miR-199a-5p inhibits proliferation and migration of burn-denatured fibroblasts by targeting VEGFA in burn patients
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Qinghua Wu
and
Wenna Li
Abstract
Objectives
The skin altered by burn injury has the capacity to revert to its normal function and structure. Nonetheless, the exact mechanism behind this restoration remains unclear. This study was to examine the function of miR-199a-5p in the migration and proliferation of fibroblasts affected by burn injury.
Methods
The miR-199a-5p expression was assessed using quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation and migration capacity of human skin fibroblast (HSF) cells were measured by CCK-8 and Transwell assay. The protein levels of vascular endothelial growth factor A (VEGFA), phosphorylated phosphoinositide 3-kinases (p-PI3K), phosphorylated protein kinase B (p-AKT), and phosphorylated endothelial nitric oxide synthase (p-eNOS) were assessed via western blot assay.
Results
The miR-199a-5p levels were decreased in individuals with burn injuries as well as in burn-denatured HSF cells, at the same time, there was an increase in the VEGFA mRNA levels. miR-199a-5p negatively regulated VEGFA, which in turn suppressed the migration and proliferation of HSF cells. Additionally, miR-199a-5p inhibited the PI3K/AKT/eNOS signaling pathway through the regulation of VEGFA.
Conclusions
miR-199a-5p regulates proliferation and migration of burn degenerated fibroblasts by targeting VEGFA.
Introduction
The treatment of profound second-degree or mixed-degree burn injuries represents a considerable hurdle and a primary area of investigation within the field of burn surgery [1]. Altered dermal tissue is commonly encountered within burn injuries, where it influences disrupted cellular metabolism, compromised functionality, and morphological abnormalities. Nonetheless, these alterations have the potential to be reversed, and the tissue can regain its normal structure and function once the adjacent microenvironment is optimized [2]. Emerging clinical evidence has demonstrated that prompt debridement coupled with the conservation of the dermis can enhance the rate of healing, diminish scarring, and lead to better functional results in patients with deep burn wounds [3], 4]. Maintaining the altered dermis, including the sustenance of human skin fibroblast (HSF) migration and proliferation, has the potential to enhance the scar tissue quality during the recovery from burn injuries [5], 6]. Nevertheless, the precise molecular processes by which the altered dermis contributes to wound repair remain unelucidated.
MicroRNAs (miRNAs) are essential for regulating human gene expression post-transcriptionally. This interaction can cause either degradation or the inhibition of mRNA translation [7]. Furthermore, an increasing amount of research has uncovered that numerous miRNAs are essential in the processes of wound repair, skin development, and regeneration. They exert their influence by governing the differentiation, apoptosis, and proliferation of skin cells [8], 9].
miR-199a-5p can modulate the function of skin cancer cells [10]. The suppression of miR-199a-5p markedly enhanced and notably accelerated the healing process of diabetic foot ulcers [11]. Moreover, miR-199a-5p has been pinpointed as a miRNA with dysregulated expression in keloid tissue, where it significantly influences the growth of keloid fibroblasts [12]. Currently, vascular endothelial growth factor A (VEGFA) stands out as a potent angiogenic cytokine can enhance the permeability of small veins and larger veins. It facilitates the division and growth of vascular endothelial cells, triggers angiogenesis, and is crucial in various biological processes, including tissue repair, embryonic development, cancer progression, and the spread of metastasis [13], 14]. Research has indicated that VEGFA is integral in governing the process of recovery of wounds following burn injuries [4], and miR-199a-5p was found to suppress the growth of dermal cells in cashmere goats by modulating the levels of VEGFA expression [15]. However, whether miR-199a-5p regulates VEGFA and participates in wound healing after burn remains unknown.
Materials and methods
Participants
In this study, 35 burn patients over 18 years old admitted to Chengdu Second People’s Hospital were included, the average age of these patients was 34 ± 8 years old. In addition, there were 20 cases of burn degree II and 15 cases of burn degree III. As reported in prior studies [3], the specimens were obtained through tangential excision of eschar and large sheets of split-thickness auto graft with preservation of dermis on the fourth day following the occurrence of the burn injury. The adjacent unburned skin tissues were utilized as the control group. Approval was obtained from the ethics committee of Chengdu Second People’s Hospital (approve number: 2023031, date: 06-03-2023). Written informed consent was acquired by each subject.
