牡蛎酶解产物促进小鼠皮肤创伤愈合效果研究

Effects of oyster enzymatic hydrolysate on wound healing of mouse skin

为探讨香港牡蛎Crossostrea sikamea酶解产物(enzymolysis products of oyster,EPO)对小鼠皮肤软组织开放性创伤的愈合效果,构建直径约0.8 cm的小鼠(体质量为20 g±2 g)背部全皮层创伤模型,试验设每天涂抹30~35 mg EPO(肽含量为1.5~2.5 mg)的给药组和每天涂抹2~3 mg云南白药的阳性对照组,以及不做任何处理的阴性对照组,采用体外抗菌、体表促凝血和小鼠皮肤愈合试验,进行了EPO对小鼠皮肤创伤愈合作用效果及作用阶段的研究。结果表明:EPO无促凝血作用,且对13种常见革兰氏阴性和阳性菌株无抑制作用;各试验组动物在第2天时皆已结疤,第6、10、14天时给药组创伤愈合率均显著高于阴性对照组(P<0.05);EPO可显著提高瘢痕收缩率(P<0.05);生化指标测试结果显示,EPO对炎症因子白介素-6(interleukin 6,IL-6)生成无显著性影响(P>0.05),但第3天时可显著促进白介素-10(interleukin 10,IL-10)分泌(P<0.05);给药组碱性成纤维细胞生长因子(FGF-2)含量显著低于两个对照组(P<0.05),转化生长因子β(TGF-β)含量显著高于阴性对照组(P<0.05),细胞生长周期素1(CCND1)含量显著高于两个对照组(P<0.05);给药后第7天,给药组羟脯氨酸含量较阴性对照组降低,第14天时较阴性对照组升高,但均无显著性差异(P>0.05)。研究表明,涂抹EPO可以加快小鼠软组织开放性创伤愈合,对浅表瘢痕增生具有一定的抑制作用。

Trace radial diffusion method and half dilution method were used to investigate in vitro the antibacterial effects of enzymolysis products of oyster(EPO)prepared from Hong Kong oyster Crassostrea sikamea by crushing and beating,and enzymatic hydrolysis of animal protease at a dose of 1000 U/g protein for 5 hours,and effects of the EPO on bleeding time and blood loss of mouse tail wound.In the open wound healing test,the skin soft tissue of SPF male mouse with body weight of(20±2)g cut off about 0.8 cm in diameter from the dorsal skin were smeared by 30-35 mg of EPO containing peptide of 1.5-2.5 mg(drug-administered group)daily,by 2-3 mg of Yunnan Baiyao daily(positive control)and without any smearing(negative control)to investigate the effects of EPO on the open soft tissue would healing.The results showed that EPO did not lead to promote procoagulant effect and inhibitory effect on 13 test common gram and positive bacteria.The animals in each test group had crusted on the 2nd day.On the 6th,10th and 14th day,however,significantly higher wound healing rate was observed in the drug-administered group than that in the negative control group(P<0.05).Biochemical indicators showed that EPO had no significant effect on the production of inflammatory factor interleukin 6(IL-6)(P>0.05),with promoting interleukin 10(IL-10)secretion(P<0.05)on the third day.Compared with the two control groups,there was significantly lower fibroblast growth factor 2(FGF-2)content in drug-administered group than that in the two control groups(P<0.05).The transforming growth factor-β(TGF-β)and cyclin D1(CCND1)contents were shown to be significantly higher in the drug-administered group than that in the negative control groups(P<0.05),without significant difference in the epidermal growth factor(EGF)content in the drug-administered group(P>0.05).On the 7th day after administration,there was lower hydroxyproline content in the drug-administered group than that in negative control(P>0.05),elevated hydroxyprolin content in the negative control,without significant difference between the two groups(P<0.05)on the 14th day.The findings indicate that EPO functions as acceleration of wound healing in soft tissue of mouse and has a certain inhibitory effect on superficial scar hyperplasia of a mice.