本文選題：二葉式主動(dòng)脈瓣 + 瓣膜鈣化��； 參考：《南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版)》2017年09期

【摘要】：目的 :探究二葉式主動(dòng)脈瓣中miRNA-195調(diào)控瓣膜鈣化的可能機(jī)制。方法 :收集35例接受主動(dòng)脈瓣置換術(shù)患者狹窄的二葉式或三葉式主動(dòng)脈瓣,通過(guò)PCR及Western blot分別檢測(cè)miRNA-195的表達(dá)量、果蠅母源抗皮膚生長(zhǎng)因子(drosophila mothers against decapentaplegic 7,SMAD7)的mRNA及蛋白表達(dá)量。雙熒光素酶法預(yù)測(cè)miRNA-195的靶基因。在豬瓣膜間質(zhì)細(xì)胞中分別沉默和過(guò)表達(dá)miRNA-195,檢測(cè)SMAD7的mRNA表達(dá)水平并探究?jī)烧叩南嗷リP(guān)系。在人類(lèi)瓣膜組織上通過(guò)免疫組化染色檢測(cè)基質(zhì)金屬蛋白酶(matrix metalloproteinase,MMP)-2、MMP-9蛋白水平,通過(guò)天狼星紅染色檢測(cè)膠原蛋白水平,通過(guò)Von Kossa染色比較鈣化程度差異,進(jìn)而研究鈣化相關(guān)的功能改變。結(jié)果 :與狹窄的三葉式主動(dòng)脈瓣相比,狹窄的二葉式主動(dòng)脈瓣中miRNA-195表達(dá)降低,SMAD7的mRNA和蛋白水平升高。雙熒光素酶實(shí)驗(yàn)提示SMAD7是miRNA-195的直接靶標(biāo)。在豬瓣膜間質(zhì)細(xì)胞中沉默miRNA-195引起SMAD7 mRNA水平升高,過(guò)表達(dá)miRNA-195引起SMAD7 mRNA水平降低。狹窄鈣化的二葉式主動(dòng)脈瓣組織中MMP-2、MMP-9及膠原的表達(dá)較狹窄鈣化的三葉式主動(dòng)脈瓣升高。結(jié)論:在二葉式主動(dòng)脈瓣中,下調(diào)的miRNA-195促進(jìn)了SMAD7表達(dá),進(jìn)而促進(jìn)細(xì)胞外基質(zhì)的纖維化并最終促進(jìn)其鈣化。
[Abstract]:Aim: to explore the possible mechanism of miRNA-195 regulating valve calcification in two-leaf aortic valve. Methods: the expression of miRNA-195 and the expression of (drosophila mothers against decapentaplegic 7 SMAD7 were detected by PCR and Western blot in 35 patients with stenosis of aortic valve. The target gene of miRNA-195 was predicted by double luciferase method. MiRNA-195 was silenced and overexpressed in porcine valvular interstitial cells to detect the mRNA expression of SMAD7 and to explore the relationship between them. The levels of matrix metalloproteinase (MMP) -2MMP-9, collagen and calcification were detected by immunohistochemical staining, Sirius red staining and Von Kossa staining respectively. Furthermore, the functional changes associated with calcification were studied. Results: the expression of miRNA-195 in the stenosis two-leaf aortic valve decreased the mRNA and protein levels of SMAD7 compared with the stenosis trefoil aortic valve. Double luciferase assay suggested that SMAD 7 was a direct target for miRNA-195. Silencing miRNA-195 increased the level of SMAD7 mRNA in porcine valve interstitial cells and decreased the level of SMAD7 mRNA induced by overexpression of miRNA-195. The expression of MMP-2, MMP-9 and collagen in two-lobe aortic valve with stenosis calcification was higher than that in trefoil aortic valve with narrow calcification. Conclusion: in two-leaf aortic valve, down-regulated miRNA-195 promotes the expression of SMAD7, thus promotes the fibrosis of extracellular matrix and ultimately promotes its calcification.
【作者單位】： 南京醫(yī)科大學(xué)第一附屬醫(yī)院心臟大血管外科;
【基金】：江蘇省自然科學(xué)基金(BK20151590)
【分類(lèi)號(hào)】：R654.2

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