中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (1): 59-63.doi: 10.12307/2022.010

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

miR-210/YWHAG分子轴与自体脂肪移植后促进脂肪细胞增殖及血管形成

姚炎燚,陈晓玲,张  敏,王  宏,郝立瀛,时  超,赵冰封,靳雅乔,董  玮   

  1. 包头市第八医院医疗美容科,内蒙古自治区包头市  014000
  • 收稿日期:2020-07-16 修回日期:2020-07-18 接受日期:2020-09-19 出版日期:2022-01-08 发布日期:2021-10-23
  • 通讯作者: 董玮,硕士,医师,包头市第八医院医疗美容科,内蒙古自治区包头市 014000
  • 作者简介:姚炎燚,男,1967年生,硕士,主任医师,主要从事脂肪干细胞抗衰老的研究。

miR-210/YWHAG axis with adipocyte proliferation and angiogenesis after autologous fat transplantation

Yao Yanyi, Chen Xiaoling, Zhang Min, Wang Hong, Hao Liying, Shi Chao, Zhao Bingfeng, Jin Yaqiao, Dong Wei   

  1. Department of Medical Aesthetics, Baotou Eighth Hospital, Baotou 014000, Inner Mongolia Autonomous Region, China
  • Received:2020-07-16 Revised:2020-07-18 Accepted:2020-09-19 Online:2022-01-08 Published:2021-10-23
  • Contact: Dong Wei, Master, Physician, Department of Medical Aesthetics, Baotou Eighth Hospital, Baotou 014000, Inner Mongolia Autonomous Region, China
  • About author:Yao Yanyi, Master, Chief physician, Department of Medical Aesthetics, Baotou Eighth Hospital, Baotou 014000, Inner Mongolia Autonomous Region, China

摘要:

文题释义:

血管内皮生长因子A:在血管生成、血管发生和内皮细胞生长中具有关键的调控作用,能够促进内皮细胞增殖和迁移,抑制细胞凋亡并诱导血管透化。
内皮抑素:是目前作用最强、实验效果最好的血管生成抑制剂,其能够促进血管内皮细胞增殖和迁移,诱导内皮细胞凋亡。

背景:自体脂肪组织因其无免疫排斥反应、取材便利、操作便捷等优点己成为一种优良的填充材料。研究发现,移植后的脂肪细胞在血管网重建之前一直处于缺血缺氧环境中,此类环境极易使脂肪细胞发生凋亡坏死。如何加速脂肪细胞增殖和血管形成过程且抑制其凋亡是目前提高自体脂肪移植存活的重点。
目的:探讨miR-210通过YWHAG对自体脂肪移植成活的作用机制。
方法:剥离背部自体脂肪移植8周后SD大鼠成活脂肪组织,RT-qPCR检测脂肪组织中多个miRNAs的表达水平;双荧光素酶报告基因验证
miR-210和YWHAG的靶向关系;从脂肪组织中分离脂肪细胞,分为miR-210抑制组、YWHAG过表达组、miR-210和YWHAG同时过表达组进行细胞转染,Western blot检测血管内皮生长因子、Endostatin及YWHAG的表达水平,CCK-8、Annexin V-FITC/PI及血管形成实验检测miR-210/YWHAG分子轴对脂肪细胞增殖、凋亡及血管生成的影响。

结果与结论:①miR-210在脂肪组织中高表达(P < 0.001);②双荧光素酶报告基因结果表明,YWHAG是miR-210靶向蛋白(P < 0.05);③敲降miR-210可显著抑制脂肪细胞增殖和血管生成并诱导其凋亡(P < 0.05);④同时过表达miR-210和YWHAG对脂肪细胞增殖、凋亡和血管生成则无显著影响;⑤实验表明,miR-210通过靶向下调YWHAG促进脂肪细胞增殖和血管生成并抑制其凋亡,进而促进自体脂肪移植成活。

https://orcid.org/0000-0003-1131-7268(姚炎燚) 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 自体脂肪移植, miR-210, YWHAG, 细胞增殖, 血管生成, 细胞凋亡

Abstract: BACKGROUND: Autologous adipose tissue had been became an excellent filling material because of its advantages of no immune rejection, convenient material extraction, and convenient operation. The study found that the transplanted adipocyte had been in the environment of ischemia and hypoxia before the reconstruction of the vascular network, and such an environment was easy to induce apoptosis and necrosis of adipocytes. How to accelerate the process of proliferation and angiogenesis and inhibit the apoptosis of fat cells is the focus of research on improving the survival rate of autologous fat transplantation.
OBJECTIVE: To explore the mechanism of the effect of miR-210 on the survival of autologous fat transplantation via YWHAG.
METHODS:  After 8 weeks of autologous adipose tissue transplantation in Sprague-Dawley rats, the adipose tissue on the back was stripped. The expression levels of miRNAs in adipose tissue were detected by RT-qPCR. The targeting relationship between miR-210 and YWHAG was validated by dual luciferase reporter gene. Adipocytes were isolated from the fat tissues and divided into miR-210 inhibition group, YWHAG overexpression group, miR-210 and YWHAG overexpression group for cell transfection. The expression levels of vascular endothelial growth factor, Endostatin and YWHAG were detected by western blotting. The effects of the miR-210/YWHAG axis on proliferation, apoptosis and angiogenesis of adipocytes were detected by CCK-8, Annexin V-FITC/PI and angiogenesis assay, respectively.  
RESULTS AND CONCLUSION: (1) miR-210 was highly expressed in adipose tissue (P < 0.001). (2) The results of dual luciferase reporter gene indicated that YWHAG was targeting protein of miR-210 (P < 0.05). (3) Knockdown of miR-210 significantly inhibited proliferation, angiogenesis and induced apoptosis of adipocytes (P < 0.05). (4) Overexpression of miR-210 and YWHAG at the same time had no significant effect on adipocyte proliferation, apoptosis and angiogenesis. (5) In summary, miR-210 promoted autologous fat graft survival by downregulating the expression level of YWHAG to promote proliferation, angiogenesis and inhibiting apoptosis of adipocytes.

Key words: autologous fat transplantation, mir-210, YWHAG, cell proliferation, angiogenesis, apoptosis

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