方格星虫水提物促小鼠皮肤创伤愈合及作用机制的研究

Wound-healing acceleration of mice skin by Sipunculus nudus extract and its mechanism

为探讨方格星虫水提物(SNE)促进小鼠皮肤创伤愈合的活性及其机制,测定SNE的止血作用,建立小鼠全皮层创伤模型,观察伤口愈合形态并测定愈合率;采用苏木素-伊红(HE)、马森(Masson)染色、透射电镜(TEM)观察创面的组织结构;实时荧光定量聚合酶链式反应(qRT-PCR)检测细胞因子及其相关蛋白的表达水平。结果显示,SNE具有止血作用,能明显提高小鼠创伤愈合率并缩短掉痂时间(P <0.05)。研究中,SNE组小鼠表皮生长完全、创面毛细血管和胶原纤维增生明显优于空白(NT)组;第7 d,SNE明显降低白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和转化生长因子-β1(TGF-β1)的表达水平(P <0.05);第28 d,相比于NT组,SNE组的Smad7表达水平上调,TGF-βII型受体(TGF-βRII)、Ⅰ型胶原蛋白(COL1A1)和α-平滑肌肌动蛋白(α-SMA)的表达水平下调,差异具有统计学意义(P <0.05),而与云南白药组(PC组)的差异无统计学意义(P> 0.05)。结果表明,SNE具有促进伤口愈合活性和抑制增生性瘢痕生成作用,其作用机制与止血、调节炎症因子、促进胶原纤维重塑以及调控TGF-β/Smads信号通路传导有密切关联。SNE作为皮肤创伤修复和抑制瘢痕制剂,具有潜在的临床应用价值。

In order to explore the effect of Sipunculus nudus extract(SNE) on skin wound healing in mice and its mechanism, hemostasis effect of SNE was measured, the mouse skin wound model was established by full-thickness excision. The morphological changes of the wound were observed after the treatment with SNE and the healing rate was measured. The changes of wound histology were observed by hematoxylin eosin(HE) staining, Masson staining and transmission electron microscope(TEM). The expression of cell factors and related proteins was detected by quantitative real-time polymerase chain reaction(qRT-PCR). Results showed that the SNE possessed hemostatic function. SNE could obviously improve the healing rate of wound in mouse and shorten time of scab removal compared with the nonetreatment(NT) group(P < 0.05).The pathological histology analysis results showed complete epidermal regeneration, with remarkable capillary and collagen fiber observed in the SNE group. The expression level of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and transforming growth factor-β1(TGF-β1) in SNE group was significantly lower than that of the NT group on 7 d(P < 0.05). Moreover, compared with the NT group, the gene expressions level of Smad7 was significantly increased and the level of type Ⅱ TGF-β receptors(TGF-βRⅡ), collagen I(COL1 A1) and α-smooth muscle actin(α-SMA) were significantly reduced in the SNE group on 28 d(P < 0.05), but the difference was not statistically significant compared to Yunnanbaiyao group(PC group)(P > 0.05). These results indicated that SNE possessed obvious activity of accelerating wound healing and inhibiting scar formation, and its mechanism was closely related to hemostatic function, regulation of inflammatory factors, collagen deposition, collagen fiber remodeling and intervening TGF-β/Smads signal pathway. Therefore, SNE may have promising clinical applications in skin wound repair and scar inhibition.