亞抑菌濃度β內(nèi)酰胺類藥物對金黃色葡萄球菌α溶血素的表達調(diào)控作用
發(fā)布時間：2018-11-11 21:26
【摘要】：目的:探討亞抑菌濃度β內(nèi)酰胺類藥物對金黃色葡萄球菌(Staphylococcus aureus,S.aureus)致病物質(zhì)α溶血素(Alpha hemolysin,Hla)表達的調(diào)控作用及其調(diào)控機制。方法:①實驗金黃色葡萄球菌菌株分別收集于安徽醫(yī)科大學(xué)第一附屬醫(yī)院檢驗科培養(yǎng)臨床標(biāo)本,質(zhì)控菌株選用金黃色葡萄球菌ATCC29213;選擇青霉素、頭孢西丁、頭孢他啶、頭孢唑啉作為β內(nèi)酰胺類代表藥物,肉湯稀釋法分別測定實驗細菌對選用藥物的最小抑菌濃度(minimal inhibitory concentration,MIC);②實驗金黃色葡萄球菌菌株經(jīng)過培養(yǎng)8h后,分別收集細菌與培養(yǎng)上清。熒光定量PCR檢測細菌α溶血素mRNA的表達變化,免疫印跡法檢測細菌培養(yǎng)上清中α溶血素蛋白的表達變化,紅細胞溶血實驗檢測細菌培養(yǎng)上清中溶血活性的水平變化;③取金黃色葡萄球菌溶血活性表達較強的菌株,分別在培養(yǎng)的同時加入1/8MIC、1/4MIC、1/2MIC亞抑菌濃度的β內(nèi)酰胺類藥物,分為青霉素、頭孢西丁、頭孢他啶和頭孢唑啉作用組;平板菌落計數(shù)法計數(shù)繪制生長曲線,組設(shè)立4h、8h、12h和24h四個培養(yǎng)觀察時間點,分別檢測細菌與培養(yǎng)上清中α溶血素表達變化,以及細菌培養(yǎng)上清中溶血活性的水平變化;以未加入藥物培養(yǎng)的細菌作為正常對照。④熒光定量PCR檢測金黃色葡萄球菌轉(zhuǎn)錄調(diào)控因子AgrA在亞抑菌濃度β內(nèi)酰胺類藥物作用下的表達變化。結(jié)果:①不同金黃色葡萄球菌菌株之間α溶血素mRNA轉(zhuǎn)錄水平、上清中的蛋白翻譯水平的表達量以及培養(yǎng)上清的溶血活性存在差異;②1/8MIC、1/4MIC亞抑菌濃度β內(nèi)酰胺類藥物對金黃色葡萄球菌生長無明顯抑制作用,可顯著上調(diào)細菌α溶血素的表達;作用8h后,即可顯著上調(diào)α溶血素mRNA、上清中蛋白的表達,提高培養(yǎng)上清對紅細胞的溶血作用;③亞抑菌濃度β內(nèi)酰胺類藥物對金黃色葡萄球菌作用后,可顯著上調(diào)細菌轉(zhuǎn)錄調(diào)控因子AgrA的表達,結(jié)果顯示可通過AgrA上調(diào)α溶血素mRNA的表達。結(jié)論:亞抑菌濃度β內(nèi)酰胺類藥物能夠顯著上調(diào)金黃色葡萄球菌致病物質(zhì)α溶血素的表達,其機制可能通過細菌轉(zhuǎn)錄調(diào)控因子AgrA的表達調(diào)控。
[Abstract]:Aim: to investigate the regulatory effect of 尾 lactams on the expression of 偽 hemolysin (Alpha hemolysin,Hla) in Staphylococcus aureus (Staphylococcus aureus,S.aureus). Methods: 1 the strains of Staphylococcus aureus were collected from the laboratory of the first affiliated Hospital of Anhui Medical University, and Staphylococcus aureus ATCC29213; was used as the quality control strain. Penicillin, cefoxitin, ceftazidime and cefazolin were selected as representative drugs of 尾 -lactam. The minimal inhibitory concentration (minimal inhibitory concentration,MIC) of experimental bacteria against selected drugs was determined by broth dilution method. 2Staphylococcus aureus strains were collected and supernatant cultured for 8 h. The expression of 偽 -hemolysin mRNA was detected by fluorescence quantitative PCR, the expression of 偽 -hemolysin protein in the supernatant of bacterial culture was detected by Western blot, and the hemolytic activity in the supernatant of bacterial culture was detected by erythrocyte hemolysis test. 3 the strains with strong hemolytic activity of Staphylococcus aureus were cultured with 尾 lactams of 1 / 8 MIC1 / 4 MIC1 / 2 MIC and divided into penicillin and cefoxitin. Ceftazidime and cefazolin group; The growth curve was plotted by plate colony counting method. The four culture observation time points of 4 h, 8 h, 12 h and 24 h were set up to detect the changes of 偽 hemolysin expression in bacteria and culture supernatant, and the level of hemolytic activity in bacteria culture supernatant. The expression of staphylococcus aureus transcription regulator AgrA was detected by fluorescence quantitative PCR under the subinhibitory concentration of 尾 lactams. Results: 1 there were differences in the transcription level of 偽 -hemolysin mRNA, the level of protein translation in supernatant and the hemolytic activity of culture supernatant among different Staphylococcus aureus strains. The growth of Staphylococcus aureus was not inhibited by 尾 -lactams, but the expression of 偽 -hemolysin was significantly up-regulated. After 8 h treatment, the expression of protein in the supernatant of 偽 hemolysin mRNA, could be upregulated significantly, and the hemolytic effect of culture supernatant on erythrocyte was increased. (3) 尾 -lactam could significantly up-regulate the expression of AgrA, the expression of 偽 -hemolysin mRNA, after the treatment of staphylococcus aureus with subinhibitory concentration of 尾 -lactam. The results showed that the expression of 偽 -hemolysin mRNA could be upregulated by AgrA. Conclusion: 尾 lactams can significantly up-regulate the expression of 偽 hemolysin, a pathogenic substance of Staphylococcus aureus, and its mechanism may be regulated by the expression of bacterial transcription regulator AgrA.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.5
