二氢丹参酮Ⅰ联合氨苄西林对生物膜阳性MRSE耐药基因表达的影响
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摘要
目的:探讨二氢丹参酮Ⅰ联合氨苄西林对生物膜阳性的耐甲氧西林表皮葡萄球菌(methicillin resistant S.epidermidis,MRSE)耐药基因mecA表达的影响。方法:筛选mecA阳性MRSE临床菌株,体外建立生物膜模型,使用抑制生物膜最小的联合用药部分抑菌浓度指数(fractional inhibitory concentration index,FICI)药物组合、各单用药物以及阳性对照药物万古霉素进行干预,qRT-PCR检测mecA基因、ELISA检测青霉素结合蛋白2a(penicilin binding protein 2a,PBP2a)的表达。结果:2μg﹒mL~(-1)氨苄西林与2μg﹒mL~(-1)二氢丹参酮Ⅰ联用组抑制了SE21021菌株mecA、PBP2a的表达(P<0.05),且联用组PBP2a低于氨苄西林组、万古霉素组(P<0.05)。各组药物均促进了SE26508菌株mecA基因的表达(P<0.05),但是联用组mecA基因的表达相对低于氨苄西林单用组、二氢丹参酮Ⅰ单用组、万古霉素组(P<0.05);联用组能够抑制PBP2a的表达,但是效果不如氨苄西林单用组、二氢丹参酮Ⅰ单用组(P>0.05)。联用组同时抑制了SE30291菌株mecA和PBP2a蛋白的表达,但无统计学意义(P>0.05)。结论:二氢丹参酮Ⅰ和氨苄西林联用可能对生物膜阳性MRSE具有抑制作用,但需要加大菌株数量进一步深入研究求证。
Objective: To investigate the effect of dihydrotanshinone I combined with ampicillin on the expression of mecA gene of methicillin resistant S. epidermidis(MRSE) resistant to biofilm. Method: The mecA positive MRSE clinical isolates were screened and the biofilm model was established in vitro. The combination of fractional inhibitory concentration index(FICI) which was the smallest inhibitory concentration combination of biofilm was used. The drugs were used alone and the positive control drug was vancomycin. The mecA gene was detected by qRT-PCR and the expression of penicilin binding protein 2a(PBP2a) was detected by ELISA. Result: The expression of mecA and PBP2a in SE21021 strain was inhibited by 2 μg﹒mL~(-1) ampicillin and 2 μg﹒mL~(-1) dihydrotanshinone Ⅰ combination group(P<0.05). The PBP2a in combination group was lower than that of ampicillin group and vancomycin group(P<0.05). The expression of mecA gene in SE26508 strain was promoted(P<0.05), but the expression of mecA gene in the combination group was lower than that in ampicillin group and dihydrotanshinone Ⅰ group and vancomycin group(P<0.05). The combination group could inhibit the expression of PBP2a, but the effect was not as good as that of ampicillin group and dihydrodanshinone Ⅰ group(P>0.05). The expression of mecA and PBP2a protein in SE30291 strain was also inhibited by the combination group, but had no statistical significance(P>0.05). Conclusion: The combination of dihydrotanshinone Ⅰ and ampicillin may inhibit the biofilm-positive MRSE, but it needs to be further studied.
Objective: To investigate the effect of dihydrotanshinone I combined with ampicillin on the expression of mecA gene of methicillin resistant S. epidermidis(MRSE) resistant to biofilm. Method: The mecA positive MRSE clinical isolates were screened and the biofilm model was established in vitro. The combination of fractional inhibitory concentration index(FICI) which was the smallest inhibitory concentration combination of biofilm was used. The drugs were used alone and the positive control drug was vancomycin. The mecA gene was detected by qRT-PCR and the expression of penicilin binding protein 2a(PBP2a) was detected by ELISA. Result: The expression of mecA and PBP2a in SE21021 strain was inhibited by 2 μg﹒mL~(-1) ampicillin and 2 μg﹒mL~(-1) dihydrotanshinone Ⅰ combination group(P<0.05). The PBP2a in combination group was lower than that of ampicillin group and vancomycin group(P<0.05). The expression of mecA gene in SE26508 strain was promoted(P<0.05), but the expression of mecA gene in the combination group was lower than that in ampicillin group and dihydrotanshinone Ⅰ group and vancomycin group(P<0.05). The combination group could inhibit the expression of PBP2a, but the effect was not as good as that of ampicillin group and dihydrodanshinone Ⅰ group(P>0.05). The expression of mecA and PBP2a protein in SE30291 strain was also inhibited by the combination group, but had no statistical significance(P>0.05). Conclusion: The combination of dihydrotanshinone Ⅰ and ampicillin may inhibit the biofilm-positive MRSE, but it needs to be further studied.
引文
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[5]李燕,李冬冬,陶传敏,等.表皮葡萄球菌生物膜形成及相关基因的检测及评价[J].中华医院感染学杂志.2010,20(4):473.
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[8]Carolina S,Ee Leen P,Kuan-Hon L,et al.Inhibition of penicillin-binding protein 2a(PBP2a)in methicillin resistant Staphylococcusaureus(MRSA)by combination of ampicillin and abioactive fraction from Duabanga grandiflora[J].BMC Complementary and Alternative Medicine,2015,15:178.
[9]段德军.丹参酮与万古霉素联合应用对MRSA的抑制作用[J].实用全科医学,2008,6(4);363.
[10]陆克乔.滇重楼正丁醇提取物抗白念珠菌生物膜作用及机制研究[D].安徽:安徽中医药大学,2016.
[2]Henning Büttner,Dietrich Mack,Holger Rohde.Structural basis of Staphylococcus epidermidis biofilm formation:mechanisms and molecular interactions[J].Frontiers in Cellular and Infection Microbiology,2015,5:4.
[3]崔新洁,夏瑾,邵铁娟等.中药抗耐甲氧西林金黄色葡萄球菌生物膜研究进展[J].中国中医药信息杂志.2017,24(2):132.
[4]李燕,李冬冬,宋増,等.4种方法检测表皮葡萄球菌生物膜形成能力的应用价值探讨[J].中国实验诊断学.2011,15(3):424.
[5]李燕,李冬冬,陶传敏,等.表皮葡萄球菌生物膜形成及相关基因的检测及评价[J].中华医院感染学杂志.2010,20(4):473.
[6]Lewis K.Riddle of biofilm resistance[J].Antimicrob Agents Chemother,2001,45(4):999.
[7]Patrick RG,Mitchell WP,Renee B,et al.Synergistic,collaterally sensitiveβ-lactam combinations suppress resistance in MRSA[J].Nature Chemical Biology,2015,11:855.
[8]Carolina S,Ee Leen P,Kuan-Hon L,et al.Inhibition of penicillin-binding protein 2a(PBP2a)in methicillin resistant Staphylococcusaureus(MRSA)by combination of ampicillin and abioactive fraction from Duabanga grandiflora[J].BMC Complementary and Alternative Medicine,2015,15:178.
[9]段德军.丹参酮与万古霉素联合应用对MRSA的抑制作用[J].实用全科医学,2008,6(4);363.
[10]陆克乔.滇重楼正丁醇提取物抗白念珠菌生物膜作用及机制研究[D].安徽:安徽中医药大学,2016.