以表皮葡萄球菌LuxS为靶点的反义锁核酸对生物被膜的影响

发布时间：2018-06-08 22:20

本文选题：反义锁核酸 + 表皮葡萄球菌　；参考：《农业生物技术学报》2017年02期

【摘要】：细菌生物被膜的形成是导致细菌耐药性升高的主要原因,表皮葡萄球菌(Staphylococcus epidermidis)作为条件致病菌,一旦形成生物被膜后会粘附在医疗器械和病灶上造成反复感染,难以治疗。以细菌群体感应系统作为抑制生物被膜形成的研究靶点近年来受到广泛关注。S-核糖基高半胱氨酸酶(S-ribosyl homocysteine lyase,Lux S)是合成自诱导信号分子2(autoinducer-2,AI-2)的关键酶,构成了细菌内依赖Lux S基因产物的AI-2群体感应系统(AI-2/Lux S)。本研究以生物被膜形成初期为时间点,利用反义锁核酸技术阻断表皮葡萄球菌内AI-2/Lux S群体感应系统中的关键酶Lux S基因的表达,比较该基因被阻断前后对其他相关产膜基因的表达、Al-2信号分子的活性以及生物被膜形成能力等方面的影响。结果表明,针对Lux S m RNA序列设计的两条反义锁核酸序列LNA81和LNA352,相比而言,LNA81的抑制效果更佳,其他与被膜形成的相关基因除σ因子B(sigma factor B,sig B)和细胞间粘附因子AB(intercellular adhesion AB,ica AB)表达量上调外,附属调节子A(staphylococal accessory regulator A,sar A)、聚集相关蛋白(accumulation-associated protein,aap)、纤维蛋白原结合蛋白(fibrinogen-binding protein,fbe)和自溶素E(autolysin E,atl E)的表达量均下调,AI-2信号分子活性在一定时间内与对照组相比有所降低。由此说明,Lux S基因表达被部分抑制后会影响AI-2信号分子的活性以及其他被膜相关基因的表达,最终导致生物被膜形成能力减弱。本研究为深入探讨AI-2/Lux S群体感应系统在表皮葡萄球菌生物被膜形成过程中所起的调控作用提供了理论依据,同时也为抗产膜型表皮葡萄球菌形成的感染提供更多有效的潜在靶点和治疗方案。
[Abstract]:The formation of bacterial biofilm is the main reason leading to the increase of bacterial drug resistance. Staphylococcus epidermidisis, as a conditional pathogen, once formed, it will adhere to medical instruments and lesions and cause repeated infection, which is difficult to treat. In recent years, Sribosyl homocysteine lyase (Sribosyl homocysteine lyase) is a key enzyme in the synthesis of self-induced signaling molecule 2autoinducer 2 (AI-2) by using bacterial population sensing system as the target of inhibition of biofilm formation. The AI-2 population sensing system, which is dependent on Lux S gene products in bacteria, is AI-2 / Lux Sch. In this study, antisense locking nucleic acid (ASN) technique was used to block the expression of the key enzyme Lux S gene in AI-2 / Lux S population sensing system of Staphylococcus epidermidis at the early stage of biofilm formation. The effects of this gene on the expression of Al-2 signal molecule and the ability of biofilm formation were compared before and after blocking. The results showed that LNA81 and LNA352, two antisense lock-nucleotide sequences designed for Lux S m RNA sequence, were more effective than LNA81 in inhibiting LNA81. Other genes related to membrane formation were up-regulated except 蟽 factor sigma factor sig B) and intercellular adhesion ABAbs. In addition, the expression of A(staphylococal accessory regulator Acansar, aggregation associated protein, fibrinogen-binding protein (fibrinogen-binding protein), and self-lysin Eautolysin Etoatl (E) down-regulated the activity of AI-2 signal molecules in a certain period of time compared with that of the control group. These results suggest that the partial inhibition of Lux S gene expression may affect the activity of AI-2 signaling molecules and the expression of other membrane related genes, resulting in a decrease in the ability of biofilm formation. This study provides a theoretical basis for further study of the regulatory role of AI-2 / Lux S population sensing system in the biofilm formation of Staphylococcus epidermidis. At the same time, it also provides more effective potential targets and treatment for the infection caused by Membranous Staphylococcus epidermidis.
【作者单位】：宁夏大学西部特色生物资源保护与利用教育部重点实验室;宁夏大学生命科学学院;
【基金】：宁夏自然科学基金(No.NZ15019)
【分类号】：R378.11

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