Cell culture and thermal injury
HSF cell lines were cultivated at 37 °C in Dulbecco’s modified eagle medium (DMEM) with 10 % fetal bovine serum (FBS). Upon achieving a cell density of 50–60 %, the cells were placed in a water bath set to 52 °C for a brief period of 30 s. The HSF suspension in control group was subjected to a 30-s incubation period in water maintained at 37 °C.
Cell transfection
miR-199a-5p inhibitor, miR-199a-5p mimic, negative control, VEGFA overexpression vector (oe-VEGFA) and empty vector (oe-NC) were synthesized by GenePharma (Shanghai, China). Transfection was performed using Lipofectamine 3,000 (Thermo Fisher Scientific, Waltham, MA, USA).
Quantitative real-time polymerase chain reaction (qRT-PCR)
The total RNA was isolated using Trizol (Invitrogen, Carlsbad, CA, USA), and the mirVana miRNA Isolation Kit (Ambion, Grand Island, NY, USA) was employed to purify miRNAs. Subsequently, the levels of VEGFA mRNA and miR-199a-5p were quantified utilizing the One-Step RT-PCR Kit (TaKaRa, Otsu, Japan). qRT-PCR was performed using the CFX96 Touch qRT-PCR System (Bio-Rad, Hercules, CA, USA). Relative expression was conducted using 2–ΔΔCt method, with β-actin serving as the normalization control.
CCK-8 assay
CCK-8 kit (Dojindo, Kumamoto, Japan) was employed to assess the capacity for cell proliferation. Following transfection, HSF cells were distributed with each well containing 5,000 cells into 96-well plates in triplicate, CCK-8 reagent was introduced into each well at intervals of 24 h. After a 2-h incubation period, the optical density at a wavelength of 450 nm was recorded.
Transwell assay
Cells in serum-free DMEM were introduced into the up chamber. After that, DMEM containing 10 % FBS was added to the lower chamber. After a 24-h incubation period, the cells that had adhered to the membrane’s upper surface were carefully removed by a cotton swab. Subsequently, the cells were fixed for 30 min using paraformaldehyde and then stained with crystalline violet for 15 min. The cell number was counted under an optical microscope.
Luciferase reporter assay
We searched for the downstream target genes of miR-199a-5p using the starBase database and predicted the binding sites of miR-199a-5p and VEGFA by using starBase online site. The wild-type and mutant 3′-UTR of VEGFA were constructed and inserted into the pmirGLO vector. The plasmids were constructed and co-transfected into the cells along with control mimics/inhibitors or miR-199a-5p mimics/inhibitors by utilizing Lipofectamine 3,000. Following a 48-h transfection period, the Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA) was used to measure luciferase activity.
Western blot
RIPA buffer (Beyotime Biotechnology, Nantong, China) was used for cell lysis. Protein concentration was measured using the bicinchoninic acid kit (Tiangen, Beijing, China). Protein samples, consisting of 20 µg each, were separated via SDS-PAGE and subsequently transferred to PVDF membranes for immunoblotting. The membrane was incubated overnight with primary antibodies against VEGFA (ab52917), phosphorylated phosphoinositide 3-kinases (p-PI3K) (ab302958), phosphorylated protein kinase B (p-AKT) (ab8805), and phosphorylated endothelial nitric oxide synthase (p-eNOS) (ab215717) (Abcam, Cambridge, MA, USA). The membranes were then incubated for 2 h at room temperature with the matching HRP-conjugated secondary antibody. An ECL kit (Millipore, Bedford, MA, USA) was used to visualize the protein bands on a ChemiDoc XRS imaging system, and the resulting blots were analyzed with ImageJ software.
Statistical analysis
To assess the differences among groups, statistical comparisons between two groups were conducted using the Student’s t-test, while multi-group analyses were performed through one-way or two-way ANOVA. To elucidate the intricate relationship between miR-199a-5p and VEGFA, Pearson correlation analysis was employed to meticulously examine their association. Statistical evaluations were conducted using SPSS 26.0 (IBM SPSS Statistics for Windows, NY, USA) and GraphPad Prism 9.0 (GraphPad Prism software, CA, USA) software. Statistical significance has been defined as a p-value of <0.05.
