Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China.
Department of Esthetic Plastic Surgery, The First Affiliated Hospital of Nanjing University of TCM, Nanjing, Jiangsu 210029, P.R. China.
Int J Mol Med. 2018 Dec;42(6):3073-3082. doi: 10.3892/ijmm.2018.3915. Epub 2018 Oct 2.
Cutaneous ischemia‑reperfusion (I/R) injury is one of the most crucial problems in flap surgery, which affects the survival of the skin flap and patient prognosis, luteolin, a plant derived flavonoid, has previously been shown to exert a variety of beneficial effects for reducing I/R injury in several organs. The aim of the present study was to evaluate the anti‑inflammatory and anti‑oxidative stress effects of luteolin on cutaneous I/R injury. The in vitro study were performed using a permanent human immortalized epidermal keratinocyte cell line (HaCaT), cells were cultured in the presence of luteolin and were then treated with hydrogen peroxide, the cell viability, mitochondrial membrane potential and the cell survival/apoptosis related signaling pathway activation were assessed to investigate the cytoprotective effects of luteolin. For in vivo experiments, skin flap I/R injury animal model was established in Sprague‑Dawley rats, by measuring the area of flap survival, analyzing the expression of pro‑inflammatory cytokine and evaluation of the histological changes in the skin tissue, the protective effects of luteolin on skin I/R injury were investigated. The function of protein kinase B (AKT) and heme oxygenase‑1 (HO‑1) activation on luteolin mediated I/R injury protection was assessed by administration of phosphoinositide‑3‑kinase/AKT inhibitor LY294002 and HO‑1 inhibitor ZNPP. The results showed that luteolin treatment significantly increased the viability of HaCaT cells upon exposure to hydrogen peroxide, and the administration of luteolin in vivo significantly improved skin flap survival in the I/R injury rat model. The mechanisms underlying these beneficial effects included increased phosphoinositide‑3‑kinase/protein kinase B activation, improved expression of antioxidant enzyme, and scavenging the cytotoxic effects of reactive oxygen species (ROS). Taken together, the results suggested that luteolin preconditioning yielded significant protection against cutaneous I/R injury by protecting skin keratinocytes from ROS‑induced damage.
皮肤缺血再灌注（I/R）损伤是皮瓣手术中最关键的问题之一，它影响皮瓣的存活和患者的预后。木犀草素是一种植物衍生的类黄酮，先前已被证明在几种器官的 I/R 损伤中具有多种有益作用。本研究旨在评估木犀草素对皮肤 I/R 损伤的抗炎和抗氧化应激作用。体外研究使用人永生化表皮角质形成细胞系（HaCaT）进行，细胞在木犀草素存在的情况下培养，然后用过氧化氢处理，评估细胞活力、线粒体膜电位和细胞存活/凋亡相关信号通路的激活，以研究木犀草素的细胞保护作用。对于体内实验，通过测量皮瓣存活面积、分析促炎细胞因子的表达以及评估皮肤组织的组织学变化，在 Sprague-Dawley 大鼠中建立皮肤皮瓣 I/R 损伤动物模型，研究木犀草素对皮肤 I/R 损伤的保护作用。通过给予磷酸肌醇 3-激酶/蛋白激酶 B（AKT）抑制剂 LY294002 和血红素加氧酶-1（HO-1）抑制剂 ZNPP，评估 AKT 和 HO-1 激活对木犀草素介导的 I/R 损伤保护的作用。结果表明，木犀草素处理可显著增加 HaCaT 细胞在暴露于过氧化氢时的活力，体内给予木犀草素可显著改善 I/R 损伤大鼠模型中的皮瓣存活。这些有益作用的机制包括增加磷酸肌醇 3-激酶/AKT 激活、改善抗氧化酶的表达以及清除活性氧（ROS）的细胞毒性作用。总之，这些结果表明，木犀草素预处理通过保护皮肤角质形成细胞免受 ROS 诱导的损伤，对皮肤 I/R 损伤产生显著的保护作用。
