中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年
11期
1717-1721
,共5页
欧阳海春%吴沃栋%钟冬梅%吴焱贤%李文杰%陈晞明
歐暘海春%吳沃棟%鐘鼕梅%吳焱賢%李文傑%陳晞明
구양해춘%오옥동%종동매%오염현%리문걸%진희명
组织构建%血管内皮细胞%醒脑静%安宫牛黄丸%肿瘤坏死因子%内皮细胞%组织型纤溶酶原激活物%纤溶酶原激活物抑制剂1
組織構建%血管內皮細胞%醒腦靜%安宮牛黃汍%腫瘤壞死因子%內皮細胞%組織型纖溶酶原激活物%纖溶酶原激活物抑製劑1
조직구건%혈관내피세포%성뇌정%안궁우황환%종류배사인자%내피세포%조직형섬용매원격활물%섬용매원격활물억제제1
Drugs,Chinese Herbal%Tumor Necrosis Factor-alpha%Tissue Plasminogen Activator
背景:前期研究发现醒脑静能有效抑制兔心肌缺血再灌注模型的炎症递质及促纤溶作用,但是影响纤溶系统活性的作用机制未完全明确。目的:观察醒脑静对重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1及其基因表达的影响。方法:取3-5代人脐静脉内皮细胞,在培养基中添加10μg/L 重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1及其基因表达,醒脑静组加入不同浓度(5,10,20 mL/L)的醒脑静干预,阳性对照组添加氟伐他汀(1μmol/L),并设立单纯人脐静脉内皮细胞培养的空白对照组。培育24 h后采用酶联免疫吸附双抗体夹心法(ELISA)检测细胞上清液组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1水平;采用反转录聚合酶链反应检测人脐静脉内皮细胞的组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1的mRNA表达。结果与结论:重组人肿瘤坏死因子α组纤溶酶原激活物抑制剂1分泌和 mRNA 表达较空白对照组显著升高(P <0.05),组织型纤溶酶原激活物分泌和mRNA表达较空白对照组显著降低(P <0.05)。不同浓度醒脑静组纤溶酶原激活物抑制剂1分泌和mRNA表达均较重组人肿瘤坏死因子α组显著降低(P <0.05),而组织型纤溶酶原激活物分泌和mRNA表达均较重组人肿瘤坏死因子α组显著升高(P <0.05),且呈剂量依赖关系。结果证实,醒脑静作用可逆转重组人肿瘤坏死因子α所致的人脐静脉内皮细胞的纤溶活性。
揹景:前期研究髮現醒腦靜能有效抑製兔心肌缺血再灌註模型的炎癥遞質及促纖溶作用,但是影響纖溶繫統活性的作用機製未完全明確。目的:觀察醒腦靜對重組人腫瘤壞死因子α介導人臍靜脈內皮細胞分泌組織型纖溶酶原激活物和纖溶酶原激活物抑製劑1及其基因錶達的影響。方法:取3-5代人臍靜脈內皮細胞,在培養基中添加10μg/L 重組人腫瘤壞死因子α介導人臍靜脈內皮細胞分泌組織型纖溶酶原激活物和纖溶酶原激活物抑製劑1及其基因錶達,醒腦靜組加入不同濃度(5,10,20 mL/L)的醒腦靜榦預,暘性對照組添加氟伐他汀(1μmol/L),併設立單純人臍靜脈內皮細胞培養的空白對照組。培育24 h後採用酶聯免疫吸附雙抗體夾心法(ELISA)檢測細胞上清液組織型纖溶酶原激活物和纖溶酶原激活物抑製劑1水平;採用反轉錄聚閤酶鏈反應檢測人臍靜脈內皮細胞的組織型纖溶酶原激活物和纖溶酶原激活物抑製劑1的mRNA錶達。結果與結論:重組人腫瘤壞死因子α組纖溶酶原激活物抑製劑1分泌和 mRNA 錶達較空白對照組顯著升高(P <0.05),組織型纖溶酶原激活物分泌和mRNA錶達較空白對照組顯著降低(P <0.05)。不同濃度醒腦靜組纖溶酶原激活物抑製劑1分泌和mRNA錶達均較重組人腫瘤壞死因子α組顯著降低(P <0.05),而組織型纖溶酶原激活物分泌和mRNA錶達均較重組人腫瘤壞死因子α組顯著升高(P <0.05),且呈劑量依賴關繫。結果證實,醒腦靜作用可逆轉重組人腫瘤壞死因子α所緻的人臍靜脈內皮細胞的纖溶活性。
배경:전기연구발현성뇌정능유효억제토심기결혈재관주모형적염증체질급촉섬용작용,단시영향섬용계통활성적작용궤제미완전명학。목적:관찰성뇌정대중조인종류배사인자α개도인제정맥내피세포분비조직형섬용매원격활물화섬용매원격활물억제제1급기기인표체적영향。방법:취3-5대인제정맥내피세포,재배양기중첨가10μg/L 중조인종류배사인자α개도인제정맥내피세포분비조직형섬용매원격활물화섬용매원격활물억제제1급기기인표체,성뇌정조가입불동농도(5,10,20 mL/L)적성뇌정간예,양성대조조첨가불벌타정(1μmol/L),병설립단순인제정맥내피세포배양적공백대조조。배육24 h후채용매련면역흡부쌍항체협심법(ELISA)검측세포상청액조직형섬용매원격활물화섬용매원격활물억제제1수평;채용반전록취합매련반응검측인제정맥내피세포적조직형섬용매원격활물화섬용매원격활물억제제1적mRNA표체。결과여결론:중조인종류배사인자α조섬용매원격활물억제제1분비화 mRNA 표체교공백대조조현저승고(P <0.05),조직형섬용매원격활물분비화mRNA표체교공백대조조현저강저(P <0.05)。불동농도성뇌정조섬용매원격활물억제제1분비화mRNA표체균교중조인종류배사인자α조현저강저(P <0.05),이조직형섬용매원격활물분비화mRNA표체균교중조인종류배사인자α조현저승고(P <0.05),차정제량의뢰관계。결과증실,성뇌정작용가역전중조인종류배사인자α소치적인제정맥내피세포적섬용활성。
