Modulating hypertrophic scar formation by targeting endothelial transient receptor potential vanilloid-1/nuclear factor kappa-B/interleukin-6 axis to regulate angiogenesis

Burns & Trauma, vol. 14, pp. tkag009

Abstract

Abstract Background Noxious lifestyle factors including spicy diets and hot baths may lead to scar formation and recurrence. These phenomena are related to the activation of the transient receptor potential vanilloid-1 (TRPV1) cation channel. Our previous study revealed significant upregulation of TRPV1 expression in the dermis of hypertrophic scar (HS), while the exact underlying mechanism of TRPV1 activation in HS remains ill-defined. This study aims to clarify the contribution of TRPV1 activation to HS pathogenesis, particularly in relation to aberrant angiogenesis. Methods First, this study employs single-cell RNA sequencing technology to analyze the association between vascular endothelial cells and the development of HS. Complementarily, bioinformatics analysis combined with histological validation is utilized to investigate the relationship between TRPV1 channels and aberrant angiogenesis within HS formation. Furthermore, the correlation between TRPV1 activation and HS phenotypes is rigorously validated at the in vivo level. In parallel, in vitro experiments are conducted to elucidate the impact of TRPV1 channel activation on the biological behaviors and functions of vascular endothelial cells. Subsequently, key downstream signaling pathways of TRPV1 are screened, and their molecular mechanisms in regulating vascular endothelial cell-mediated angiogenesis are systematically verified. Finally, a comprehensive analysis is performed to establish the clinical relevance of the TRPV1/nuclear factor kappa-B (NF-κB)/interleukin-6 (IL-6) axis with vascularization severity and adverse prognostic outcomes in hypertrophic scarring. Results Single-cell RNA sequencing revealed significant cellular heterogeneity in vascular endothelial cells between normal skin and HS, indicating activated angiogenesis and substantial vascular endothelial cell alterations during HS development. Bulk RNA-seq and clinical analyses further confirmed this angiogenesis activation, demonstrating a close association with TRPV1 channel activation. In vivo studies established that capsaicin (CAP)-induced TRPV1 activation exacerbated HS progression through enhanced angiogenesis, whereas TRPV1 ablation or local inhibition markedly attenuated this effect. In vitro experiments demonstrated that TRPV1 activation regulated angiogenesis by promoting pro-angiogenic phenotypes. Transcriptomic analysis and functional validation identified the IL-6/signal transducer and activator of transcription 3 pathway as a downstream NF-κB-dependent pro-angiogenic axis mediated by TRPV1 in HS vascular endothelial cells. Critically, dermal overexpression of the TRPV1/NF-κB/IL-6 axis in HS patients correlated strongly with both disease severity and recurrence. Conclusions Here, we show that the development of HS is strongly correlated with endothelial angiogenic activity. TRPV1 activation by CAP enhances proangiogenic processes including endothelial proliferation, migration, and tubule formation, while reducing apoptosis through the TRPV1/NF-κB/IL-6 axis. In a rabbit ear HS model, stimulation of TRPV1 contributes to the formation of HS via the TRPV1/NF-κB/IL-6 axis, whereas pharmacological ablation of TRPV1 significantly reversed these phenotypes. These findings shed light on the underlying molecular mechanisms and provide a potential therapeutic target for HS.

Authors 10

1. Hao Ma

   Shanghai Jiao Tong University · Shanghai Ninth People's Hospital

   Affiliation as printed

   Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,

   Shanghai Institute for Plastic and Reconstructive Surgery , Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, No. 639 Zhizaoju Road, Shanghai, 200011 ,

2. Liuhanghang Cheng Aachen Department of Oral & Maxillofacial Surgery (Chair: Prof. Dr. JPR van Merkesteyn)

   Leiden University · Shanghai Jiao Tong University · Leiden University Medical Center · Shanghai Ninth People's Hospital

   Affiliation as printed

   Department of Oral and Maxillofacial Surgery, Leiden University Medical Centre , Albinusdreef 2, 2333 ZA Leiden ,

   Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,

   Shanghai Institute for Plastic and Reconstructive Surgery , Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, No. 639 Zhizaoju Road, Shanghai, 200011 ,

3. Ruoyu Ling
   Affiliation as printed

   Department of Burn and Plastic Surgery, Southern Theater General Hospital , No. 111 Liuhua Road, Guangzhou, 510010 ,

4. J W Chen

   Shanghai Jiao Tong University · Shanghai Ninth People's Hospital

   Affiliation as printed

   Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,

   Shanghai Institute for Plastic and Reconstructive Surgery , Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine, No. 639 Zhizaoju Road, Shanghai, 200011 ,

5. Shunuo Zhang

   Shanghai Jiao Tong University · Shanghai Ninth People's Hospital

   Affiliation as printed

   Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,

6. Shujing Lin

   Shanghai Jiao Tong University

   Affiliation as printed

   School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University , 800 Dongchuan Road, Shanghai, 200240 ,

7. Liang Ding

   Guangxi Normal University · State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources

   Affiliation as printed

   State Key Laboratory for Chemistry and Molecular Engineering of Medical Resources, School of Chemistry and Pharmaceutical Sciences, Guangxi Normal University , 15 Yucai Road, Guilin, 541004 ,

8. Chengliang Deng corresponding

   Zunyi Medical University · Affiliated Hospital of Zunyi Medical College

   Affiliation as printed

   Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University , 149 Dalian Road, Zunyi, 563003 ,

9. Yixin Zhang corresponding

   Shanghai Jiao Tong University · Shanghai Ninth People's Hospital

   Affiliation as printed

   Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,

10. Peiru Min corresponding

    Shanghai Jiao Tong University · Shanghai Ninth People's Hospital

    Affiliation as printed

    Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of Medicine , No. 639 Zhizaoju Road, Shanghai, 200011 ,