中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2008年
2期
90-92
,共3页
高宏富%肖光夏%夏培元%任建东
高宏富%肖光夏%夏培元%任建東
고굉부%초광하%하배원%임건동
内毒素类%鲎抗内毒素因子%模拟肽
內毒素類%鱟抗內毒素因子%模擬肽
내독소류%후항내독소인자%모의태
Endotoxins%Limulus anti-lipopolysaccharide factor%Simulated peptide
目的 了解鲎抗内毒素因子模拟肽REMP2体外中和内毒素及抗菌活性的作用. 方法以多黏菌素B(PMB)为阳性对照,将REMP2、PMB与内毒素/脂多糖(LPS)溶液混合,使REMP2、PMB反应终浓度分别为100.00、10.00、1.00、0.10、0.01μmol/L,LPS为1 EU/mL.测定各溶液中LPS浓度,计算其中和率;以等渗盐水(NS)作为对照,分别将REMP2及PMB配制成10、20、40、80 μmol/L溶液,LPS配制成100μg/L,分别作用于LPS刺激的小鼠单核巨噬细胞,采用酶联免疫吸附测定法检测其肿瘤坏死因子α(TNF-α)含量;将REMP2加入大肠杆菌标准菌株菌液中,终浓度为40μmol/L,于加入后10、20、40 min在透射电镜下观察大肠杆菌的形态学变化. 结果浓度为0.10、10.00、100.00μmol/L的REMP2对LPS的中和率与阳性对照的PMB值相近(P>0.05);浓度为10、20、40、80μmol/L的REMP2作用于细胞后,其TNF-α含量[(1175-4-162)、(859-4-122)、(645±142)、(489±102)ng/L]均低于对照组[(3463±218)ng/L,P<0.01].REMP2作用后透射电镜下大肠杆菌内外膜变模糊、毛糙,菌体肿胀,胞质空泡变性. 结论 REMP2具有良好的中和LPs活性及杀菌作用.
目的 瞭解鱟抗內毒素因子模擬肽REMP2體外中和內毒素及抗菌活性的作用. 方法以多黏菌素B(PMB)為暘性對照,將REMP2、PMB與內毒素/脂多糖(LPS)溶液混閤,使REMP2、PMB反應終濃度分彆為100.00、10.00、1.00、0.10、0.01μmol/L,LPS為1 EU/mL.測定各溶液中LPS濃度,計算其中和率;以等滲鹽水(NS)作為對照,分彆將REMP2及PMB配製成10、20、40、80 μmol/L溶液,LPS配製成100μg/L,分彆作用于LPS刺激的小鼠單覈巨噬細胞,採用酶聯免疫吸附測定法檢測其腫瘤壞死因子α(TNF-α)含量;將REMP2加入大腸桿菌標準菌株菌液中,終濃度為40μmol/L,于加入後10、20、40 min在透射電鏡下觀察大腸桿菌的形態學變化. 結果濃度為0.10、10.00、100.00μmol/L的REMP2對LPS的中和率與暘性對照的PMB值相近(P>0.05);濃度為10、20、40、80μmol/L的REMP2作用于細胞後,其TNF-α含量[(1175-4-162)、(859-4-122)、(645±142)、(489±102)ng/L]均低于對照組[(3463±218)ng/L,P<0.01].REMP2作用後透射電鏡下大腸桿菌內外膜變模糊、毛糙,菌體腫脹,胞質空泡變性. 結論 REMP2具有良好的中和LPs活性及殺菌作用.
목적 료해후항내독소인자모의태REMP2체외중화내독소급항균활성적작용. 방법이다점균소B(PMB)위양성대조,장REMP2、PMB여내독소/지다당(LPS)용액혼합,사REMP2、PMB반응종농도분별위100.00、10.00、1.00、0.10、0.01μmol/L,LPS위1 EU/mL.측정각용액중LPS농도,계산기중화솔;이등삼염수(NS)작위대조,분별장REMP2급PMB배제성10、20、40、80 μmol/L용액,LPS배제성100μg/L,분별작용우LPS자격적소서단핵거서세포,채용매련면역흡부측정법검측기종류배사인자α(TNF-α)함량;장REMP2가입대장간균표준균주균액중,종농도위40μmol/L,우가입후10、20、40 min재투사전경하관찰대장간균적형태학변화. 결과농도위0.10、10.00、100.00μmol/L적REMP2대LPS적중화솔여양성대조적PMB치상근(P>0.05);농도위10、20、40、80μmol/L적REMP2작용우세포후,기TNF-α함량[(1175-4-162)、(859-4-122)、(645±142)、(489±102)ng/L]균저우대조조[(3463±218)ng/L,P<0.01].REMP2작용후투사전경하대장간균내외막변모호、모조,균체종창,포질공포변성. 결론 REMP2구유량호적중화LPs활성급살균작용.
Ohjective To investigate the role of REMP2 derived from limulus anti-lipopolysaccharide faetor in neutralizing endotoxin in vitro and its antibacterial activity. Methods (1)REMP2 and PMB in the concentrations of 100.00,10.00,1.00,0.10,0.01 μmol/L were respectively mixed with LPS (1 EU/mL),with PMB as positive control.The LPS concentrations in different specimens were determined by routine method,and the neutralizing percentage was respectively calculated.(2)After adding isotonic saline(NS),the final concentrations of REMP2 and PMB were 10,20,40,80 μmol/L,and the concentration of LPS was 100μg/L.The murine monoeytie macrophages were stimulated with LPS,then cuhured with REMP2 and PMB,with NS in culture as negative control.The content of tumor necrosis factor(TNF)-α was determined by ELlSA kit.(3)The morphologic changes of Escherichia coli.was observed under electron microscope at 10,20 and 40 minutes after addition of REMP2 to Escherichia coli suspension(with terminal concentration of REMP2 at 40 μmol/L). Results There were no significant difference in endotoxin-neutralizing percentages between PMB and REMP2 in concentrations of 0.10,10.00,100.00μmol/L(P>0.05).The contents of TNF-α were 1175±162,859±122,645±142,489±102 ng/L,respectively,after treatment of 10,20,40,80 μmol/L REMP2,which were obviously lower than that of NS(3463±218 ng/L,P<0.01).Under transmission electron microscope,the outer and inteior membranes of Eseherichia coli were obscure and rough,bacterial bodies were swollen with vacuoles in cytoplasm after treatment with REMP2.Conclusion REMP2 has ability of neutralizing endotoxin and also antibacterial activity.
