:36
【摘要】：眾所周知，抗生素的泛濫使用導(dǎo)致耐藥性的菌株不斷出現(xiàn)，使得抗感染治療愈加的困難和復(fù)雜化。傳統(tǒng)的抗生素已經(jīng)無法滿足當(dāng)前的治療需求，而新型抗菌藥物的研發(fā)速率也跟不上病原微生物引起的感染性疾病的出現(xiàn)速率。因此，越來越多的人在關(guān)注新型抗菌藥物的研發(fā)，同時，也在積極探索新的治療手段。 抗菌肽（Antimicrobial peptides，，AMPs）是一類具有廣譜抗菌活性的小分子多肽。隨著研究的深入，人們發(fā)現(xiàn)抗菌肽在自然界中廣泛存在，包括病毒、細(xì)菌、兩棲類、昆蟲、高等動植物乃至人體。抗菌肽（AMPs）對革蘭陽性菌、革蘭陰性菌和真菌均有抑殺作用，很多抗菌肽還可以抗原蟲、抗病毒以及殺傷動物體內(nèi)的腫瘤細(xì)胞，是一類極具潛力的多肽類抗生素。但是，抗菌肽在體內(nèi)的不穩(wěn)定性以及較高的毒副作用極大地限制了抗菌肽的臨床應(yīng)用。雖然抗菌肽作為候選藥物應(yīng)用于臨床中還有一段距離，但是正在努力探索優(yōu)化條件。目前，人們在探索新型治療手段上已取得了一定的進(jìn)展�？股氐穆�(lián)合用藥指的是，兩種或者兩種以上的抗生素同時應(yīng)用于抗感染治療中。聯(lián)合用藥可以擴(kuò)大抗生素的抗菌范圍，還可以減緩細(xì)菌耐藥性的出現(xiàn)以及降低藥物毒性的作用。在先前的研究中，人們利用抗生素之間的聯(lián)合用藥成功地提高了抗生素的治療效力。但是，關(guān)于抗菌肽與傳統(tǒng)抗生素之間的聯(lián)合用藥的研究還相對較少。本研究旨在探索抗菌肽與傳統(tǒng)抗生素的聯(lián)合用藥，以期在提高其抗菌活性的同時降低多肽的毒副作用，并評估它們的療效，探究它們之間的協(xié)同作用機(jī)制。 首先，利用固相肽合成法，合成實(shí)驗(yàn)所需的6條多肽。同時，選取4種傳統(tǒng)抗生素（亞胺培南，頭孢吡肟，鹽酸作用氟沙星和萬古霉素），通過倍比稀釋法分別測定抗菌肽與抗生素的抗菌活性，評估其體外療效。數(shù)據(jù)表明，抗菌肽的最小抑菌濃度（Minimum inhibitory concentration, MIC）普遍小于傳統(tǒng)抗生素，說明抗菌肽擁有更好的抗菌效力，這與先前的研究一致。 其次，利用棋盤法測定抗菌肽與傳統(tǒng)抗生素聯(lián)合用藥的協(xié)同抗菌效力。實(shí)驗(yàn)中選取了3種革蘭氏陰性菌，3種革蘭氏陽性菌。結(jié)果表明，抗菌肽與傳統(tǒng)抗生素之間表現(xiàn)出較強(qiáng)的協(xié)同抗菌作用，沒有任何拮抗作用的出現(xiàn)。 最后，基于體外聯(lián)合用藥的結(jié)果，優(yōu)選抗菌肽PL-5和鹽酸左氧氟沙星作為實(shí)驗(yàn)用藥，進(jìn)行體內(nèi)動物試驗(yàn)。通過創(chuàng)建小鼠皮膚感染模型，測定PL-5與鹽酸左氧氟沙星之間的協(xié)同抗菌效力。研究結(jié)果顯示，抗菌肽與鹽酸左氧氟沙星表現(xiàn)出極強(qiáng)的體內(nèi)協(xié)同抗菌效力。考慮到二者不同的殺菌機(jī)制，我們認(rèn)為抗菌肽與傳統(tǒng)抗生素之間的存在互補(bǔ)性的協(xié)同抗菌作用機(jī)制，從而提高了抗菌效力。 通過該課題，我們可以確定，抗菌肽與傳統(tǒng)抗生素之間的聯(lián)合用藥可以作為一個有效的療法應(yīng)用于未來的抗菌感染治療中。
[Abstract]:It is well known that the prevalence of antibiotics leads to the emergence of drug-resistant strains, making anti-infection treatment more difficult and complicated. Traditional antibiotics can not meet the current treatment needs, and the rate of development of new antibiotics can not keep up with the emergence rate of infectious diseases caused by pathogenic microorganisms. Therefore, more and more people are paying attention to the research and development of new antimicrobial agents. Antimicrobial peptide (Antimicrobial peptides,AMPs) is a class of small peptides with broad spectrum antibacterial activity. With the development of research, antimicrobial peptides are widely found in nature, including viruses, bacteria, amphibians, insects, higher plants and animals and even human beings. Antimicrobial peptide (AMPs) can inhibit gram-positive bacteria, gram-negative bacteria and fungi. Many antimicrobial peptides can also inhibit antigens, antivirus and kill tumor cells in animals. However, the instability of antimicrobial peptides in vivo and high toxicity and side effects greatly limit the clinical application of antimicrobial peptides. Although antimicrobial peptides are still far from being used as candidates for clinical use, efforts are being made to explore optimal conditions. At present, people have made some progress in exploring new treatment methods. Combined use of antibiotics means that two or more antibiotics are used in anti-infective therapy at the same time. Combined use of antibiotics can expand the range of antibiotics, but also can reduce the emergence of bacterial drug resistance and reduce the toxicity of drugs. In previous studies, a combination of antibiotics was used to improve the efficacy of antibiotics. However, the research on the combination of antimicrobial peptides and traditional antibiotics is relatively rare. The purpose of this study was to explore the combination of antimicrobial peptides and traditional antibiotics in order to improve their antibacterial activity and reduce the toxic and side effects of peptides, evaluate their efficacy and explore the synergistic mechanism between them. Firstly, six polypeptides were synthesized by solid-phase peptide synthesis. Meanwhile, four traditional antibiotics (imipenem, cefepime, flufloxacin hydrochloric acid and vancomycin) were used to determine the antibacterial activity of antimicrobial peptides and antibiotics by double dilution method. The data showed that the minimum inhibitory concentration (Minimum inhibitory concentration, MIC) of antimicrobial peptides was generally lower than that of traditional antibiotics, which indicated that antimicrobial peptides had better antibacterial efficacy, which was consistent with previous studies. Secondly, chessboard method was used to determine the synergistic antibacterial efficacy of antimicrobial peptides combined with traditional antibiotics. Three Gram-negative bacteria and three Gram-positive bacteria were selected in the experiment. The results showed that there was a strong synergistic antibacterial effect between antimicrobial peptides and traditional antibiotics without any antagonism. Finally, the antibacterial peptide PL-5 and levofloxacin hydrochloride were selected as the experimental drugs. The synergistic antibacterial activity of PL-5 and levofloxacin hydrochloride was determined by establishing mouse skin infection model. The results showed that antimicrobial peptide and levofloxacin hydrochloride showed strong synergistic antibacterial effect in vivo. In view of their different bactericidal mechanisms, we think that there is a complementary synergistic antibacterial mechanism between antimicrobial peptides and traditional antibiotics, which improves the antibacterial efficacy. Through this study, we can confirm that the combination of antimicrobial peptides and traditional antibiotics can be used as an effective therapy for future antimicrobial infection treatment.