miR-2116-3p在人增生性瘢痕中的表达及作用
R446%R318%R62; 背景:目前多项研究证实了miRNA影响增生性瘢痕的发生发展,Ⅰ型胶原与增生性瘢痕成纤维细胞密切相关,猜测miR-2116-3p也有可能与增生性瘢痕的发生发展有关系.目的:探讨miR-2116-3p在人增生性瘢痕中的表达及作用.方法:收集新疆医科大学第一附属医院6例患者的增生性瘢痕组织与6例患者重睑术后正常皮肤组织,采用qRT-PCR法检测miR-2116-3p与Ⅰ型胶原的表达.取增生性瘢痕组织,原代培养第3-6代成纤维细胞,分为阴性对照组、miR-2116-3p模拟物组和miR-2116-3p抑制物组,分别转染对应的序列,用CCK-8法与EdU试剂盒检测细胞增殖活...
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| Published in | 中国组织工程研究 Vol. 27; no. 28; pp. 4480 - 4486 |
|---|---|
| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
新疆医科大学第一附属医院,新疆维吾尔自治区乌鲁木齐市 830011
2023
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| Subjects | |
| Online Access | Get full text |
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| Abstract | R446%R318%R62; 背景:目前多项研究证实了miRNA影响增生性瘢痕的发生发展,Ⅰ型胶原与增生性瘢痕成纤维细胞密切相关,猜测miR-2116-3p也有可能与增生性瘢痕的发生发展有关系.目的:探讨miR-2116-3p在人增生性瘢痕中的表达及作用.方法:收集新疆医科大学第一附属医院6例患者的增生性瘢痕组织与6例患者重睑术后正常皮肤组织,采用qRT-PCR法检测miR-2116-3p与Ⅰ型胶原的表达.取增生性瘢痕组织,原代培养第3-6代成纤维细胞,分为阴性对照组、miR-2116-3p模拟物组和miR-2116-3p抑制物组,分别转染对应的序列,用CCK-8法与EdU试剂盒检测细胞增殖活力,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡,qRT-PCR法与Western blot法检测Ⅰ型胶原、Ⅲ型胶原和α平滑肌肌动蛋白的基因与蛋白表达,双荧光素酶实验验证其靶向结合.结果 与结论:①增生性瘢痕组织中miR-2116-3p的mRNA表达量低于正常皮肤组织(P<0.01),Ⅰ型胶原的mRNA表达量低于正常皮肤组织(P<0.01);②转染后24,48,72 h,与阴性对照组比较,miR-2116-3p模拟物组细胞活力降低(P<0.05或P<0.01),miR-2116-3p抑制物组细胞活力升高(P<0.05或P<0.01);转染后48 h,与阴性对照组比较,miR-2116-3p模拟物组细胞EdU标记数减少(P<0.01),miR-2116-3p抑制物组EdU标记数增加(P<0.05);③转染后24 h,与阴性对照组比较,miR-2116-3p模拟物组细胞划痕面积、细胞凋亡率增加(P<0.01),miR-2116-3p抑制物组细胞划痕面积、细胞凋亡率减少(P<0.01或P<0.05);④转染后24 h,与阴性对照组比较,miR-2116-3p模拟物组Ⅰ型胶原、Ⅲ型胶原和α-平滑肌肌动蛋白的基因与蛋白表达下降(P<0.05或P<0.01),miR-2116-3p抑制物组Ⅰ型胶原、Ⅲ型胶原和α-平滑肌肌动蛋白的蛋白表达升高(P<0.01);⑤双荧光素酶实验显示,miR-2116-3p能够与Ⅰ型胶原靶向结合;⑥结果表明,人增生性瘢痕中miR-2116-3p表达下调,miR-2116-3p可能通过靶向抑制Ⅰ型胶原的表达抑制人增生性瘢痕成纤维细胞的增殖、迁移,促进其凋亡. |
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| AbstractList | R446%R318%R62; 背景:目前多项研究证实了miRNA影响增生性瘢痕的发生发展,Ⅰ型胶原与增生性瘢痕成纤维细胞密切相关,猜测miR-2116-3p也有可能与增生性瘢痕的发生发展有关系.目的:探讨miR-2116-3p在人增生性瘢痕中的表达及作用.方法:收集新疆医科大学第一附属医院6例患者的增生性瘢痕组织与6例患者重睑术后正常皮肤组织,采用qRT-PCR法检测miR-2116-3p与Ⅰ型胶原的表达.取增生性瘢痕组织,原代培养第3-6代成纤维细胞,分为阴性对照组、miR-2116-3p模拟物组和miR-2116-3p抑制物组,分别转染对应的序列,用CCK-8法与EdU试剂盒检测细胞增殖活力,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡,qRT-PCR法与Western blot法检测Ⅰ型胶原、Ⅲ型胶原和α平滑肌肌动蛋白的基因与蛋白表达,双荧光素酶实验验证其靶向结合.结果 与结论:①增生性瘢痕组织中miR-2116-3p的mRNA表达量低于正常皮肤组织(P<0.01),Ⅰ型胶原的mRNA表达量低于正常皮肤组织(P<0.01);②转染后24,48,72 h,与阴性对照组比较,miR-2116-3p模拟物组细胞活力降低(P<0.05或P<0.01),miR-2116-3p抑制物组细胞活力升高(P<0.05或P<0.01);转染后48 h,与阴性对照组比较,miR-2116-3p模拟物组细胞EdU标记数减少(P<0.01),miR-2116-3p抑制物组EdU标记数增加(P<0.05);③转染后24 h,与阴性对照组比较,miR-2116-3p模拟物组细胞划痕面积、细胞凋亡率增加(P<0.01),miR-2116-3p抑制物组细胞划痕面积、细胞凋亡率减少(P<0.01或P<0.05);④转染后24 h,与阴性对照组比较,miR-2116-3p模拟物组Ⅰ型胶原、Ⅲ型胶原和α-平滑肌肌动蛋白的基因与蛋白表达下降(P<0.05或P<0.01),miR-2116-3p抑制物组Ⅰ型胶原、Ⅲ型胶原和α-平滑肌肌动蛋白的蛋白表达升高(P<0.01);⑤双荧光素酶实验显示,miR-2116-3p能够与Ⅰ型胶原靶向结合;⑥结果表明,人增生性瘢痕中miR-2116-3p表达下调,miR-2116-3p可能通过靶向抑制Ⅰ型胶原的表达抑制人增生性瘢痕成纤维细胞的增殖、迁移,促进其凋亡. |
| Author | 李培培 艾江 田文融 余扬 齐郁松 卜盼盼 马志伟 马少林 左俊 赵皎均 |
| AuthorAffiliation | 新疆医科大学第一附属医院,新疆维吾尔自治区乌鲁木齐市 830011 |
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| Author_FL | Ma Shaolin Ma Zhiwei Zuo Jun Zhao Jiaojun Qi Yusong Ai Jiang Tian Wenrong Yu Yang Li Peipei Bu Panpan |
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| Author_xml | – sequence: 1 fullname: 田文融 – sequence: 2 fullname: 左俊 – sequence: 3 fullname: 余扬 – sequence: 4 fullname: 齐郁松 – sequence: 5 fullname: 艾江 – sequence: 6 fullname: 卜盼盼 – sequence: 7 fullname: 赵皎均 – sequence: 8 fullname: 马志伟 – sequence: 9 fullname: 李培培 – sequence: 10 fullname: 马少林 |
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| Keywords | 增生性瘢痕 成纤维细胞 型胶原 皮肤纤维化 miR-2116-3p |
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| Title | miR-2116-3p在人增生性瘢痕中的表达及作用 |
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