The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temp...The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temperature-regulated control of the P_RP_L tandem promoters. As determined by SDS-PAGE and densitometry, the overexpressed protein comprised up to 18.5% and 10.9% of the total soluble protein in E. coli cells grown in shake flasks and in batch fermentation, respectively. The recombinant NAP-1/IL-8 was then purified to>95% homogeneity by gel filtration and cation exchange chromatography. The purified protein appeared as a single band on the SDS-PAGE gel and possessed potent chemotactic activity in the concentration of <10 ng/ml, as assayed by the agarose plate method. An early skin reactivity was also observed when the pure NAP-1/IL-8 was injected subcutaneously into the rabbits. The N-terminal 36 amino acid sequence of the recombinant NAP1/IL-8 was determined using the Edman method and was shown to be identical to that of the ntive protein.展开更多
目的观察大鼠哮喘内皮中性粒细胞激活肽-78(ENA-78),表达及布地奈德对其的影响,探讨中性粒细胞(NEU)参与哮喘发作的可能作用机制。方法采用大鼠哮喘模型,随机分成哮喘组、对照组、布地奈德组,流式细胞术测定血NEU ENA-78的表达。结果哮...目的观察大鼠哮喘内皮中性粒细胞激活肽-78(ENA-78),表达及布地奈德对其的影响,探讨中性粒细胞(NEU)参与哮喘发作的可能作用机制。方法采用大鼠哮喘模型,随机分成哮喘组、对照组、布地奈德组,流式细胞术测定血NEU ENA-78的表达。结果哮喘组(97.65±13.99)MFI NEU ENA-78的平均荧光强度显著高于对照组(50.79±8.66)MFI(P<0.01);布地奈德组(75.81±6.10)MFI NEU ENA-78的平均荧光强度显著性低于哮喘组,且高于对照组(均P<0.01),NEU ENA-78的表达水平与支气管肺泡灌洗液(BALF)细胞总数呈显著正相关(n=29,r=0.781,P<0.01)。结论哮喘大鼠NEU ENA-78的表达水平增加,NEU可能通过ENA-78参与哮喘发作的炎症过程;布地奈德可能部分通过下调NEU ENA-78,从而减轻哮喘气道炎症。展开更多
文摘The complete gene coding for human neutrophilactivating protein-1/interleukin-8 was synthesized using a semi-chemical semi-enzymatic method. The synthetic gene was then overexpressed in Escherichia coli under the temperature-regulated control of the P_RP_L tandem promoters. As determined by SDS-PAGE and densitometry, the overexpressed protein comprised up to 18.5% and 10.9% of the total soluble protein in E. coli cells grown in shake flasks and in batch fermentation, respectively. The recombinant NAP-1/IL-8 was then purified to>95% homogeneity by gel filtration and cation exchange chromatography. The purified protein appeared as a single band on the SDS-PAGE gel and possessed potent chemotactic activity in the concentration of <10 ng/ml, as assayed by the agarose plate method. An early skin reactivity was also observed when the pure NAP-1/IL-8 was injected subcutaneously into the rabbits. The N-terminal 36 amino acid sequence of the recombinant NAP1/IL-8 was determined using the Edman method and was shown to be identical to that of the ntive protein.
文摘目的观察大鼠哮喘内皮中性粒细胞激活肽-78(ENA-78),表达及布地奈德对其的影响,探讨中性粒细胞(NEU)参与哮喘发作的可能作用机制。方法采用大鼠哮喘模型,随机分成哮喘组、对照组、布地奈德组,流式细胞术测定血NEU ENA-78的表达。结果哮喘组(97.65±13.99)MFI NEU ENA-78的平均荧光强度显著高于对照组(50.79±8.66)MFI(P<0.01);布地奈德组(75.81±6.10)MFI NEU ENA-78的平均荧光强度显著性低于哮喘组,且高于对照组(均P<0.01),NEU ENA-78的表达水平与支气管肺泡灌洗液(BALF)细胞总数呈显著正相关(n=29,r=0.781,P<0.01)。结论哮喘大鼠NEU ENA-78的表达水平增加,NEU可能通过ENA-78参与哮喘发作的炎症过程;布地奈德可能部分通过下调NEU ENA-78,从而减轻哮喘气道炎症。
