miR-181a對牛骨骼肌衛(wèi)星細(xì)胞分化的影響
發(fā)布時間:2018-04-05 01:00
本文選題:microRNA-a(miR-a) 切入點:牛 出處:《黑龍江畜牧獸醫(yī)》2017年21期
【摘要】:為了探討miR-181a對牛骨骼肌衛(wèi)星細(xì)胞分化的影響,試驗將分離培養(yǎng)的牛骨骼肌衛(wèi)星細(xì)胞接種在6孔板中,待細(xì)胞匯合度為70%時,用PEI轉(zhuǎn)染miR-181a的模擬物miR-181a-M、抑制劑miR-181a-I、模擬物對照miR-181a-M-NC和抑制劑對照miR-181a-I-NC進(jìn)入牛骨骼肌衛(wèi)星細(xì)胞中,在0,1,2,3天用Western-blot檢測肌肉有關(guān)分化蛋白MyoD、MyoG和MYH3以及靶基因HOX-A11蛋白的表達(dá)量,用免疫熒光法檢測細(xì)胞肌管的數(shù)量。結(jié)果表明:在牛骨骼肌衛(wèi)星細(xì)胞中,轉(zhuǎn)染miR-181a模擬物的牛骨骼肌衛(wèi)星細(xì)胞,有關(guān)肌肉分化蛋白質(zhì)如MyoD、MyoG和MYH3的表達(dá)量明顯增加,細(xì)胞肌管數(shù)量顯著增多,進(jìn)一步證實miR-181a促進(jìn)細(xì)胞的分化。在雙熒光素酶報告系統(tǒng)中,miRNA-181a和HOX-A11基因的3'-UTR結(jié)合時熒光素酶活性明顯下降,miRNA-181a和突變的3'-UTR結(jié)合時熒光素酶活性沒有下降。說明HOX-A11是miR-181a的靶基因,miR-181a對牛骨骼肌衛(wèi)星細(xì)胞的分化具有促進(jìn)作用。
[Abstract]:In order to investigate the effect of miR-181a on the differentiation of bovine skeletal muscle satellite cells, the isolated and cultured bovine skeletal muscle satellite cells were inoculated in a 6-well plate.鐢≒EI杞煋miR-181a鐨勬ā鎷熺墿miR-181a-M,鎶戝埗鍓俶iR-181a-I,妯℃嫙鐗╁鐓iR-181a-M-NC鍜屾姂鍒跺墏瀵圭収miR-181a-I-NC榪涘叆鐗涢楠艱倢鍗槦緇嗚優(yōu)涓,
本文編號:1712466
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