大黄素对血管紧张素Ⅱ诱导血管平滑肌细胞增殖的影响  

Inhibitory effects of emodin on proliferation of rat vascular smooth muscle cell induced by angiotensin Ⅱ

作　　者：杨雨民[1] 王世君[1] 金丹丹[2] 刘云英[2] 王兴祥[1] 陈君柱[1] 

机构地区：[1]浙江大学医学院附属第一医院心内科,浙江杭州310003 [2]浙江大学病原微生物实验室,浙江杭州310031

出　　处：《中国中药杂志》2008年第1期63-67,共5页China Journal of Chinese Materia Medica

基　　金：浙江省中医药重点项目研究计划(2007ZA015)

摘　　要：目的：观察大黄素对血管紧张素Ⅱ（AngⅡ）诱导血管平滑肌细胞（VSMC）增殖的影响及机制。方法：斑贴法培养大鼠主动脉VSMC,AngⅡ刺激VSMC建立细胞增殖模型。采用MTT法检测细胞增殖,观察大黄素（10,20,40,80μmol.L-1）、亚硝基-精氨酸甲酯（L-NAME,100μmol.L-1）对AngⅡ诱导VSMC增殖的影响。免疫组化法检测增殖细胞核抗原（PCNA）表达;硝酸还原酶及化学比色法测细胞上清液中一氧化氮（NO）、一氧化氮合酶（NOS）、诱导型一氧化氮合酶（iNOS）水平。半定量逆转录聚合酶链反应（RT-PCR）检测VSMC iNOS mRNA的表达。结果：大黄素在10~80μmol.L-1呈剂量及时间依赖性抑制VSMC增殖,但可被100μmol.L-1的L-NAME部分抵消;大黄素能减少PCNA阳性细胞表达,升高NO,NOS,iNOS水平,增加iNOS mRNA的表达。结论：大黄素对AngⅡ诱导的VSMC增殖有抑制作用,抑制VSMC PCNA的表达,上调iNOS基因表达,升高NO水平可能是其发挥作用的机制。Objective： To investigate the effects of emodin on the proliferation of cultured rat vascular smooth muscle cell（VSMC） induced by angiotensin II.Method： VSMCs were cultured by explant method.Cell proliferation model was established by stimulation with AngⅡ. Cell proliferation was measured by MTT assay to observe the effects of emodin（10,20,40 and 80 μmol·L-1） and NG-nitro-L-arginine methyl ester（L-NAME,100 μmol·L-1） on VSMC proliferation induced by AngⅡ.The expression of PCNA was measured by immunohistochemical staining.Nitric oxide（NO） level was measured by Griess reagent.Nitric oxide synthase（NOS） and inducible nitric oxide synthase（iNOS） levels were detected by chemical colorimetric method.mRNA expression of iNOS was measured by reverse transcription polymerase chain reaction（RT-PCR）.Result： Emodin at the doses range from 10 to 80 mol·L-1 inhibited cell proliferation in a dose and time-dependent manner.The inhibitory effects were partly blocked by 100 mol·L-1 of L-NAME.Emodin markedly decreased the expression of PCNA in VSMC,increased NO,NOS and iNOS levels,and increased iNOS mRNA expression in VSMC.Conclusion： Emodin could inhibite VSMCs proliferation induced by AngⅡ.Inhibiting the expression of PCNA,increasing the NO secretion and upregulating the iNOS gene expression might be associated with the inhibitory effects.

关 键 词：大黄素 血管紧张素Ⅱ 血管平滑肌细胞 一氧化氮 一氧化氮合酶 

分 类 号：R285.5[医药卫生—中药学]