BACKGROUND:We have found thatXingnaojing can effectively inhibit mediators of inflammatory mediators and promote fibrinolytic activity in a rabbit model of myocardial ischemia and reperfusion. However, it is not completely clear what is the action of mechanism of fibrinolytic system activity in human umbilical vein endothelial cels.
<br> OBJECTIVE:To investigate the effects ofXingnaojingon recombinant human tumor necrosis factor α-mediated tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) expression in human umbilical cord vein endothelial cels. METHODS: The passages 3-5 of human umbilical cord vein endothelial cels were obtained, and were induced by adding 10 μg/L recombinant human tumor necrosis factor α in the culture medium.Xingnaojing in different concentrations (5, 10, 20 mL/L) were injected in theXingnaojing group, and fluvastatin (1 μmol/L) was added as positive control group. A blank control group of simple human umbilical cord vein endothelial cels culture was set up. After 24 hours of cultivation, the production of t-PA and PAI-1 in the cultured medium was determined by ELISA. The mRNA expression of t-PA and PAI-1 was measured by reverse transcript-PCR. RESULTS AND CONCLUSION:Compared with the blank control group, the secretion and mRNA expression of PAI-1 in the group treated with recombinant human tumor necrosis factor α(10μg/L) were significantly increased (P < 0.05); however, the secretion and mRNA expression of t-PA in the group treated with recombinant human tumor necrosis factorα (10μg/L) were significantly decreased (P < 0.05). The secretion and mRNA expression of t-PA were significantly higher in theXingnaojinggroup at different concentrations than in the recombinant human tumor necrosis factor α group (P < 0.05); however, the secretion and mRNA expression of PAI-1 were significantly lower in the former group than the latter group (P< 0.05), which was in a dose-dependent manner.Xingnaojing can effectively improve the fibrinolytic function of human umbilical cord vein endothelial cels mediated by recombinant human tumor necrosis factor α.