From the Department of Plastic Surgery, Peking University Third Hospital.
Plast Reconstr Surg. 2023 Jun 1;151(6):970e-980e. doi: 10.1097/PRS.0000000000010137. Epub 2023 Jan 2.
Aerobic glycolysis (the Warburg effect) may play an important role in keloid pathogenesis, which may be aggravated by the hypoxic microenvironment in keloids. Phosphoglycerate kinase 1 (PGK1), a key glycolytic enzyme, is essential for cellular aerobic glycolysis, but its role in keloid formation remains unknown. This study aimed to detect PGK1 expression in keloid tissue and investigate the effects of inhibiting PGK1 expression on keloid fibroblasts (KFbs) under hypoxia and normoxia.
Normal skin and keloid samples were separated into two parts, one was used for immunohistochemistry, and one for primary cell culture. PGK1 tissue expression was detected by immunohistochemistry. Reverse-transcriptase polymerase chain reaction and Western blotting were used to detect PGK1, GLUT1, LDHA, and COL1 expression, and glucose uptake and lactate production were detected with a microplate reader. Cell proliferation and apoptosis were investigated with IncuCyte and flow cytometry. Cell migration and invasion were detected with Transwell assays. Glycolytic function was explored with the Seahorse XF96 system.
Immunohistochemistry showed PGK1 overexpression in keloid tissue compared with normal skin tissue ( P < 0.05). Consistently, PGK1 expression was significantly higher in KFbs than in normal skin fibroblasts (NFbs), and hypoxia stimulated PGK1 expression in KFbs and NFbs ( P < 0.05). PGK1 knockdown significantly inhibited KFb glycolysis, proliferation, migration, invasion, glucose consumption, and lactate production ( P < 0.05). Furthermore, GLUT1, LDHA, and COL1 expression was decreased in KFbs compared with NFbs ( P < 0.05). In addition, suppressing PGK1 may mediate the PI3K/AKT pathway to down-regulate GLUT1, LDHA, and COL1 expression ( P < 0.05).
These findings provide new evidence that suppressing PGK1, inhibiting glycolysis, reduces KFb proliferation, migration, invasion, and type I collagen expression. Targeting PGK1 to inhibit the Warburg effect may be a new therapeutic strategy for keloids.
This article may provide new suggestions into the pathogenesis and treatment of keloids.
CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.
有氧糖酵解（瓦博格效应）可能在瘢痕疙瘩发病机制中起重要作用，而瘢痕疙瘩中的缺氧微环境可能会加重这种作用。磷酸甘油酸激酶 1（PGK1）是一种关键的糖酵解酶，对于细胞有氧糖酵解至关重要，但它在瘢痕疙瘩形成中的作用尚不清楚。本研究旨在检测瘢痕疙瘩组织中的 PGK1 表达，并研究在缺氧和常氧条件下抑制 PGK1 表达对瘢痕疙瘩成纤维细胞（KFbs）的影响。
将正常皮肤和瘢痕疙瘩标本分为两部分，一部分用于免疫组织化学检测，一部分用于原代细胞培养。通过免疫组织化学检测 PGK1 组织表达。采用逆转录聚合酶链反应和 Western blot 检测 PGK1、GLUT1、LDHA 和 COL1 的表达，并使用微孔板读数仪检测葡萄糖摄取和乳酸生成。采用 IncuCyte 和流式细胞术研究细胞增殖和凋亡。用 Transwell 测定法检测细胞迁移和侵袭。通过 Seahorse XF96 系统探索糖酵解功能。
免疫组织化学显示，与正常皮肤组织相比，瘢痕疙瘩组织中 PGK1 过度表达（P < 0.05）。同样，KFbs 中的 PGK1 表达明显高于正常皮肤成纤维细胞（NFbs），并且缺氧刺激 KFbs 和 NFbs 中 PGK1 的表达（P < 0.05）。PGK1 敲低显著抑制 KFb 糖酵解、增殖、迁移、侵袭、葡萄糖消耗和乳酸生成（P < 0.05）。此外，KFbs 中的 GLUT1、LDHA 和 COL1 表达均低于 NFbs（P < 0.05）。此外，抑制 PGK1 可能通过介导 PI3K/AKT 通路下调 GLUT1、LDHA 和 COL1 的表达（P < 0.05）。
这些发现为抑制 PGK1、抑制糖酵解、降低 KFb 增殖、迁移、侵袭和 I 型胶原表达提供了新的证据。针对 PGK1 抑制瓦博格效应可能是瘢痕疙瘩的一种新的治疗策略。
本文可能为瘢痕疙瘩的发病机制和治疗提供新的建议。
临床问题/证据水平：治疗，V。
