中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (26): 4126-4131.doi: 10.12307/2023.479

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

石香膏干预大鼠感染难愈创面的愈合机制

刘志伦1，关智宇2，蒋太平1，李成蹊1，刘昭明1   

  1. 1贵州中医药大学，贵州省贵阳市  550000；2贵州中医药大学第一附属医院骨科，贵州省贵阳市  550001
  • 收稿日期:2022-05-10 接受日期:2022-07-18 出版日期:2023-09-18 发布日期:2023-01-28
  • 通讯作者: 关智宇，博士，硕士生导师，主任医师，贵州中医药大学第一附属医院骨科，贵州省贵阳市 550001
  • 作者简介:刘志伦，男，1995年生，贵州省大方县人，彝族，在读硕士，主要从事四肢骨与关节及其相关疾病的临床和研究。
  • 基金资助:
    国家自然科学基金项目(81960813)，项目负责人：关智宇

Mechanism of stone balm on the healing of infected refractory wounds in rats

Liu Zhilun1, Guan Zhiyu2, Jiang Taiping1, Li Chengxi1, Liu Zhaoming1   

  1. 1Guizhou University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China; 2Department of Orthopedics, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China
  • Received:2022-05-10 Accepted:2022-07-18 Online:2023-09-18 Published:2023-01-28
  • Contact: Guan Zhiyu, MD, Master’s supervisor, Chief physician, Department of Orthopedics, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China
  • About author:Liu Zhilun, Master candidate, Guizhou University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81960813 (to GZY)

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摘要：

文题释义：

氧化应激损伤：是指机体在受到感染等有害刺激时，体内高活性分子如活性氧自由基和活性氮自由基产生过多，超过机体清除能力，促使机体氧化还原系统失衡，导致细胞内蛋白质、核酸的氧化性损伤，从而引起细胞凋亡和组织器官损伤。
Nrf2-ARE信号通路：是细胞抗氧化应激的关键通路，在机体处于氧化应激状态下时能够激活该通路，调控下游基因表达抗氧化酶系和Ⅱ相解毒酶来清除活性氧自由基等有害物质，减轻细胞损伤。

背景：在临床工作中，西医对于感染性皮肤难愈合创口的治疗手段较少，中医药对皮肤创面的治疗则越来越受到关注。
目的：探讨石香膏促进大鼠感染难愈性皮肤创面模型愈合的作用及机制。
方法：选取15只雄性SD大鼠，其中12只大鼠采取创面接种细菌法建立感染难愈合皮肤创面模型，按体质量随机分为模型组(不进行治疗)、贝复剂(重组牛碱性成纤维细胞成长因子)组及石香膏低剂量、高剂量组，每48 h给药一次，每组3只；剩余3只大鼠仅做创面处理，不接种细菌，作为空白对照组。用药14 d后，观察创面愈合情况，镜下观察创面组织病理变化，采用ELISA法检测创面组织超氧化物歧化酶、丙二醛因子水平，Western Blot法检测创面组织Nrf2、血红素氧合酶1蛋白表达。

结果与结论：①用药第14天时，各组创面均呈现愈合状态；与空白对照组比较，模型组创面面积稍小，但仍能看见红肿；与模型组比较，贝复剂组、石香膏高剂量组创面面积均减小，创面红肿程度减轻，石香膏低剂量组创面面积变化不大，但创面红肿程度减轻；与贝复剂组比较，石香膏高剂量组创面面积明显减小，创面基本愈合；②苏木精-伊红染色显示，与空白对照组及模型组比较，贝复剂组及石香膏低、高剂量组皮肤表层坏死物、炎性物均减少，可见大量胶原纤维、肉芽组织增生；与贝复剂组比较，石香膏高剂量组胶原纤维及肉芽组织较为密集；③与模型组比较，贝复剂组及石香膏高、低剂量组丙二醛含量下降(P < 0.05)，超氧化物歧化酶水平升高(P < 0.05)；④与模型组比较，贝复剂组Nrf2、血红素氧合酶1蛋白表达升高(P < 0.05)，石香膏高、低剂量组血红素氧合酶1 蛋白表达升高(P < 0.05)；⑤结果表明，调节Nrf2-ARE信号通路诱导ARE抗氧化反应元件产生血红素氧合酶1蛋白及超氧化物歧化酶因子，同时抑制丙二醛产生，减轻细胞氧化应激损伤，是石香膏促进创面愈合的可能机制之一。

https://orcid.org/0000-0002-8868-486X(刘志伦)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 感染, 皮肤创面, 难愈创面, 氧化应激损伤, Nrf2-ARE信号通路, 中药治疗

Abstract: BACKGROUND: In clinical work, western medicine has less treatment methods for skin-infected refractory wounds, while the treatment of skin wounds with traditional Chinese medicine has attracted more and more attention.
OBJECTIVE: To investigate the effect and mechanism of stone balm on promoting the healing of skin-infected refractory wounds in rats. 
METHODS: A total of 15 male Sprague-Dawley rats were selected. Of these rats, 12 rats were used to establish animal models of skin-infected refractory wounds and then randomly divided into model group (no intervention), recombinant bovine basic fibroblast growth factor (rb-bFGF) group, low- and high-dose stone balm groups, with 3 rats in each group. Administration was performed every 48 hours. The remaining three rats were not given inoculation of bacteria, serving as blank control group. After 14 days of medication, wound healing was observed and pathological changes in wound tissue were observed under microscope. The levels of superoxide dismutase and malondialdehyde in wound tissue were detected by ELISA, and the protein expressions of Nrf2 and heme oxygenase 1 in wound tissue were detected by western blot method.
RESULTS AND CONCLUSION: After 14 days of intervention, the wounds in each groups showed a healing state. Compared with the blank control group, the wound area was slightly smaller in the model group, but redness and swelling could still be seen. Compared with the model group, the wound area of were reduced in the rb-bFGF and high-dose stone balm groups, and wound redness and swelling was alleviated, while the wound area did not change significantly in the low-dose stone balm group but wound redness and swelling was also alleviated. Compared with the rb-bFGF group, the wound surface was significantly reduced in the high-dose Stone balm group, and the wound basically healed. Hematoxylin-eosin staining showed that compared with the blank control and model groups, the number of sphacelus and inflammatory substances on the wound surface was significantly reduced in the rb-bFGF, low- and high-dose Stone balm groups, and a large number of collagen fibers and granulation tissue hyperplasia were observed. Compared with the rb-bFGF group, collagen fibers and granulation tissue were denser in high-dose Stone balm group. Compared with the model group, the levels of malondialdehyde were significantly lower in the rb-bFGF, low- and high-dose stone balm groups (P < 0.05), while the levels of superoxide dismutase were significantly higher (P < 0.05). The protein expressions of Nrf2 and heme oxygenase 1 were significantly higher in the rb-bFGF group than the model group (P < 0.05), and the protein expression of heme oxygenase 1 was significantly higher in the high- and low-dose stone balm groups than the model group (P < 0.05). To conclude, regulation of the Nrf2-ARE signaling pathway induces the ARE antioxidant response element to produce heme oxygenase 1 protein and superoxide dismutase factor, while inhibiting the production of malondialdehyde and reducing cellular oxidative stress injury, which may be one of the possible mechanisms by which stone balm promotes wound healing.

Key words: infection, skin wound, refractory wound, oxidative stress injury, Nrf2-ARE signaling pathway, traditional Chinese medicine treatment

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