高压氧治疗通过HIF-1/NF-κB/Snail抑制瘢痕疙瘩上皮间质化的研究

Keloid represent a kind of superficial benign tumor that grows aggressively. At present, there is few non-surgical way that can cure or prevent keloid effectively. The aggressive growth of keloid is related to its epithelial-mesenchymal transition(EMT).The low-oxygen environment of keloid organization is a very important inducement of EMT.Hyperbaric oxygen (HBO) is a mature physical treatment method with little trauma, until now there is no report of the usage and mechanism in the field of using HBO to treat keloid. Through the clinical observation we found that the recurrence rate of the keloid treated by resection, radiotherapy and HBO can be reduced effectively. According to the research that HBO can improve oxygen environment of keloid tissue and affect the expression of EMT regulator and tagged molecule such as HIF-1, E-cadherin, we present a deduction that HBO can restrain EMT of keloid tissue via affecting the EMT regulator and tagged molecule. In this study, we will study the blood supply, clinical symptoms and regulators such as HIF-1, NF-κB and Snail of keloid tissue from keloid patients and nude mouse in which keloid tissue is transplanted from keloid patient. Then we will confirm HBO’s effect on EMT phenomenon via studying the expression of related molecular in keloid epithelial cells and fibroblast after hypobaric oxygen(LBO) or HBO. After all these researches , we look forward to clarify the molecular regulation mechanism which HBO therapy caused to EMT by restraining HIF-1, NF-κB and Snail in process of EMT.

瘢痕疙瘩是一种呈侵袭性生长的体表良性肿瘤，其发生发展和表皮细胞间质化（EMT）有关，而组织内低氧环境是EMT的重要诱因。高压氧是一种成熟的非手术治疗技术，其在瘢痕疙瘩治疗领域的应用及作用机制还没有研究报道。我们通过瘢痕疙瘩切除放疗后辅助高压氧治疗能有效降低瘢痕疙瘩复发率的临床观察，根据高压氧治疗能改善瘢痕疙瘩组织内氧环境、影响HIF-1、E-cadherin等EMT调控及标志分子表达的前期研究，提出了高压氧治疗能通过影响瘢痕疙瘩EMT相关调控分子抑制瘢痕疙瘩EMT的推论。本项目将通过高压氧治疗对瘢痕疙瘩病人瘢痕组织及裸鼠移植人瘢痕组织的血供及HIF-1、NF-κB、Snail等EMT调控分子影响的研究，并进一步通过瘢痕疙瘩表皮细胞、成纤维细胞在低氧和高压氧环境单独及共培养后相关分子表达的研究，确定高压氧对瘢痕疙瘩EMT现象的作用，阐明高压氧治疗抑制瘢痕疙瘩EMT现象及瘢痕增生的作用机制。

研究背景.瘢痕疙瘩是肿瘤样生长的异常纤维增生，其侵袭性生长和瘢痕疙瘩的表皮细胞间质化（EMT）现象有关，而瘢痕疙瘩的组织内环境低氧环境是EMT的重要诱因。高压氧治疗是一种成熟的非手术治疗技术，其在瘢痕疙瘩治疗领域的应用及作用机制还没有研究报道。.主要研究内容.根据高压氧治疗能改善瘢痕疙瘩组织内氧环境、影响HIF-1、E-cadherin等EMT调控及标志分子的表达，提出了高压氧治疗能通过影响瘢痕疙瘩EMT相关调控分子抑制瘢痕疙瘩EMT的推论。本项目通过高压氧治疗对瘢痕疙瘩病人瘢痕组织及裸鼠移植人瘢痕组织的血供及HIF-1、NF-κB等EMT调控分子影响的研究，并进一步通过瘢痕疙瘩表皮细胞低氧和高压氧环境培养后相关分子的表达研究，确定高压氧对瘢痕疙瘩EMT现象的作用，阐明高压氧治疗抑制瘢痕疙瘩EMT现象的分子机制。.研究结果.与瘢痕疙瘩组比较，高压氧治疗组vimentin、vibronectin、VEGF、HIF-1a表达水平较低，而E-cadherin、ZO-1表达水平明显高于K组。HBOT后E-cadherin和ZO-1的mRNA表达也增加。高压氧治疗组CD31表达明显增多（p=0.04），ET-1有增多趋势（p=0.06）。HIF-1α与Cx43、TPH1、TRPV1均正相关趋势，但无统计学意义。高压氧疗通过降低Cx43、TPH1、HIF-1α表达来缓解瘢痕疙瘩瘙痒疼痛症状，同时能上调CD31，增加血管密度。HIF-1α呈现出与Cx43、TRV1、TPH1的正相关趋势，HIF-1α可能为瘢痕疙瘩瘙痒的调控分子之一。.研究意义.瘢痕疙瘩患者在接受HBOT治疗后，与EMT现象相关的因子表达水平明显逆转，提示HBOT可能是对抗瘢痕疙瘩患者EMT现象的有效治疗方法。HIF-1α可能为瘢痕疙瘩瘙痒的调控分子之一。