[1]孙亚丽,郭 华,马建洲.内毒素结合肽P1和P2抗内毒素活性的实验研究[J].现代检验医学杂志,2017,32(06):81-84.[doi:10.3969/j.issn.1671-7414.2017.06.001]
　SUN Ya-li,GUO Hua,MA Jian-zhou.Study on the Anti-Endotoxin Activity of Human Endotoxin Binding Peptide P1 and P2[J].Journal of Modern Laboratory Medicine,2017,32(06):81-84.[doi:10.3969/j.issn.1671-7414.2017.06.001]
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内毒素结合肽P1和P2抗内毒素活性的实验研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第32卷
期数:
2017年06期
页码:
81-84
栏目:
论著
出版日期:
2017-12-25

文章信息/Info

Title:
Study on the Anti-Endotoxin Activity of Human Endotoxin Binding Peptide P1 and P2
文章编号:
1671-7414(2017)06-081-04
作者:
孙亚丽1; 郭 华1; 马建洲2
1.陕西省人民医院病理科,西安 710068; 2.西安解放军第323医院眼科,西安 710054
Author(s):
SUN Ya-li1; GUO Hua1; MA Jian-zhou2
1.Department of Pathology,Shaanxi Provincial People's Hospital,Xi'an 710068,China; 2.Department of Ophthalmology,the PLA 323th Hospital,Xi'an 710054,China
关键词:
脂多糖;  内毒素结合肽P1;  内毒素结合肽P2;  抗内毒素活性
分类号:
R-332
DOI:
10.3969/j.issn.1671-7414.2017.06.001
文献标志码:
A
摘要:
目的 研究基于脂多糖(lipolysachride,LPS)结合蛋白的新型内毒素结合肽P1和P2的抗内毒素活性。方法 设计并合成内毒素结合肽P1和P2。体外实验取一名健康志愿者静脉血100 ml分离的外周血单个核细胞(PBM),分为正常对照组、LPS组、阳性对照组LPS(1 mg/L)+多黏菌素B(12.5 mg/L)、P1组LPS(1 mg/L)+P1(2 mg/L,5 mg/L,12.5 mg/L)、P2组LPS(1 mg/L)+P2(2 mg/L,5 mg/L,12.5 mg/L)。ELISA方法检测各组上清液中TNF-α和IL-6的浓度。体内实验将40只昆明小鼠平均分为四组,每组10只,分别为空白对照组、LPS模型组、LPS+P1治疗组和LPS+P2治疗组。取各组小鼠肺脏和肝脏组织制作HE切片,观察组织形态学变化。结果 ①与1 mg/L LPS比较,1 mg/L LPS+12.5 mg/L的P1以及1 mg/L LPS+3种浓度P2刺激后,PBMC培养上清液中IL-6及TNF-α的水平均明显降低,差异均有统计学意义(均P<0.01),12.5 mg/L的P2与阳性对照PMBC作用后上清液中TNF-α浓度比较,差异无统计学意义(P>0.05)。②组织形态学显示,模型组小鼠肝小叶中央静脉及肝窦扩张充血,肝细胞浊肿,偶见肝细胞嗜酸性变及点状坏死。肺间质充血水肿,灶性出血,肺泡腔狭小。10 mg/kg P1处理组肝细胞浊肿较模型组减轻,肺间质充血水肿减轻。10 mg/kg P2处理组,肝小叶中央静脉及肝窦充血程度减轻,肺间质充血水肿明显减轻。结论 体内外实验表明合成内毒素结合肽P1和P2对LPS导致的机体损伤具有拮抗作用。体外实验结果还表明,12.5 mg/L P2对TNF-α或IL-6的释放抑制作用较为肯定。
Abstract:
Abstract:Objective To study the antiendotoxin activity of P1 and P2 based on the lipopolysacchride binding protein.MethodP1 and P2 were designed and obtained.In vitro test,peripheral bloodmononuclear cell(PBMC)were extracted from volunteer 100ml venous blood,the experiment group was arranged as:control group,positive control group,LPS group,LPS+P1(2 mg/L,5 mg/L,12.5 mg/L)group and LPS+P2(2 mg/L,5 mg/L,12.5 mg/L)group,TNF-α and IL-6 of supernatant liquor in every group were detectdby ELISA.In vivo,40 kunming rice were randomly divided into four groups with ten rice each group:control group,model group,LPS+P1 and LPS+P2 group,Pathologic changes of lung and liver tissues were observed by hematoxylin and eosin(HE)staining.Results The serum level of IL-6 and TNF-αin 12.5 mg/L P1 and 2 mg/L,5 mg/L,12.5 mg/L P2 treatment group were lower than that in model group,the difference was statistically significant(all P<0.01).Serum level of TNF-α or IL-6 in 12.5 mg/L P2 treatment group were similar to that in PMB treatment group,and there was no statistically significantdifference(P>0.05).Histologymorphology finding showed that central veinsof liver and hepatic sinusoid congestion,hepatic cellular edema existed,occasionally,acidophilic change and spotty necrosis were found,pulmonary interstitial edema,focal hemorrhage,alveolar space stenosis existed.As regards 10 mg/kgP1 treatment group mice,hepatic cellular edema and pulmonary interstitial edema ameliorated.About 10 mg/kg P2 treatment group mice,veins of liver and hepatic sinusoid congestion obviously ameliorated,mild pulmonary interstitial edemaexsited.Conclusion The results indicated P1 and P2 had antiendotoxin effect,in vivo and vitro,for 12.5 mg/L P2,its inhibition effectfor TNF-α or IL-6 release was positive.

参考文献/References:

[1] Ryu JK,Kim SJ,Rah SH,et al.Reconstruction of LPS transfer cascade reveals structural determinants within LBP,CD14,and TLR4-MD2 for efficient LPS recognition and transfer[J].Immunity,2017,46(1):38-50.
[2] Kim SJ,Kim HM.Dynamic lipopolysaccharide transfer cascade to TLR4/MD2 complex via LBP and CD14[J].BMB Rep,2017,50(2):55-57.
[3] Lamping N,Hoess A,Yu B,et al.Effect of site-directed mutagenesis ofbasic residues(Arg94,Lys95,Lys99)of lipopolysaccharide(LPS)-binding proteinon binding and transfer of LPS and subsequent immune cell activation[J].Jounal of Immunology,1996,157(10):4648-4656.
[4] Guo S,Li S,Li F,et,al.Modification of a synthetic LPS-binding domain of anti-lipopolysaccharide factor from shrimp reveals strong structure activity relationship in their antimicrobial characteristics[J].Dev Comp Immunol,2014,45(2):227-232.
[5] Nan YH,Bang JK,Jacob B,et al.Prokaryotic selectivity and LPS-neutralizing activity of short antimicrobial peptides designed from the human antimicrobial peptide LL-37[J].Peptides,2012,35(2):239-247.
[6] Santos DES,Pol-Fachin L,Lins RD,et al.Polymyxin binding to the bacterial outer membrane reveals cation displacement and increasing membrane curvature in susceptible but not in resistant lipopolysaccharide chemotypes[J].J ChemInf Model,2017,57(9):2181-2193. 收稿日期:2017-04-26 修回日期:2017-11-01

备注/Memo

备注/Memo:
基金项目:陕西省自然科学基金,资助项目编号2010JQ4026。 作者简介:孙亚丽(1975-),女,研究生,主治医师,研究方向:肿瘤病理,E-mail:ylisun@163.com。 通讯作者:马建洲(1973-),男,研究生,主治医师,研究方向为眼科肿瘤,E-mail:153968487@qq.com。
更新日期/Last Update: 2017-12-26