Results
Expression of miR-199a-5p and VEGFA in burn wound tissues
A significant reduction in the level of miR-199a-5p was observed 4 days following thermal injury (Figure 1A). Conversely, the expression of VEGFA was elevated in tissues 4 days post-burn injury (Figure 1B). Furthermore, the levels of VEGFA and miR-199a-5p showed a substantial inverse connection (Figure 1C).
Expression of miR-199a-5p and VEGFA in burn wound tissues. (A) Expression of miR-199a-5p in normal tissues (n=35) and burn tissues (n=35). (B) Expression of VEGFA in normal tissues (n=35) and burn tissues (n=35). (C) Correlation between expression levels of miR-199a-5p and VEGFA in burn tissues. ***p<0.001.
Role of miR-199a-5p overexpression and inhibition in burn-denatured fibroblasts
In this study, we initially developed a thermal injury model, subsequent to which we assessed the levels of miR-199a-5p in HSF cells subjected to heat damage. The expression of miR-199a-5p decreased in a time-dependent manner (6, 12, 24, 48 h) after thermal injury (Figure 2A). The expression of miR-199a-5p was elevated in the cells treated with the miR-199a-5p mimic, whereas it was reduced in those treated with the inhibitor (Figure 2B). miR-199a-5p overexpression decreased the proliferative capacity of cells subjected to thermal damage. The suppression of miR-199a-5p resulted in an enhancement of the proliferative capacity in these cells (Figure 2C). The cell migration capacity was observed to diminish after upregulation of miR-199a-5p. Conversely, following the inhibition of miR-199a-5p, there was an observed improvement in the cell migration ability (Figure 2D).
Effects of miR-199a-5p overexpression and inhibition in burn-denatured fibroblasts. (A) Expression of miR-199a-5p in heat-denatured HSF cells. (B) miR-199a-5p was overexpressed or inhibited in heat-denatured HSF cells. (C) Changes in proliferation of heat-denatured HSF cells after overexpression or inhibition of miR-199a-5p. (D) Changes in migration ability of heat-denatured HSF cells after overexpression or inhibition of miR-199a-5p. The experiment was repeated three times (n=3). ***p<0.001, **p<0.01, *p<0.05.
VEGFA is a target of miR-199a-5p
Time-dependent increase in VEGFA expression was observed at different time points after thermal injury (Figure 3A). Figure 3B illustrates the interaction sites and the binding relationships between miR-199a-5p and VEGFA. Elevating the levels of miR-199a-5p resulted in a reduction of luciferase activity associated with VEGFA. In contrast, the suppression of miR-199a-5p led to an elevation in the luciferase activity of VEGFA. miR-199a-5p overexpression decreased the expression level of VEGFA, and transfection of VEGFA overexpression vector reversed the impact of miR-199a-5p on VEGFA level, but oe-VEGFA had no effect on miR-199a-5p (Figure 3C and D).
VEGFA is a target of miR-199a-5p. (A) Expression of VEGFA in heat-denatured HSF cells. (B) The relationship between VEGFA and miR-199a-5p was verified via dual-luciferase reporting experiments. (C, D) miR-199a-5p could regulate the expression of VEGFA. The experiment was repeated three times (n=3). ***p<0.001, **p<0.01, ns, no significant difference.
miR-199a-5p inhibits the PI3K/AKT/eNOS signaling pathway by targeting VEGFA.
Elevated levels of VEGFA could counter the suppression of cell proliferation and migration that is triggered by the upregulation of miR-199a-5p (Figure 4A and B). Furthermore, miR-199a-5p overexpression resulted in decrease of VEGFA, p-AKT, p-PI3K, and p-eNOS protein expression levels. Nevertheless, this effect was reversed following the overexpression of VEGFA (Figure 4C).
miR-199a-5p inhibits the PI3K/AKT/eNOS signaling pathway by targeting VEGFA. (A, B) miR-199a-5p regulates the proliferation and migration of burn-denatured fibroblasts by targeting VEGFA. (C) Expression of VEGFA, p-PI3K, p-AKT, and p-eNOS in heat-denatured HSF cells. The experiment was repeated three times (n=3). ***p<0.001.