[Abstract]:Aim: to investigate the regulatory effect of 尾 lactams on the expression of 偽 hemolysin (Alpha hemolysin,Hla) in Staphylococcus aureus (Staphylococcus aureus,S.aureus). Methods: 1 the strains of Staphylococcus aureus were collected from the laboratory of the first affiliated Hospital of Anhui Medical University, and Staphylococcus aureus ATCC29213; was used as the quality control strain. Penicillin, cefoxitin, ceftazidime and cefazolin were selected as representative drugs of 尾 -lactam. The minimal inhibitory concentration (minimal inhibitory concentration,MIC) of experimental bacteria against selected drugs was determined by broth dilution method. 2Staphylococcus aureus strains were collected and supernatant cultured for 8 h. The expression of 偽 -hemolysin mRNA was detected by fluorescence quantitative PCR, the expression of 偽 -hemolysin protein in the supernatant of bacterial culture was detected by Western blot, and the hemolytic activity in the supernatant of bacterial culture was detected by erythrocyte hemolysis test. 3 the strains with strong hemolytic activity of Staphylococcus aureus were cultured with 尾 lactams of 1 / 8 MIC1 / 4 MIC1 / 2 MIC and divided into penicillin and cefoxitin. Ceftazidime and cefazolin group; The growth curve was plotted by plate colony counting method. The four culture observation time points of 4 h, 8 h, 12 h and 24 h were set up to detect the changes of 偽 hemolysin expression in bacteria and culture supernatant, and the level of hemolytic activity in bacteria culture supernatant. The expression of staphylococcus aureus transcription regulator AgrA was detected by fluorescence quantitative PCR under the subinhibitory concentration of 尾 lactams. Results: 1 there were differences in the transcription level of 偽 -hemolysin mRNA, the level of protein translation in supernatant and the hemolytic activity of culture supernatant among different Staphylococcus aureus strains. The growth of Staphylococcus aureus was not inhibited by 尾 -lactams, but the expression of 偽 -hemolysin was significantly up-regulated. After 8 h treatment, the expression of protein in the supernatant of 偽 hemolysin mRNA, could be upregulated significantly, and the hemolytic effect of culture supernatant on erythrocyte was increased. (3) 尾 -lactam could significantly up-regulate the expression of AgrA, the expression of 偽 -hemolysin mRNA, after the treatment of staphylococcus aureus with subinhibitory concentration of 尾 -lactam. The results showed that the expression of 偽 -hemolysin mRNA could be upregulated by AgrA. Conclusion: 尾 lactams can significantly up-regulate the expression of 偽 hemolysin, a pathogenic substance of Staphylococcus aureus, and its mechanism may be regulated by the expression of bacterial transcription regulator AgrA.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.5
【相似文獻】
相關(guān)期刊論文 前10條
1 白潔;機組配餐員的金黃色葡萄球菌污染情況[J];國外醫(yī)學(xué)(衛(wèi)生學(xué)分冊);2002年02期
2 朱自強;;金黃色葡萄球菌更具殺傷力的原因在于其金黃色“外殼”[J];基礎(chǔ)醫(yī)學(xué)與臨床;2005年11期
3 杜洪利;張雙雙;歐旭;閆訓(xùn)友;;金黃色葡萄球菌檢測技術(shù)研究進展[J];食品與發(fā)酵科技;2009年05期
4 黃嶺芳;賴衛(wèi)華;張莉莉;;食品中金黃色葡萄球菌快速檢測方法的研究進展[J];食品與機械;2009年06期
5 王娉;勞華均;胡s，
本文編號：2326183
本文鏈接：http://sikaile.net/huliyixuelunwen/2326183.html
最近更新
教材專著
