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时间:2020-05-20
《辛伐他汀对肿瘤坏死因子α诱导的人腹膜间皮细胞组织型纤溶酶原激活物和纤溶酶原激活物抑制因子1表达的影响.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、临床肾脏病杂志2015年4月第l5卷第4期JClinNephrol,April2015,Vo1.15,No.4·239··实验研究·辛伐他汀对肿瘤坏死因子诱导的人腹膜间皮细胞组织型纤溶酶原激活物和纤溶酶原激活物抑制因子1表达的影响田功何劲松【摘要】目的探讨辛伐他汀对体外培养的人腹膜间皮细胞(humanperitonealmesothelialcells,HPMCs)在肿瘤坏死因子a(tumornecrosisfactor-a,TNF_a)诱导后组织型纤溶酶原激活物(tis-sue-typeplasminogenactivator,t-PA)和纤溶酶原激活
2、物抑制因子1(plasminogenactivatorinhibitor-1,PAId)表达的影响。方法应用胰蛋白酶消化法分离HPMCs进行原代培养并传代,将第三代分为5组(每组设3个样本):①正常对照组:将HPMCs置于完全培养液中,置于37℃、5CCh培养箱培养24h;②单纯TNF-a(I~g/L)诱导组:将HPMCs置于浓度为l~g/L的TNF-~诱导缓冲液中,环境温度及处理时间如前;③TNF-a诱导+辛伐他汀(2.5pmol/L)组:将HPMCs置于含有TNF-a(1gg/L)诱导液+辛伐他汀(2.5/lmol/L)缓冲液中,余同前;④TNF-~
3、诱导+辛伐他汀(5.0gmol/L)组:将HPMCs置于含有TNF-a(1t~g/L)诱导液+辛伐他汀(5.0/~mol/L)缓冲液中,余同前;⑤TNF-u诱导+辛伐他汀(10.0~mol/L)组:将HPMCs置于含有TNF-a(1~g/L)诱导液+辛伐他汀(10.0pmol/L)缓冲液中,余同前。采用半定量逆转录聚合酶链反应(reversetranscription—polymerasechainreaction,RT-PCR)法检测细胞内t_PA和PAI-1mRNA表达;酶联免疫吸附法(enzyme-linkedimmunosorbentassay,
4、ELISA)检测细胞上清液中t-PA和PAI_1的蛋白质水平,用二喹啉甲酸(bicin—choninicacid,BCA)蛋白检测法测定细胞中蛋白质含量,用以校正ELISA结果。结果与正常对照组比较,TNF-a诱导能抑制HPMCs中t-PA表达,增加PAI_1的表达(P<0.01);与单纯TNF-a诱导组比较,辛伐他汀(2.5~mol/L)能显著增加TNF_a诱导HPMCs中t-PA表达,抑制PAI-1的表达(P5、mol/L)亦能明显增加TNF-a诱导的HPMCs中t-PA表达,抑制PAI_1的表达(P6、sothelialcellsinvitroTIANGong,HEJin-song.DepartrrwmtofNephrology,theAffiliatedDrumTowerHospital,NanjingUniversityIVIedicalSchool,njing210008,ChinaCorrespondingauthor:HeJin-song,F-mail:^insonmedmail.corn.c[Abstract]ObjectiveToinvestigatetheeffectsofSimvastatinontissue-typeplasmino7、genacti—vator(PA)andplasminogenactivatorinhibitor-1(PAI-1)expressioninhumanperitonea1mesotheli—a1cells(HPMCs)culturedunderinflammatoryconditions.MethodsHPMCswereisolatedfromhu—manomentabytrypsindigestionmethodandsubcultured.Then,thethird-generationHPMCsweredividedinto5groups:norm8、alcontrolgroup(TheHPM.Cswereculturedinth
5、mol/L)亦能明显增加TNF-a诱导的HPMCs中t-PA表达,抑制PAI_1的表达(P6、sothelialcellsinvitroTIANGong,HEJin-song.DepartrrwmtofNephrology,theAffiliatedDrumTowerHospital,NanjingUniversityIVIedicalSchool,njing210008,ChinaCorrespondingauthor:HeJin-song,F-mail:^insonmedmail.corn.c[Abstract]ObjectiveToinvestigatetheeffectsofSimvastatinontissue-typeplasmino7、genacti—vator(PA)andplasminogenactivatorinhibitor-1(PAI-1)expressioninhumanperitonea1mesotheli—a1cells(HPMCs)culturedunderinflammatoryconditions.MethodsHPMCswereisolatedfromhu—manomentabytrypsindigestionmethodandsubcultured.Then,thethird-generationHPMCsweredividedinto5groups:norm8、alcontrolgroup(TheHPM.Cswereculturedinth
6、sothelialcellsinvitroTIANGong,HEJin-song.DepartrrwmtofNephrology,theAffiliatedDrumTowerHospital,NanjingUniversityIVIedicalSchool,njing210008,ChinaCorrespondingauthor:HeJin-song,F-mail:^insonmedmail.corn.c[Abstract]ObjectiveToinvestigatetheeffectsofSimvastatinontissue-typeplasmino
7、genacti—vator(PA)andplasminogenactivatorinhibitor-1(PAI-1)expressioninhumanperitonea1mesotheli—a1cells(HPMCs)culturedunderinflammatoryconditions.MethodsHPMCswereisolatedfromhu—manomentabytrypsindigestionmethodandsubcultured.Then,thethird-generationHPMCsweredividedinto5groups:norm
8、alcontrolgroup(TheHPM.Cswereculturedinth
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