Discussion
Recently, extensive research has been conducted on the cellular mechanisms involved in the wound repair process. The activation of specific signaling pathways is crucial for the proper progression of wound healing [16]. The migration and growth of HSF are pivotal in the wound repair sequence initiated by burn injuries [17]. The findings of this investigation indicated that heat stress markedly reduces miR-199a-5p level and concurrently elevates the levels of VEGFA. Furthermore, the upregulation of miR-199a-5p was observed to lower VEGFA levels, and it also suppressed the migration and proliferation of HSF cells. In addition, miR-199a-5p regulated PI3K/AKT/eNOS signaling pathway.
miR-199a-5p is a significant player in numerous pathological conditions. For instance, earlier research has suggested that miR-199a-5p could serve as a potential biomarker for an array of cancer types [18], 19]. Furthermore, miR-199a-5p is intimately linked with atherosclerosis [20], the differentiation of osteoblasts [21], pre-eclampsia [22], as well as myocardial infarction [23]. Previous research has indicated that fibroblasts can be stimulated through heat-induced denaturation, leading to the activation of these cells, which in turn promotes the synthesis of collagen fibers and the upregulation of various growth factors [24].
miR-199a-5p expression was down-regulated in burn patients and heat-denatured HSF cells, and the cell migration and proliferation were decreased by miR-199a-5p overexpression, but they were enhanced by miR-199a-5p inhibition. The findings align with prior research [12], yet it is noteworthy that miR-199a-5p tends to exhibit an increased expression during the early phase following thermal damage, with a subsequent decline in its expression levels as time progresses. As down-regulated miR-199a-5p promotes scar formation [12], this phenomenon may be caused by the fact that fibroblasts impair cell function at the initial stage of thermal injury, resulting in up-regulated miR-199a-5p expression. As time goes on, the self-repair process is initiated, thus contributing to wound healing. The findings imply that miR-199a-5p potentially assumes a regulatory function in the behavior of human dermal fibroblasts throughout the wound repair process.
This study discovered that miR-199a-5p targets the gene VEGFA, which also validates previous studies [11]. In addition to its role in diabetic ulcerated feet [11], miR-199a-5p also regulates tumorgenesis and angiogenesis by targeting VEGFA [25] and is involved in the pathogenesis of endometriosis [26]. Numerous investigations have substantiated the angiogenic promotion by VEGFA in the context of wound repair [27], 28]. This research demonstrated that the upregulation of VEGFA counteracted the reduction in cell migration and proliferation induced by miR-199a-5p overexpression and enhanced the restorative capabilities of heat-denatured HSF cells.
Nitric oxide (NO) is an endothelium-derived vascular active compound that is pivotal in preserving vascular equilibrium [29]. eNOS is responsible for producing NO. NO synthesis is increased when the PI3K/AKT pathway is stimulated because this causes eNOS to be directly phosphorylated. Conversely, the suppression of the PI3K/AKT pathway is linked to a decrease in eNOS phosphorylation and a subsequent inhibition of NO generation, a phenomenon that correlates with impaired endothelial cell function [30]. The PI3K/AKT/eNOS signaling pathway has been implicated in endothelial cells [31] and angiogenesis [32] in previous research. The engagement of the PI3K/AKT/eNOS signaling pathway is a vital mechanism in the induction of angiogenesis [33]. More importantly, several studies have found that this pathway is involved in regulating the wound healing [33], 34]. Meanwhile, this study also found that miR-199a-5p not only inhibited the protein expression of VEGFA but also inhibited the protein levels of p-PI3K, p-AKT, and p-eNOS, and the overexpression of VEGFA reversed this trend. This may indicate that miR-199a-5p regulates burn-denatured fibroblasts by targeting VEGFA via regulating PI3K/AKT/eNOS signaling pathway.
Nonetheless, the research presented here has its limitations. For instance, this study only measured the levels of miR-199a-5p and VEGFA in the skin tissue of patients 4 days post-burn. Monitoring their expression at the onset of burn injury could render the experimental data more comprehensive. Additionally, VEGFA is merely one of the numerous target genes of miR-199a-5p, which may participate in the mechanism of wound healing post-burn through the regulation of other genes. Consequently, we intend to investigate and validate additional target genes of miR-199a-5p in future experiments, thereby providing a broader spectrum of therapeutic strategies for wound healing after burns. Furthermore, it is plausible that miR-199a-5p may also modulate the wound healing process following a burn via alternative pathways, a hypothesis that necessitates further experimental validation.
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Research ethics: Approval was obtained from the ethics committee of Chengdu Second People’s Hospital (approve number: 2023031, date: 06-03-2023). The procedures used in this study adhere to the tenets of the Declaration of Helsinki.
