珍珠贝外套膜酶解产物促进小鼠皮肤创伤愈合作用研究

Effects of enzymatic hydrolysates from mantle of pearl oyster Pinctada martensii on wound healing of mouse skin

为探讨珍珠贝Pinctada martensii外套膜酶解产物(enzymatic hydrolysis from mantle of pearl oyster,EHM)对小鼠皮肤创伤的促愈合效果,采用小鼠(体质量20 g±2 g)背部全皮层创伤模型,连续灌胃给药14 d,隔天观察伤口愈合情况,计算创伤愈合率和瘢痕缩小率,并于造模后的第3、7和14天分批处死动物,测定伤口边缘皮肤组织中白介素-6(interleukin 6,IL-6)、白介素-10(interleukin 10,IL-10)、转化生长因子β(transforming growth factor-β,TGF-β)、碱性成纤维细胞生长因子(fibroblast growth factor 2,FGF-2)、表皮细胞生长因子(epidermal growth factor,EGF)、细胞生长周期素1(cyclin D1,CCND1)和羟脯氨酸含量,对EHM的作用效果和作用阶段进行研究。结果表明:各试验组动物在第2天时皆已结疤,第6、10天时,各EHM给药组创伤愈合率均高于阴性对照组,第14天时,低剂量组的愈合率达100%;EHM可提高瘢痕收缩率,其对炎症因子IL-6生成无显著性影响(P>0.05),但能极显著促进IL-10分泌(P<0.01);与阴性对照组相比,高剂量EHM给药组的TGF-β、CCND1和EGF含量均显著增加(P<0.05),而FGF-2含量则无显著性差异(P>0.05);给药后第14天,各EHM给药组均能显著提高皮肤组织中的羟脯氨酸含量(P<0.05)。研究表明,EHM具有抑制炎症作用并能促进生长因子分泌及胶原蛋白的生成,从而加快小鼠软组织开放性创伤愈合,EHM还对浅表瘢痕增生具有一定的抑制作用。

SPF male mice were randomly divided into negative control group, positive control group and experimental groups(low, medium and high), and the mice with incision dorsal wound model were intragastrically administrated by enzymatic hydrolysis from mantle of pearl oyster Pinctada martensii(EHM) continuously for 14 days. The wound healing rate and scar reduction rate of soft tissue were measured every other day and biochemical factors including interleukin 6(IL-6), interleukin 10(IL-10), transforming growth factor-β(TGF-β), fibroblast growth factor 2(FGF-2), epidermal growth factor(EGF), cyclin D1(CCND1) and hydroxyproline were investigated in marginal tissue of wound of the mice at the 3 rd day, 7 th day, and 14 th day to evaluate the effects of EHM on wound healing of skin in mouse. Results showed that the EHM led to improve the healing rate and scar shrinkage of wound in mouse compared with the negative control group at 6 th and 10 th day(P<0.05). The EHM was shown to effectively increase the protein expression of anti-inflammatory cytokine IL-10(P<0.01) in wound tissues, without significant effect on inflammatory factor IL-6(P>0.05). There were significantly higher contents of TGF-β, EGF and CCND1 in injured skin tissue in the high dose group than those in control group(P<0.05), without significant differencein content of FGF-2 between the test groups(P>0.05). On the 14 th day, hydroxyproline content in the skin tissue was significantly increased in each EHM groups(P<0.05). The findings indicated that EHM could inhabit inflammation, promote growth factors and collagen production, thus accelerate open wound healing of soft tissue in mice, and that EHM has a certain inhibitory effect on superficial scar hyperplasia.