摘要
目的:探讨microRNA-21( miR-21)反义寡核苷酸对肝星状细胞胶原合成的影响。方法肝星状细胞分离和培养,利用脂质体2000将miR-21反义寡核苷酸转染活化的肝星状细胞中,转染48 h后,收集肝星状细胞,采用实时RT-PCR法和Western blotting分别检测细胞miR-21表达和Ⅰ、Ⅲ型胶原蛋白表达;MTT法检测转染后细胞增殖活性。结果与无关反义寡核苷酸组相比,miR-21反义寡核苷酸转染后miR-21表达较无关对照组下调约76%( P <0.01);细胞增殖活性降低(26±3)%( P <0.01);Ⅰ、Ⅲ型胶原蛋白分别降低(61±7)%和(48±6)%(均P <0.01)。结论 miR-21反义寡核苷酸能显著下调miR-21的表达,抑制肝星状细胞增殖及胶原的合成。
Objective To investigate the effect of microRNA-21 (miR-21) antisense oligonucleoti-de on collagen synthesis in the rat hepatic satellite cells ( HSCs) .Methods Rat hepatic stellate cells were isolated and cultured; the miR-21 antisense oligonucleotide was transfected into HSCs with lipofectamine 2000;after incubation 48 h, the HSCs were collected.The expression of miR-21 was detected with reverse transcription polymerase chain reaction ( RT-PCR) , andα-smooth muscle actin (α-SMA) protein in HSCs with Western blot.The cell proliferation was assayed with methyl thiazolyl tetrazolium ( MTT) method.Re-sults Compared to scrambled control group, the expression of miR-21 was reduced by 76%( P 〈0.01), the proliferation activity of HSCs was reduced by(26 ±3)%( P 〈0.01), the expressions of type I and III collagen proteins were reduced by(61 ±7)%and (48 ±6)%( P 〈0.01).Conclusions The miR-21 an-tisense oligonucleotide could reduce significantly the expression of miR-21, and inhibit HSC proliferation and extracellular matrix production.
出处
《中国医师杂志》
CAS
2015年第3期337-339,共3页
Journal of Chinese Physician
基金
浙江省自然科学基金资助项目(LY14H030010)
温州市科技局资助项目(Y20120032)