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Informed consent: All participants provided written informed consent.
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Author contributions: QHW designed the study, performed the experiment, and was a major contributor in writing the manuscript. WNL involved in interpretation of data, drafting the manuscript and revising it critically for important intellectual content. All authors read and approved the final manuscript.
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Use of Large Language Models, AI and Machine Learning Tools: Not applicable.
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Conflict of interest: The author states no conflict of interest.
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Research funding: This work was supported by Health Commission of Chengdu Foundation Project [grant numbers: 2023105]. Sichuan Natural Science Foundation Project [grant numbers: 2023NSFSC0550]. Science and Technology Department of Shaanxi Province - Youth project [grant numbers: 2024JC-YBQN-0946].
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Data availability: The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
References
1. Markiewicz-Gospodarek, A, Kozioł, M, Tobiasz, M, Baj, J, Radzikowska-Büchner, E, Przekora, A. Burn wound healing: clinical complications, medical care, treatment, and dressing types: the current state of knowledge for clinical practice. Int J Environ Res Publ Health 2022;19:1338. https://doi.org/10.3390/ijerph19031338.Search in Google Scholar PubMed PubMed Central
2. Yang, XH, Huang, XY, Lei, SR, Zhang, PH, Zhang, MH, Xiao, MZ, et al.. Long-term result of repair of deeply burned hands with large sheet of split-thickness autoskin grafting with the preservation of denatured dermis. Zhonghua Shaoshang Zazhi 2005;21:27–9.Search in Google Scholar
3. Guo, L, Huang, X, Liang, P, Zhang, P, Zhang, M, Ren, L, et al.. Role of XIST/miR-29a/LIN28A pathway in denatured dermis and human skin fibroblasts (HSFs) after thermal injury. J Cell Biochem 2018;119:1463–74. https://doi.org/10.1002/jcb.26307.Search in Google Scholar PubMed
4. Huang, M, Huang, X, Jiang, B, Zhang, P, Guo, L, Cui, X, et al.. linc00174-EZH2-ZNF24/Runx1-VEGFA regulatory mechanism modulates post-burn wound healing. Mol Ther Nucleic Acids 2020;21:824–36. https://doi.org/10.1016/j.omtn.2020.07.010.Search in Google Scholar PubMed PubMed Central
5. Fusté, NP, Guasch, M, Guillen, P, Anerillas, C, Cemeli, T, Pedraza, N, et al.. Barley β-glucan accelerates wound healing by favoring migration versus proliferation of human dermal fibroblasts. Carbohydr Polym 2019;210:389–98. https://doi.org/10.1016/j.carbpol.2019.01.090.Search in Google Scholar PubMed
6. Jiang, B, Li, Y, Liang, P, Liu, Y, Huang, X, Tong, Z, et al.. Nucleolin enhances the proliferation and migration of heat-denatured human dermal fibroblasts. Wound Repair Regen 2015;23:807–18. https://doi.org/10.1111/wrr.12339.Search in Google Scholar PubMed
7. Bartel, DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004;116:281–97. https://doi.org/10.1016/s0092-8674(04)00045-5.Search in Google Scholar PubMed
8. Singhvi, G, Manchanda, P, Krishna Rapalli, V, Kumar Dubey, S, Gupta, G, Dua, K. MicroRNAs as biological regulators in skin disorders. Biomed Pharmacother 2018;108:996–1004. https://doi.org/10.1016/j.biopha.2018.09.090.Search in Google Scholar PubMed
