OIP5-AS1 regulates ADSCs-Exos secretion and promotes wound healing through VEGF165

doi:10.3760/cma.j.cn441417-20240904-02014

Abstract

Abstract:

Objective To investigate the mechanism of exosomes derived from ADSCs (ADSCs-Exos) knocking down opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) in wound healing. Methods　The adipose-derived stem cells (ADSCs) were isolated from the human normal subcutaneous adipose tissue of 10 patients taking selective liposuction or plastic surgery at Yan'an University Affiliated Hospital from February to June 2024. Subsequently, the exosomes derived from ADSCs supernatant were extracted, and the expressions of exosome marker proteins tumor susceptibility gene 101(TSG101) and CD9 were identified. The expression of OIP5-AS1 was determined using quantitative real-time polymerase chain reaction. The HaCaT cells were treated with H₂O₂ to construct an in vitro skin injury model. MTT assay and flow cytometry were used to detect the cell viability and apoptosis. t test and variance analysis were used for the statistical comparisons. Results　ADSCs-Exos increased the viability and inhibited the apoptosis of the H₂O₂-treated HaCaT cells (t=4.358 and 5.654; both P0.05). OIP5-AS1 expression was significantly upregulated in ADSCs-Exos as compared with ADSCs (t=4.125, P0.01), and the knockdown of OIP5-AS1 inhibited the reparative effect of ADSCs-Exos on wound healing (t=3.367 and 6.674; both P0.05). The knockdown of OIP5-AS1 inhibited the expression of vascular endothelial growth factor 165 (VEGF165) (t=3.105; P0.01), and the overexpression of VEGF165 partially reversed the inhibitory effect of the knockdown of OIP5-AS1 on wound healing (t=3.327 and 5.544; both P0.05). Conclusion　ADSC-Exos with knockdown of OIP5-AS1 affect the wound healing process regulating the expression of VEGF165, providing a new target for the treatment of skin wound healing.

Key words:

Wound, Healing, Exosomes derived from ADSCs, Opa-interacting protein 5 antisense RNA 1, Vascular endothelial growth factor 165

摘要：

目的　探讨敲低Opa相互作用蛋白5-反义转录物1（opa-interactingprotein 5 antisense RNA 1，OIP5-AS1）的脂肪间充质干细胞外泌体（exosomes derived from ADSCs，ADSCs-Exos）在创面愈合中的作用机制。方法　选取2024年2月至6月期间在延安大学附属医院接受选择性吸脂或手术整形的10例人体正常皮下脂肪组织，从中分离脂肪间充质干细胞（adipose-derived stem cell，ADSCs），提取外泌体，并鉴定外泌体标志蛋白肿瘤易感基因101（tumor susceptibility gene 101，TSG101）和CD9的表达。使用实时荧光定量聚合酶链式反应测定OIP5-AS1的表达。采用H₂O₂处理人永生化表皮细胞HaCaT，构建体外皮肤损伤模型。MTT法和流式细胞术检测细胞活力和凋亡。采用t检验、方差分析进行统计比较。结果　ADSCs-Exos增加H₂O₂处理的HaCaT细胞活力，抑制细胞凋亡（t=4.358、5.654，均P0.05）。与ADSCs相比，OIP5-AS1在ADSCs-Exos中表达显著上调（t=4.125，P0.01），且敲低OIP5-AS1抑制了ADSCs-Exos对创面愈合的修复作用（t=3.367、6.674，均P0.05）。敲低OIP5-AS1后抑制了血管内皮生长因子165（vascular endothelial growth factor 165，VEGF165）的表达（t=3.105，P0.01），VEGF165过表达可部分逆转敲低OIP5-AS1对创面愈合的抑制作用（t=3.327、5.544，均P0.05）。结论　敲低OIP5-AS1的ADSC-Exos通过调控VEGF165的表达影响创面愈合过程，为皮肤创面愈合的治疗提供新靶点。

关键词:

创面, 愈合, 脂肪间充质干细胞外泌体, Opa相互作用蛋白5-反义转录物1, 血管内皮生长因子165

Hou Guoling, Gao Dongliang, Qu Wanli, Li Ming, Wang Yakang.

