Role of autophagy during cutaneous wound healing in mice

Na ZHAO; Dengqun LIU; Guojian WANG; Shuang LONG; Huimin WAN; Yongping SU; Tao WANG

doi:10.16016/j.1000-5404.201808194

您当前的位置：

首页 >

文章列表页 >

Role of autophagy during cutaneous wound healing in mice

Basic Medicine | 更新时间：2026-01-09

- Role of autophagy during cutaneous wound healing in mice
- Journal of Third Military Medical University Vol. 41, Issue 1, Pages: 25-32(2019)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)军事预防医学系防原医学教研室，全军复合伤研究所，创伤、烧伤与复合伤国家重点实验室，重庆市纳米医学工程研究中心
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81372061);the General Program of National Natural Science Foundation of China(81673089);the Basic and Frontier Research of Chongqing(CSTC2016jcyjA0381)
- DOI：10.16016/j.1000-5404.201808194
  CLC： R322.99;R329.28;R641
- Received：30 August 2018，
  
  Revised：2018-10-19，
  
  Published：15 January 2019
- 稿件说明：
移动端阅览
Na ZHAO, Dengqun LIU, Guojian WANG, et al. Role of autophagy during cutaneous wound healing in mice[J]. Journal of Third Military Medical University, 2019, 41(1): 25-32.
DOI：

Na ZHAO, Dengqun LIU, Guojian WANG, et al. Role of autophagy during cutaneous wound healing in mice[J]. Journal of Third Military Medical University, 2019, 41(1): 25-32. DOI： 10.16016/j.1000-5404.201808194.

摘要

目的

观察自噬在小鼠皮肤创面愈合中的发生情况，探讨其在创面修复中的可能作用。

方法

以GFP-LC3转基因小鼠制备皮肤后背部创面，在致伤后0、3、7、14 d取皮肤创面组织制作石蜡切片进行GFP免疫荧光染色标记自噬点，观察自噬在创面愈合过程中的动态变化；采用GFP抗体与α-SMA、F4/80抗体进行荧光双标，明确肉芽间质中发生自噬的主要细胞类型。而后将GFP-LC3小鼠分为自噬抑制剂3-甲基腺嘌呤(3-MA)处理组与对照组，伤后连续10 d腹腔注射3-MA(10 mg/kg)，动态观察创面大体愈合情况，在0、7、10 d取创面皮肤，组织学固定染色，评价自噬对创面愈合的影响；同时采用定量PCR(qRT-PCR)检测α-SMA、TGF-β1、PAI-1、Ⅰ型和Ⅲ型胶原等分子的mRNA表达，评价自噬对创面修复过程中胶原与胞外基质表达水平的影响。

结果

创面愈合过程中自噬水平先增强后逐渐回落，在增殖期(7 d)的自噬水平最高；肉芽角质细胞自噬强于间质细胞，肉芽间质中发生自噬的细胞主要为α-SMA阳性肌成纤维细胞；与对照组相比，3-MA处理组在伤后7 d与10 d创面肉芽的面积、宽度与表皮厚度差异均无统计学意义，而处理组7 d表皮基底细胞增殖活力增强(

0.05)；两组α-SMA、胶原、TGF-β1与PAI-1等分子的mRNA水平差异均无统计学意义。

结论

皮肤创面愈合过程中增殖期存在高自噬水平，但抑制小鼠创面愈合过程中的自噬水平对创面愈合未见显著影响。

Abstract

Objective

To observe the dynamic changes in autophagy during cutaneous wound healing and investigate its possible contributions to wound repair.

Methods

Two skin wound of 6 mm in size were inflicted on the back of GFP-LC3 transgenic mice. In 0

7 and 14 d after skin injury

the skin samples were harvested to prepare paraffin-embedded sections

and GFP immunohistochemical (IHC) staining with anit-LC3 antibody was performed for the dynamic changes of autophagy during wound healing. Double-labeling immunofluorescence assay was utilized for identifying cell types of autophgy in granulation tissues by staining both GFP and macrophage marker F4/80 or myofibroblasts marker α-SMA. The mice were divided into skin defect group and injured mice with 3-methyladenine (3-MA

10 mg/kg

intra-peritoneal injection for 10 continuous days) treatment group. The dynamic changes of the wound were observed

and the skin tissues were collected in 0

7 and 10 d after injury to evaluate the effect of autophagy on wound healing after fixation and staining. qRT-PCR was employed to detect the expression levels of α-SMA

TGF-β1

PAI-1

type Ⅰ and Ⅲ collagen in the wound samples.

Results

The autophagic activity was enhanced and then declined in the process of wound healing

and its strongest activity was obse

rved in proliferative phase (7 d). In the granulation tissues

the activity was stronger in the keratinocytes than the interstitial cells

and the autophagic cells were mainly α-SMA-positive myofibroblasts. There were no significant differences in wound area and width

and epidermis thickness in 7 and 10 d after skin injury between the 3-MA treatment and untreated mice

but the former group had significantly increased proliferation activity of epidermal basal cells (

0.05) at 7 d after injury. However

no significant differences were seen in the mRNA levels of α-SMA

TGF-β1

PAI-1 and collagen molecules between the 2 groups of mice.