9. Cui, X, Huang, X, Huang, M, Zhou, S, Le, G, Yu, W, et al.. miR-24-3p obstructs the proliferation and migration of human skin fibroblasts after thermal injury by targeting PPAR-β and positively regulated by NF-κB. Exp Dermatol 2022;31:841–53. https://doi.org/10.1111/exd.14517.Search in Google Scholar PubMed
10. Liu, M, Wang, H, Liu, Z, Liu, G, Wang, W, Li, X. Exosomes from adipose-derived stem cells inhibits skin cancer progression via miR-199a-5p/SOX4. Biotechnol Genet Eng Rev 2024;40:3950–62. https://doi.org/10.1080/02648725.2023.2204702.Search in Google Scholar PubMed
11. Wang, H, Wang, X, Liu, X, Zhou, J, Yang, Q, Chai, B, et al.. miR-199a-5p plays a pivotal role on wound healing via suppressing VEGFA and ROCK1 in diabetic ulcer foot. Oxid Med Cell Longev 2022;2022:4791059. https://doi.org/10.1155/2022/4791059.Search in Google Scholar PubMed PubMed Central
12. Wu, ZY, Lu, L, Liang, J, Guo, XR, Zhang, PH, Luo, SJ. Keloid microRNA expression analysis and the influence of miR-199a-5p on the proliferation of keloid fibroblasts. Genet Mol Res 2014;13:2727–38. https://doi.org/10.4238/2014.april.14.2.Search in Google Scholar PubMed
13. Peach, CJ, Mignone, VW, Arruda, MA, Alcobia, DC, Hill, SJ, Kilpatrick, LE, et al.. Molecular pharmacology of VEGF-A isoforms: binding and signalling at VEGFR2. Int J Mol Sci 2018;19:1264. https://doi.org/10.3390/ijms19041264.Search in Google Scholar PubMed PubMed Central
14. Frezzetti, D, Gallo, M, Maiello, MR, D’Alessio, A, Esposito, C, Chicchinelli, N, et al.. VEGF as a potential target in lung cancer. Expert Opin Ther Targets 2017;21:959–66. https://doi.org/10.1080/14728222.2017.1371137.Search in Google Scholar PubMed
15. Zhang, T, Zheng, Y, Zhang, F, Wang, X, Du, J, Wang, X. MiR-199a-5p inhibits dermal papilla cells proliferation by regulating VEGFA expression in cashmere goat. Gene 2024;893:147901. https://doi.org/10.1016/j.gene.2023.147901.Search in Google Scholar PubMed
16. Pi, L, Fang, B, Meng, X, Qian, L. LncRNA XIST accelerates burn wound healing by promoting M2 macrophage polarization through targeting IL-33 via miR-19b. Cell Death Discov 2022;8:220. https://doi.org/10.1038/s41420-022-00990-x.Search in Google Scholar PubMed PubMed Central
17. Li, J, Chen, J, Kirsner, R. Pathophysiology of acute wound healing. Clin Dermatol 2007;25:9–18. https://doi.org/10.1016/j.clindermatol.2006.09.007.Search in Google Scholar PubMed
18. Xu, M, Zhang, J, Lu, X, Liu, F, Shi, S, Deng, X. MiR-199a-5p-regulated SMARCA4 promotes oral squamous cell carcinoma tumorigenesis. Int J Mol Sci 2023;24:4756. https://doi.org/10.3390/ijms24054756.Search in Google Scholar PubMed PubMed Central
19. Zhao, DY, Zhou, L, Yin, TF, Zhou, YC, Zhou, GY, Wang, QQ, et al.. Circulating miR-627-5p and miR-199a-5p are promising diagnostic biomarkers of colorectal neoplasia. World J Clin Cases 2022;10:5165–84. https://doi.org/10.12998/wjcc.v10.i16.5165.Search in Google Scholar PubMed PubMed Central
20. Liang, W, Chen, J, Zheng, H, Lin, A, Li, J, Wu, W, et al.. MiR-199a-5p-containing macrophage-derived extracellular vesicles inhibit SMARCA4 and alleviate atherosclerosis by reducing endothelial cell pyroptosis. Cell Biol Toxicol 2023;39:591–605. https://doi.org/10.1007/s10565-022-09732-2.Search in Google Scholar PubMed
21. Hu, J, Huang, X, Zheng, L, Zhang, Y, Zeng, H, Nie, L, et al.. MiR-199a-5P promotes osteogenic differentiation of human stem cells from apical papilla via targeting IFIT2 in apical periodontitis. Front Immunol 2023;14:1149339. https://doi.org/10.3389/fimmu.2023.1149339.Search in Google Scholar PubMed PubMed Central