OIP5-AS1 regulates ADSCs-Exos secretion and promotes wound healing through VEGF165 [J]. International Medicine and Health Guidance News, 2025, 31(2): 247-252.

侯国玲高栋梁屈万丽李明王亚康.

OIP5-AS1通过VEGF165调控ADSCs-Exos分泌促进创面愈合的研究 [J]. 国际医药卫生导报, 2025, 31(2): 247-252.

References

[1] Levoux J, Prola A, Lafuste P, et al. Platelets facilitate the wound-healing capability of mesenchymal stem cells by mitochondrial transfer and metabolic reprogramming[J]. Cell Metab, 2021,33(2):283-299.e9. DOI: 10.1016/j.cmet.2020.12.006.
[2] Takagi T, Okayama T, Asai J, et al. Topical application of sustained released-carbon monoxide promotes cutaneous wound healing in diabetic mice[J]. Biochem Pharmacol, 2022,199:115016. DOI: 10.1016/j.bcp.2022.115016.
[3] Ju C, Liu D. Exosomal microRNAs from Mesenchymal stem cells: novel therapeutic effect in wound healing[J]. Tissue Eng Regen Med, 2023,20(5):647-660. DOI: 10.1007/s13770-023-00542-z.
[4] Wang Y, Ma D, Wu Z, et al. Clinical application of mesenchymal stem cells in rheumatic diseases[J]. Stem Cell Res Ther, 2021,12(1):567. DOI: 10.1186/s13287- 021-02635-9.
[5] 刘丽桦,刘德伍. 脂肪间充质干细胞外泌体源非编码RNA在创面愈合中的作用[J]. 中国临床药理学与治疗学,2024,29(9):1049-1056. DOI:10.12092/j.issn.1009-2501. 2024.09.011.
[6] 武禄,刘标,黎彦,等. 年轻化间充质干细胞条件培养基促进小鼠创面愈合的研究[J]. 解放军医学院学报,2024,45(5):535-543. DOI:10.12435/j.issn.2095-5227.2024.047.
[7] Yang H, Li C, Li Y, et al. Adipose-derived stem cells and obesity: the spear and shield relationship[J]. Genes Dis, 2021,10(1):175-186. DOI: 10.1016/j.gendis.2021.09.004.
[8] Toh WS, Lai RC, Hui JHP, et al. MSC exosome as a cell-free MSC therapy for cartilage regeneration: implications for osteoarthritis treatment[J]. Semin Cell Dev Biol, 2017,67:56-64. DOI: 10.1016/j.semcdb.2016.11.008.
[9] Zhuang SJ, Sun XY, Luo M, et al. Hypoxic mesenchymal stem cells promote diabetic wound healing in rats by increasing VEGF secretion[J]. Biomed Environ Sci, 2023,36(8):756-759. DOI: 10.3967/bes2023.100.
[10] Qiao Z, Wang X, Zhao H, et al. The effectiveness of cell-derived exosome therapy for diabetic wound: a systematic review and meta-analysis[J]. Ageing Res Rev, 2023,85:101858. DOI: 10.1016/j.arr.2023.101858.
[11] He L, Zhu C, Jia J, et al. ADSC-Exos containing MALAT1 promotes wound healing by targeting miR-124 through activating Wnt/β-catenin pathway[J]. Biosci Rep, 2020,40(5):BSR20192549. DOI: 10.1042/BSR20192549.
[12] Heo JS, Kim S, Yang CE, et al. Human adipose mesenchymal stem cell-derived exosomes: a key player in wound healing[J]. Tissue Eng Regen Med, 2021,18(4):537-548. DOI: 10.1007/s13770-020-00316-x.
[13] Zhou K, Guo S, Tong S, et al. Immunosuppression of human adipose-derived stem cells on T cell subsets via the reduction of NF-kappaB activation mediated by PD-L1/PD-1 and Gal-9/TIM-3 pathways[J]. Stem Cells Dev, 2018,27(17):1191-1202. DOI: 10.1089/scd.2018.0033.
