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作 者:贾本川 郭冰玉 李泰然 邹日峰 刘春雨 于兵 陶凯 JIA Ben-chuan;GUO Bing-yu;LI Tai-ran;ZOU Ri-feng;LIU Chun-yu;YU Bing;TAO Kai(Reconstructive and Plastic Surgery,General Hospital of Northern Theater Command,Shenyang 110016,China)
机构地区:[1]北部战区总医院烧伤整形科,辽宁沈阳110016
出 处:《解剖科学进展》2021年第2期218-221,225,共5页Progress of Anatomical Sciences
基 金:辽宁省自然科学基金(20180140211)。
摘 要:目的探讨miR-663调节TGF-β1表达抑制增生性瘢痕皮肤成纤维细胞增殖的作用及机制。方法RT-PCR检测组织中miR-663和TGF-β1mRNA的表达;MIRDB软件预测、双荧光素酶报告基因实验分析miR-663对TGF-β1基因的靶向作用;MTT检测miR-663的差异性表达对成纤维细胞增殖的影响;Western blot检测miR-663差异性表达对成纤维细胞TGF-β1和Smad3表达的影响。结果增生性瘢痕组织中miR-663的表达明显低于正常瘢痕组织和正常皮肤组织,TGF-β1的mRNA在增生性瘢痕组织中表达上调;荧光素酶报告基因实验显示miR-663对TGF-β1基因的靶向作用;miR-663过表达能显著抑制成纤维细胞的增殖,miR-663 inhibitor能够明显促进成纤维细胞的增殖;miR-663过表达能够抑制成纤维细胞TGF-β1和Smad3的表达,而miR-663 inhibitor则使其TGF-β1和Smad3的表达上调。结论 miR-663通过抑制TGF-β1进而抑制成纤维细胞的增殖。Objective To investigate the effect and mechanism of miR-663 regulating the expression of TGF-β 1 and inhibiting the proliferation of hypertrophic scar skin fibroblasts. Methods RT-PCR was used to detect the expression of miR-663 and mRNA of TGF-β 1. MIRDB software was used to predict the binding sites of miR-663 and TGF-β1, and the target effect of miR-663 on TGF-β1 was detected by luciferase reporter gene experiment. MTT test was used to detect the effect of miR-663 on cell proliferation. Western blot was used to detect the effect of miR-663 on the expression of TGF-β1 and Smad3. Results The expression of miR-663 in hypertrophic scar tissue was lower than that in normal scar tissue and normal skin tissue, however the expression of TGF-β1 m RNA was higher in hypertrophic scar tissues. MIRDB showed that there were binding sites in the 3’ noncoding region of miR-663 and TGF-β1. Luciferase reporter gene experiment proved that miR-663 could directly reduce the activity of TGF-β1. Overexpression of miR-663 can significantly inhibit the proliferation of fibroblasts, miR-663 inhibitor can significantly promote the proliferation of fibroblasts;miR-663 overexpression can inhibit the expression of TGF-β 1 and Smad3, while miR-663 inhibitor can up regulate the expression of TGF-β 1 and Smad3. Conclusion miR-663 can inhibit the growth of fibroblasts by inhibiting TGF-β1.
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