22. Chen, Z, Wu, M, Huang, H, Tao, H, Zou, L, Luo, Q. Plasma exosomal miR-199a-5p derived from preeclampsia with severe features impairs endothelial cell function via targeting SIRT1. Reprod Sci 2022;29:3413–24. https://doi.org/10.1007/s43032-022-00977-0.Search in Google Scholar PubMed
23. Chen, HY, Lu, J, Wang, ZK, Yang, J, Ling, X, Zhu, P, et al.. Hsa-miR-199a-5p protect cell injury in hypoxia induces myocardial cells via targeting HIF1α. Mol Biotechnol 2022;64:482–92. https://doi.org/10.1007/s12033-021-00423-7.Search in Google Scholar PubMed
24. Martin, P. Wound healing--aiming for perfect skin regeneration. Science 1997;276:75–81. https://doi.org/10.1126/science.276.5309.75.Search in Google Scholar PubMed
25. Zhang, L, Cao, H, Gu, G, Hou, D, You, Y, Li, X, et al.. Exosomal MiR-199a-5p inhibits tumorigenesis and angiogenesis by targeting VEGFA in osteosarcoma. Front Oncol 2022;12:884559. https://doi.org/10.3389/fonc.2022.884559.Search in Google Scholar PubMed PubMed Central
26. Hsu, CY, Hsieh, TH, Tsai, CF, Tsai, HP, Chen, HS, Chang, Y, et al.. miRNA-199a-5p regulates VEGFA in endometrial mesenchymal stem cells and contributes to the pathogenesis of endometriosis. J Pathol 2014;232:330–43. https://doi.org/10.1002/path.4295.Search in Google Scholar PubMed
27. Lan, H, Zou, M, Zhu, F, Chen, H, Wang, T, Huang, X. Pro-angiogenic role of ZEB1 in skin wound healing by upregulating VEGFA via microRNA-206 suppression. Exp Dermatol 2022;31:1392–401. https://doi.org/10.1111/exd.14607.Search in Google Scholar PubMed
28. Dong, S, Wang, J, Guo, Z, Zhang, Y, Zha, W, Wang, Y, et al.. Efficient delivery of VEGFA mRNA for promoting wound healing via ionizable lipid nanoparticles. Bioorg Med Chem 2023;78:117135. https://doi.org/10.1016/j.bmc.2022.117135.Search in Google Scholar PubMed
29. Lu, YA, Jiang, Y, Yang, HW, Hwang, J, Jeon, YJ, Ryu, B. Diphlorethohydroxycarmalol isolated from Ishige okamurae exerts vasodilatory effects via calcium signaling and PI3K/Akt/eNOS pathway. Int J Mol Sci 2021;22:1610. https://doi.org/10.3390/ijms22041610.Search in Google Scholar PubMed PubMed Central
30. Qu, K, Cha, H, Ru, Y, Que, H, Xing, M. Buxuhuayu decoction accelerates angiogenesis by activating the PI3K-Akt-eNOS signalling pathway in a streptozotocin-induced diabetic ulcer rat model. J Ethnopharmacol 2021;273:113824. https://doi.org/10.1016/j.jep.2021.113824.Search in Google Scholar PubMed
31. Lin, F, Yang, Y, Wei, S, Huang, X, Peng, Z, Ke, X, et al.. Hydrogen sulfide protects against high glucose-induced human umbilical vein endothelial cell injury through activating PI3K/Akt/eNOS pathway. Drug Des Dev Ther 2020;14:621–33. https://doi.org/10.2147/dddt.s242521.Search in Google Scholar PubMed PubMed Central
32. Wang, B, Chen, S, Zhao, JQ, Xiang, BL, Gu, X, Zou, F, et al.. ADAMTS-1 inhibits angiogenesis via the PI3K/Akt-eNOS-VEGF pathway in lung cancer cells. Transl Cancer Res 2019;8:2725–35. https://doi.org/10.21037/tcr.2019.10.34.Search in Google Scholar PubMed PubMed Central
33. Hu, Y, Tao, R, Chen, L, Xiong, Y, Xue, H, Hu, L, et al.. Exosomes derived from pioglitazone-pretreated MSCs accelerate diabetic wound healing through enhancing angiogenesis. J Nanobiotechnol 2021;19:150. https://doi.org/10.1186/s12951-021-00894-5.Search in Google Scholar PubMed PubMed Central
34. Xu, X, Yang, WH, Miao, ZW, Zhang, CY, Cheng, YJ, Chen, Y, et al.. Modified Hongyu Decoction promotes wound healing by activating the VEGF/PI3K/Akt signaling pathway. Acta Biochim Pol 2023;70:843–53. https://doi.org/10.18388/abp.2020_6674.Search in Google Scholar PubMed
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