牡蛎酶解液对2型糖尿病小鼠皮肤软组织创伤愈合修复的作用及其机制研究

Effects of oyster hydrolysate on the healing of skin and soft tissue injuries in type 2 diabetic mice and its mechanisms

摘要:

目的 探究牡蛎酶解液对2型糖尿病小鼠皮肤软组织创伤愈合修复的作用，并分析相关机制。方法 将50只成功复制的2型糖尿病皮肤软组织创伤小鼠随机分为模型对照组、阳性对照组，以及牡蛎酶解液低、中、高剂量组，每组10只，其中牡蛎酶解液低、中、高剂量组小鼠于模型复制成功后次日分别给予0.25 g/10（g·d）、0.50 g/10（g·d）、1.00 g/10（g·d）牡蛎酶解液灌胃处理；模型对照组、阳性对照组同时间分别给予等量生理盐水、初元Ⅰ型产品0.1 mL/10（g·d）灌胃处理。分别于术后第3天、7天、14天观察小鼠创面并计算创面愈合率；苏木精-伊红（HE）染色观察模型复制后第14天小鼠创面组织病理变化；酶联免疫吸附试验（ELISA）检测创面组织白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）水平；Western blotting检测创面组织基质细胞衍生因子-1（SDF-1）、趋化因子受体4（CXCR4）蛋白的表达。结果 牡蛎酶解液低、中、高剂量组、阳性对照组和模型对照组小鼠术后3 d、7 d、14 d的创面愈合率比较，采用重复测量设计的方差分析，结果 ①不同时间点小鼠创面愈合率有差异（F =12.978，P =0.000）；②5组小鼠创面愈合率有差异（F =16.836，P =0.000）；③5组小鼠创面愈合率变化趋势有差异（F =22.128，P =0.000）。HE染色结果显示，与模型对照组比较，术后第3天、7天、14天，牡蛎酶解液低、中、高剂量组小鼠新生肉芽组织，毛囊、汗腺细胞和导管增生均较多。与模型对照组比较，牡蛎酶解液低、中、高剂量组小鼠创面愈合率、创面组织SDF-1、CXCR-4蛋白相对表达量升高（P <0.05），炎症因子IL-6、TNF-α水平降低（P <0.05），呈剂量依赖性。结论 牡蛎酶解液能促进2型糖尿病小鼠皮肤软组织创伤愈合修复，可能是通过激活SDF-1/CXCR-4通路进而抑制创面组织炎症反应来实现的。

Abstract:

Objective To explore the effects of oyster hydrolysate on the healing of skin and soft tissue injuries in type 2 diabetic mice and to analyze the underlying mechanisms.Methods A total of 50 type 2 diabetic mice with skin and soft tissue injuries were randomly divided into 5 groups, including model control group, positive control group and low-, medium- and high-dose oyster hydrolysate groups, with 10 mice in each group. Mice in low-, medium- and high-dose oyster hydrolysate groups were given 0.25 g/ 10 (g·d), 0.50 g/10 (g·d), and 1.00 g/ 10 (g·d) of oyster hydrolysate by gavage on the next day after establishing the models, while the mice in the model control group and the positive control group were given the same amount of normal saline and 0.1 mL / 10 (g·d) of Chueun Composite Peptide Nutritional Diet I by gavage on the same day. The wounds on mice were observed and the wound area was calculated on the 3rd, 7th and 14th day after administration. Hematoxylin and eosin (HE) staining was used to observe the histopathological changes of the wound tissues on the 14th day after establishing the models. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The protein expressions of stromal cell-derived factor-1 (SDF-1) and chemokine receptor 4 (CXCR4) in wound tissues were detected by Western blotting.Results The rate of wound healing in all the groups on the 3rd, 7th and 14th day after establishing the models were compared via repeated measures analysis of variance, and the results exhibited that the rate of wound healing was different among the time points (F = 12.978, P = 0.000) and among the groups (F = 16.836, P = 0.000), and that the change trend of the rate of wound healing was also different among the groups (F = 22.128, P = 0.000). The HE staining showed that the frequency of newly formed granulation tissues, hair follicles, sweat gland cells and ductal proliferation on the 3rd, 7th and 14th day after establishing the models was higher in the low-, medium- and high-dose oyster hydrolysate groups relative to that in the model control group. Compared with the model control group, the wound healing rate and the protein expressions of SDF-1 and CXCR-4 were higher, yet the levels of IL-6 and TNF-α were lower in the low-, medium- and high-dose oyster hydrolysate groups in a dose-dependent manner (P < 0.05).Conclusions The oyster hydrolysate can accelerate the healing of skin and soft tissue injuries in type 2 diabetic mice, which may be achieved by activating the SDF-1/CXCR-4 pathway and inhibiting inflammatory responses in wound tissues.