Conclusion

There exists stronger autophagic activity in the proliferative phase of cutaneous wound healing

but inhibiting autophagy shows little impact on the process.

关键词

Keywords

references

GURTNER G C, WERNER S, BARRANDON Y, et al . Wound repair and regeneration[J ] . Nature, 2008, 453(7193): 314-321. DOI:10.1038/nature07039.

MIZUSHIMA N, KOMATSU M. Autophagy: renovation of cells and tissues[J]. Cell, 2011, 147(4): 728-741. DOI:10.1016/j.cell.2011.10.026.

SARKAR S, PERLSTEIN E O, IMARISIO S, et al . Small molecules enhance autophagy and reduce toxicity in Huntington's disease models[J ] . Nat Chem Biol, 2007, 3(6): 331-338. DOI:10.1038/nchembio883.

WANG S Y, YU Q J, ZHANG R D, et al . Core signaling pathways of survival/death in autophagy-related cancer networks[J ] . Int J Biochem Cell Biol, 2011, 43(9): 1263-1266. DOI:10.1016/j.biocel.2011.05.010.

UCAR A, GUPTA S K, FIEDLER J, et al . The miRNA-212/132 family regulates both cardiac hypertrophy and cardiomyocyte autophagy[J ] . Nat Commun, 2012, 3(1): 1078-1089. DOI:10.1038/ncomms2090.

MIZUSHIMA N, YAMAMOTO A, MATSUI M, et al . In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker[J ] . Mol Biol Cell, 2004, 15(3): 1101-1111. DOI:10.1091/mbc.e03-09-0704..

WAN G, XIE W, LIU Z, et al . Hypoxia-induced MIR155 is a potent autophagy inducer by targeting multiple players in the MTOR pathway[J ] . Autophagy, 2014, 10(1): 70-79.DOI:10.4161/auto.26534.

GUO Y, LIN C, XU P, et al . AGEs induced autophagy impairs cutaneous wound healing via stimulating macrophage polarization to M1 in diabetes[J ] . Sci Rep, 2016, 6(1): 36416-36432. DOI:10.1038/srep36416..

ASAI E, YAMAMOTO M, UEDA K, et al . Spatiotemporal alterations of autophagy marker LC3 in rat skin fibroblasts during wound healing process[J ] . Fukushima J Med Sci, 2018, 64(1): 15-22. DOI: 10.5387/fms.2016-13.

KAHN D, SPEARMAN C W, MALL A, et al . The effect of rapamycin on the healing of the ureteric anastomosis and wound healing[J ] . Transplant Proc, 2005, 37(2):830-831. DOI:10.1016/j.transproceed.2004.12.156.

LIU H, CAO M M, WANG Y, et al . Endoplasmic reticulum stress is involved in the connection between inflammation and autophagy in type 2 diabetes[J ] . Gen Comp Endocrinol, 2015, 210: 124-129. DOI:10.1016/j.ygcen.2014.09.006.

BELLEUDI F, PURPURA V, CAPUTO S, et al . FGF7/KGF regulates autophagy in keratinocytes:a novel dual role in the induction of both assembly and turnover of autophagosomes[J ] .Autophagy, 2014, 10(5):803-821.DOI:10.4161/auto.28145.

BERNARD M, DIEUDÉ M, YANG B, et al . Autophagy fosters myofibroblast differentiation through MTORC2 activation and downstream upregulation of CTGF[J ] . Autophagy, 2014, 10(12):2193-2207.DOI:10.4161/15548627.2014.981786.

ZHENG W,QIAN Y, CHEN S, et al . Rapamycin protects against peritendinous fibrosis through activation of autophagy[J ] . Front Pharmacol, 2018, 9: 402. DOI:10.3389/fphar.2018.00402.

VESCARELLI E, PILLONI A, DOMINICI F, et al . Autophagy activation is required for myofibroblast differentiation during healing of oral mucosa[J ] . J Clin Periodontol, 2017, 44(10):1039-1050.DOI:10.1111/jcpe.12767.

SHI J, SHI S, WU B, et al . Autophagy protein LC3 regulates the fibrosis of hypertrophic scar by controlling Bcl-xL in dermal fibroblasts[J ] .Oncotarget, 2017, 8(55): 93757-93770. DOI:10.18632/oncotarget.20771.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of bFGF in coordination with autografting of bone marrow mesenchymal stem cells on expression of PCNA and Ⅷ-R Ag of porcine skin defect wound

Promoting effect of ultra-low dose DMSO on fibroblast proliferation and its underlying mechanism

RANKL通过ROS介导HIF-1α下调促进自噬和破骨细胞分化

转录因子EB增强巨噬细胞自噬功能及逆转内毒素耐受的作用

Autophagy inducer spermidine improves brain injury after intracerebral hemorrhage

Related Author

陈涛

付小兵

伍津津

韩冰

孙同柱

TAN Ranjing

KANG Tehao

WU Dengyan

Related Institution

Center for Dermatology and Medical Cosmetology， the Second Affiliated Hospital of Chongqing Medical University

Department of Dermatology， Plastic and Cosmetic Surgery， Wushan People’s Hospital

Department of Cerebrovascular Diseases， Second Affiliated Hospital of Zhengzhou University, Zhengzhou University

Academy of Medical Sciences, Zhengzhou University

⁰