非抗凝肝素衍生物ROH对脓毒症小鼠脑损伤的保护作用及机制研究  

Protective effect and mechanism of non-anticoagulant heparin derivative ROH on brain injury in septic mice

作　　者：张嘉茜 赵晓禹 梁洋 崔慧斐[1] ZHANG Jia-xi;ZHAO Xiao-yu;LIANG Yang;CUI Hui-fei(School of Pharmacy, Shandong University, Jinan 250012, Shandong Province, China;Clinical Trial Research Center,Jinan Central Hospital Affiliated to Shandong University,Jinan 250013,Shandong Province,China)

机构地区：[1]山东大学药学院,山东济南250012 [2]山东大学附属济南市中心医院临床试验研究中心,山东济南250013

出　　处：《中国临床药理学杂志》2019年第13期1352-1355,共4页The Chinese Journal of Clinical Pharmacology

基　　金：山东省科技重大专项(重大关键技术)基金资助项目(2015ZDJS04002)

摘　　要：目的研究非抗凝肝素衍生物--ROH对脂多糖(LPS)诱导的脓毒症小鼠脑损伤的保护作用及机制。方法按数表法将Balb/c小鼠随机分为4组:正常组、模型组、对照组和实验组。小鼠腹腔注射LPS 10 mg·kg^-1建立脓毒症急性脑损伤模型。对照组和实验组在造模前15 min,分别尾静脉注射肝素2. 5mg·kg^-1、ROH 7. 5 mg·kg^-1进行干预,正常组注射等体积0. 9%Na Cl。以免疫组化法检测脑组织乙酰肝素酶(HPA)和金属基质蛋白酶-9(MMP-9)表达水平;伊文思蓝染色法检测血脑屏障通透性;TBA法测定脑组织中丙二醛(MDA)的含量。结果正常组、模型组、对照组和实验组的HPA表达水平(光密度值)分别为(8. 0±8. 0)×10^-5,(3. 0±1. 8)×10^-3,(2. 3±1. 1)×10^-4和(5. 4±3. 3)×10^-4;上述这4组的MMP-9表达水平(光密度值)分别为(8. 0±8. 0)×10^-5,(3. 6±1. 8)×10^-3,(3. 9±1. 4)×10^-4和(4. 9±3. 3)×10^-4;上述这4组的脑内伊文思蓝的含量分别为(4. 36±1. 58),(78. 60±14. 75),(36. 28±9. 16)和(44. 09±8. 95)μg·g^-1;上述这4组的MDA含量分别为(6. 91±1. 73),(10. 88±2. 08),(6. 68±1. 58)和(5. 41±1. 50)nmol·mL^-1。模型组与正常组相比,上述指标的差异均有统计学意义(P <0. 05,P <0. 01);对照组和实验组与模型组相比,上述指标的差异均有统计学意义(P <0. 05,P <0. 01)。结论 ROH与肝素一样,通过抑制脓毒症时LPS所致的HPA和MMP-9高表达,保护内皮糖萼的完整性,从而发挥脑保护作用,并提高脓毒症小鼠的生存率。Objective To study the protective effect and mechanism of non - anticoagulated heparin derivatives reduced oxyheparin ( ROH) on brain damage induced by lipopolysaccharide ( LPS ) in septic mice. Methods The Balb /c mice were randomly divided into 4 groups: normal group,model group,control group,and experimental group. LPS ( 10 mg · kg^-1 ) was injected by intraperitoneal injection to establish acute brain injury septic mice model . Heparin ( 2. 5 mg·kg^-1 ) and ROH( 7. 5 mg·kg^-1 ) were respectively injected by intravenous injention in control group and experimental group at the time of 15 min before modeling,and normal group was injected with the same volume of normal saline. Immunohistochemistry was used to detect the expression of heparanase ( HPA) and matrix metalloproteinase - 9 ( MMP - 9) in brain tissue. Evans blue staining was used to detect blood - brain barrier permeability. TBA method was used to measure the content of dialdehyde ( MDA) in brain tissue. Results The expression level( optical density,OD) of HPA in normal group,model group,control group and experimental group were ( 8. 0 ± 8. 0 )× 10^-5,( 3. 0 ± 1. 8 )× 10^-3,( 2. 3 ± 1. 1)× 10^-4,and ( 5. 4 ± 3. 3 )× 10^-4;the expression level ( OD) of MMP - 9 in these four groups were ( 8. 0 ± 8. 0)× 10^-5,( 3. 6 ± 1. 8)× 10^-3,( 3. 9 ± 1. 4)× 10^-4,and( 4. 9 ± 3. 3)× 10^-4;the contents of evans blue in these four groups were( 4. 36 ± 1. 58),( 78. 60 ± 14. 75),( 36. 28 ± 9. 16),and( 44. 09 ± 8. 95)μg·g^-1;the concentrations of MDA in these four groups were ( 6. 91 ± 1. 73 ),( 10. 88 ± 2. 08 ),( 6. 68 ± 1. 58 ),( 5. 41 ± 1. 50 ) nmol·mL^-1. Comparison between model group and normal group,the difference of the factors were significantly ( P < 0. 05,P < 0. 01);comparison between experimental group and control group with model group,the difference of the factors were significantly( P < 0. 05,P < 0. 01). Conclusion ROH,like heparin,protects the integrity of endothelial glycosides by inhibiting the high expression of HPA and MMP

关 键 词：脓毒症 血脑屏障 内皮糖萼 非抗凝肝素 

分 类 号：R972[医药卫生—药品]