miRNA-155靶向调控MMP-9对肝纤维化的作用及机制 .pdfVIP

miRNA-155靶向调控MMP-9对肝纤维化的作用及机制

李耕

【摘要】目的探讨微小RNA-155(miRNA-155)和基质金属蛋白酶-9(MMP-9)在

肝纤维化发生发展中的作用,为临床诊断和治疗肝纤维化提供依据.方法通过采用

miRNA-155抑制剂(miRNA-155inhibitor)、miRNA-155类似物(miRNA-155

mimic)及Wnt/β-catenin通路抑制剂处理肝星状细胞-T6(HSC-T6)细胞株,用荧

光定量聚合酶链式反应(PCR)检测miRNA-155、β-catenin及MMP-9的表达;采

用荧光素酶标记法检测MMP-9与miRNA-155相互作用的靶位点,探讨miRNA-

155表达变化与MMP-9和肝纤维化的关系.结果MMP-9在肝纤维化大鼠的肝脏

内表达显著上升,为正常大鼠的(8.51±0.30)倍;miRNA-155是与MMP-9相关的靶

向miRNA分子,其表达升高能显著降低MMP-9的表达;在使用Wnt通路抑制剂

后,miRNA-155和MMP-9的表达量显著降低.通过荧光素报告基因标记法发

现,miRNA-155与MMP-9的3-UTR靶向结合.结论miRNA-155是一种与肝纤维

化密切相关的miRNA分子,它可以通过影响MMP-9的表达来调节肝纤维化的发

生与发展.%ObjectiveToexploretheeffectofmiRNA-155andMMP9inthe

occurrenceanddevelopmentofliverfibrosistoprovidethebasisfor

clinicaldiagnosisandtreatmentofliverfibrosis.MethodsThemiRNA-155

inhibitor(miRNA-155inhibitor),miRNA-155analogues(miRNA-155mimic)

andWnt/β-cateninpathwayinhibitorwereadoptedtoprocessHSC-T6cell

lines,thefluorescencequantitativePCRwasusedtodetecttheexpressions

ofmiRNA-155,β-cateninandMMP-9.Andtheluciferaselabellingdetection

wasadoptedtodetectthetargetedpointofMMP-9andmiRNA-155

interactionforexploringtherelationshipbetweenmiRNA-155expression

changewithMMP-9andliverfibrosis.ResultsTheMMP-9expressioninthe

liverofhepaticfibrosisratswassignificantlyincreased,whichwas

(8.51±0.30)timesofnormalrats;miRNA-155wasatMMP-9-related

argetedmiRNAmoleculesitsexpressionincreasecouldsignificantly

decreasetheexpressionofMMP-9;afterusingtheWntpathway

inhibitor,theexpressionamountsofmiRNA-155andMMP-9were

significantlydecreased.Thefluoresceinreportgeneticmarkermethod

foundthatmiRNA-155wastargetedlycombinedwith3-UTRofMMP-

9