| 黄洁雯 宋珍 郭晓奎 李擎天.抗菌肽人beta防御素3 融合糖类结合域的真核表达和对耐甲氧西林
金黄色葡萄球菌N315 的抑菌作用[J].,2014,14(31):6006-6010 |
| 抗菌肽人beta防御素3 融合糖类结合域的真核表达和对耐甲氧西林
金黄色葡萄球菌N315 的抑菌作用 |
| The Eukaryotic Expression of Antimirobial Peptide Human beta Defensin 3 -Carbohydrate-binding Domain and the Inhibition on Methicillin Resistant Staphylococcus AureusN315 |
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| DOI： |
| 中文关键词: 抑菌作用 耐甲氧西林金黄色葡萄球菌 抗菌肽 真核表达 |
| 英文关键词: Inhibitory effect Methicillin resistant Staphylococcus aureus Antimicrobial peptide Eukaryotic expression |
| 基金项目:国家自然科学基金项目（81000708） |
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| 中文摘要: |
| 目的：研究重组真核抗菌肽人beta防御素3-糖类结合域(human beta defensin 3 - carbohydrate-binding domain，hBD3-CBD)对耐甲
氧西林金黄色葡萄球菌(Methicillin resistant Staphylococcus aureus，MRSA) N315 的抑菌作用。方法：用重叠延伸剪接技术将抗菌肽
和融合抗菌肽hBD3、hBD3-CBD1、hBD3-CBD2的基因序列克隆入真核表达载体pVAX1 并转染HEK293T细胞，通过逆转录聚合酶链反应(reverse transcription-polymerase chain reaction，RT-PCR)、Western blot 方法检测其表达情况；并通过感染后细菌计数和
上清液白细胞介素6(interleukin-6，IL-6)浓度来检测抗菌肽转染HEK293T 细胞对MRSA N315 的抑制作用。结果：抗菌肽hBD3、
hBD3-CBD1、hBD3-CBD2 转染HEK293T 细胞后均有表达；感染后6 和12 小时，hBD3、hBD3-CBD1 和hBD3-CBD2 均显示出对MRSA N315 的抑菌作用；感染后12 小时，hBD3-CBD1 和hBD3-CBD2 的抑菌作用优于hBD3，hBD3-CBD1 和hBD3-CBD2 之间
的抑菌作用没有差异；感染后6 和12 小时，hBD3、hBD3-CBD1、hBD3-CBD2 的细胞上清液IL-6 浓度显著低于对照组，而
hBD3-CBD1 和hBD3-CBD2 两组之间的IL-6 浓度未见差异。结论：真核表达抗菌肽hBD3、hBD3-CBD 对MRSA N315 对
HEK293T 细胞的感染有显著抑制作用，且重组抗菌肽hBD3-CBD 的直接抑菌作用优于hBD3。 |
| 英文摘要: |
| Objective:To analyze the inhibition effects of recombinant eukaryotic antimicrobial peptide human beta defensin 3 -
carbohydrate-binding domain (hBD3-CBD) against Methicillin resistant Staphylococcus aureus (MRSA) N315.Methods:Gene splicing
by overlap extension was used to combine antimicrobial peptide and fusion antimicrobial peptide gene hBD3, hBD3-CBD1 and
hBD3-CBD2 with eukaryotic vector pVAX1; the recombinant plasmids were transfected in HEK293T cells and reverse transcription
PCR and western blot were used to detect the plasmid expressions; bacterial counting and supernatant IL-6 concentration were analyzed
to show the inhibition on MRSA N315 from antimicrobial peptide transfected HEK293T cells.Results:Antimicrobial peptide hBD3,
hBD3-CBD1 and hBD3-CBD2 could be expressed in transfected HEK293T cells; hBD3, hBD3-CBD1 and hBD3-CBD2 showed the
bactericidal activities against MRSA N315 at 6 and 12 hours after the infection; the bactericidal activities in hBD3-CBD1 and
hBD3-CBD2 were stronger than that in hBD3; there was no difference in the bactericidal activities between hBD3-CBD1 and
hBD3-CBD2; the supernatant IL-6 concentration in hBD3, hBD3-CBD1 and hBD3-CBD2 were lower than that in the control group, and
there was no difference in the IL-6 concentration between hBD3-CBD1 and hBD3-CBD2.Conclusion:Eukaryotic expressed
antimicrobial peptides hBD3 and hBD3-CBD can significantly inhibit the infection on HEK293T cells by MRSA N315; the direct
inhibition effects of recombinant antimicrobial peptide hBD3-CBD were stronger than that of hBD3. |
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