[14] Zhao W, Zhang H, Liu R, et al. Advances in immunomodulatory mechanisms of mesenchymal stem cells-derived exosome on immune cells in scar formation[J]. Int J Nanomedicine, 2023,18:3643-3662. DOI: 10.2147/IJN.S412717.
[15] Mazini L, Rochette L, Admou B, et al. Hopes and limits of Adipose-Derived Stem Cells (ADSCs) and Mesenchymal Stem Cells (MSCs) in wound healing[J]. Int J Mol Sci, 2020,21(4):1306. DOI: 10.3390/ijms21041306.
[16] An Y, Lin S, Tan X, et al. Exosomes from adipose-derived stem cells and application to skin wound healing[J]. Cell Prolif, 2021,54(3):e12993. DOI: 10.1111/cpr.12993.
[17] Xiao S, Xiao C, Miao Y, et al. Human acellular amniotic membrane incorporating exosomes from adipose-derived mesenchymal stem cells promotes diabetic wound healing[J]. Stem Cell Res Ther, 2021,12(1):255. DOI: 10.1186/s13287-021-02333-6.
[18] Choi EW, Seo MK, Woo EY, et al. Exosomes from human adipose-derived stem cells promote proliferation and migration of skin fibroblasts[J]. Exp Dermatol, 2018,27(10):1170-1172. DOI: 10.1111/exd.13451.
[19] Shen K, Wang XJ, Liu KT, et al. Effects of exosomes from human adipose-derived mesenchymal stem cells on inflammatory response of mouse RAW264.7 cells and wound healing of full-thickness skin defects in mice[J]. Zhonghua Shao Shang Yu Chuang Mian Xiu Fu Za Zhi, 2022,38(3):215-226. DOI: 10.3760/cma.j.cn501120-20201116-00477.
[20] Tang J, Fu C, Li Y, et al. Long noncoding RNA OIP5-AS1 promotes the disease progression in nasopharyngeal carcinoma by targeting miR-203[J]. Biomed Res Int, 2021,2021:9850928. DOI: 10.1155/2021/9850928.
[21] Xie R, Liu L, Lu X, et al. LncRNA OIP5-AS1 facilitates gastric cancer cell growth by targeting the miR-422a/ANO1 axis[J]. Acta Biochim Biophys Sin (Shanghai), 2020,52(4):430-438. DOI: 10.1093/abbs/gmaa012.
[22] Jiang W, Ou ZL, Zhu Q, et al. LncRNA OIP5-AS1 aggravates the stemness of hepatoblastoma through recruiting PTBP1 to increase the stability of β-catenin[J]. Pathol Res Pract, 2022,232:153829. DOI: 10.1016/j.prp.2022. 153829.
[23] Solovyeva VV, Chulpanova DS, Tazetdinova LG, et al. In vitro angiogenic properties of plasmid DNA encoding SDF-1α and VEGF165 genes[J]. Appl Biochem Biotechnol, 2020,190(3):773-788. DOI: 10.1007/s12010-019- 03128-5.
[24] 付文,王向臣,王延桂,等. 脂肪源性间充质干细胞外泌体在大鼠全层皮肤缺损创面愈合中的机制研究[J]. 组织工程与重建外科杂志,2023,19(4):342-351,379. DOI:10.3969/j.issn.1673-0364.2023.04.003.
[25] Salafutdinov II, Gazizov IM, Gatina DK, et al. Influence of recombinant codon-optimized plasmid DNA encoding VEGF and FGF2 on co-induction of angiogenesis[J]. Cells, 2021,10(2):432. DOI: 10.3390/cells10020432.
[26] Eggers C, Müller J, Schultze-Mosgau S. VEGF transfer based on gene-modified fibroblasts using a hypoxia-induced vector to modulate neoangiogenesis in ischaemic regions of myocutaneous transplants[J]. Int J Oral Maxillofac Surg, 2015,44(2):267-276. DOI: 10.1016/j.ijom.2014.06.018.