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R965
本文編號：2328709
[Abstract]:It is well known that the prevalence of antibiotics leads to the emergence of drug-resistant strains, making anti-infection treatment more difficult and complicated. Traditional antibiotics can not meet the current treatment needs, and the rate of development of new antibiotics can not keep up with the emergence rate of infectious diseases caused by pathogenic microorganisms. Therefore, more and more people are paying attention to the research and development of new antimicrobial agents. Antimicrobial peptide (Antimicrobial peptides,AMPs) is a class of small peptides with broad spectrum antibacterial activity. With the development of research, antimicrobial peptides are widely found in nature, including viruses, bacteria, amphibians, insects, higher plants and animals and even human beings. Antimicrobial peptide (AMPs) can inhibit gram-positive bacteria, gram-negative bacteria and fungi. Many antimicrobial peptides can also inhibit antigens, antivirus and kill tumor cells in animals. However, the instability of antimicrobial peptides in vivo and high toxicity and side effects greatly limit the clinical application of antimicrobial peptides. Although antimicrobial peptides are still far from being used as candidates for clinical use, efforts are being made to explore optimal conditions. At present, people have made some progress in exploring new treatment methods. Combined use of antibiotics means that two or more antibiotics are used in anti-infective therapy at the same time. Combined use of antibiotics can expand the range of antibiotics, but also can reduce the emergence of bacterial drug resistance and reduce the toxicity of drugs. In previous studies, a combination of antibiotics was used to improve the efficacy of antibiotics. However, the research on the combination of antimicrobial peptides and traditional antibiotics is relatively rare. The purpose of this study was to explore the combination of antimicrobial peptides and traditional antibiotics in order to improve their antibacterial activity and reduce the toxic and side effects of peptides, evaluate their efficacy and explore the synergistic mechanism between them. Firstly, six polypeptides were synthesized by solid-phase peptide synthesis. Meanwhile, four traditional antibiotics (imipenem, cefepime, flufloxacin hydrochloric acid and vancomycin) were used to determine the antibacterial activity of antimicrobial peptides and antibiotics by double dilution method. The data showed that the minimum inhibitory concentration (Minimum inhibitory concentration, MIC) of antimicrobial peptides was generally lower than that of traditional antibiotics, which indicated that antimicrobial peptides had better antibacterial efficacy, which was consistent with previous studies. Secondly, chessboard method was used to determine the synergistic antibacterial efficacy of antimicrobial peptides combined with traditional antibiotics. Three Gram-negative bacteria and three Gram-positive bacteria were selected in the experiment. The results showed that there was a strong synergistic antibacterial effect between antimicrobial peptides and traditional antibiotics without any antagonism. Finally, the antibacterial peptide PL-5 and levofloxacin hydrochloride were selected as the experimental drugs. The synergistic antibacterial activity of PL-5 and levofloxacin hydrochloride was determined by establishing mouse skin infection model. The results showed that antimicrobial peptide and levofloxacin hydrochloride showed strong synergistic antibacterial effect in vivo. In view of their different bactericidal mechanisms, we think that there is a complementary synergistic antibacterial mechanism between antimicrobial peptides and traditional antibiotics, which improves the antibacterial efficacy. Through this study, we can confirm that the combination of antimicrobial peptides and traditional antibiotics can be used as an effective therapy for future antimicrobial infection treatment.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R965
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