HO1工程化无细胞提取液联合石墨烯远红外光促糖尿病溃疡修复的疗效及机制

温晴; 范睿诚; 杨艺; 李红丽; 熊玮

doi:10.11659/jjssx.01E023063

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HO1工程化无细胞提取液联合石墨烯远红外光促糖尿病溃疡修复的疗效及机制

基础研究 | 更新时间：2023-04-24

- HO1工程化无细胞提取液联合石墨烯远红外光促糖尿病溃疡修复的疗效及机制
- Efficacy and mechanism of HO1 engineered cell-free extract combined with graphene far-infrared radiation in promoting the repair of diabetic ulcer
- 局解手术学杂志 2023年32卷第4期页码：285-292
- 作者机构：
  
  1.陆军军医大学第一附属医院老年与特勤医学科，重庆 400038
  2.陆军军医大学基础医学院组织学与胚胎学教研室/重庆市神经科学重点实验室，重庆 400038
  3.陆军军医大学基础医学院基础医学教学实验中心，重庆 400038
- 作者简介：
  
  熊玮，E-mail：xiongwei64@126.com
- 基金信息：
  
  国家重点研发计划(2018YFC2000301)
- DOI：10.11659/jjssx.01E023063
  中图分类号： R587.2
- 纸质出版日期：2023-04-25，
  
  收稿日期：2023-01-13，
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温晴,范睿诚,杨艺等.HO1工程化无细胞提取液联合石墨烯远红外光促糖尿病溃疡修复的疗效及机制[J].局解手术学杂志,2023,32(04):285-292.

WEN Qing,FAN Rui-cheng,YANG Yi,et al.Efficacy and mechanism of HO1 engineered cell-free extract combined with graphene far-infrared radiation in promoting the repair of diabetic ulcer[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(04):285-292.
温晴,范睿诚,杨艺等.HO1工程化无细胞提取液联合石墨烯远红外光促糖尿病溃疡修复的疗效及机制[J].局解手术学杂志,2023,32(04):285-292. DOI： 10.11659/jjssx.01E023063.

WEN Qing,FAN Rui-cheng,YANG Yi,et al.Efficacy and mechanism of HO1 engineered cell-free extract combined with graphene far-infrared radiation in promoting the repair of diabetic ulcer[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(04):285-292. DOI： 10.11659/jjssx.01E023063.

摘要

目的

探讨HO1工程化无细胞提取液（CE）联合石墨烯远红外光（FIR）在糖尿病溃疡创面修复中的疗效及机制。

方法

将40只6~8周龄雄性C57BL/6小鼠建立正常创面模型（对照组，

=8）及糖尿病溃疡模型（

=32），其中将糖尿病溃疡模型小鼠随机分为糖尿病组、

HO1

BMSCs-CE组、FIR组、联合组。构建过表达血红素氧合酶1（HO1）的骨髓间充质干细胞（BMSCs），获取工程化富含HO1蛋白的无细胞提取液（

HO1

BMSCs-CE），运用Western blot、ELISA、RT-PCR、Masson染色、免疫组化等检测各组创面愈合率、组织修复情况、创面氧化应激及炎症水平等。

结果

Western blot及ELISA检测结果显示，

HO1

BMSCs-CE中HO1、表皮生长因子（EGF）、转化生长因子-β（TGF-β）表达水平显著高于BMSCs-CE（

0.05）。治疗后第5、11天，联合组的创面愈合率均显著高于

HO1

BMSCs-CE组和FIR组（

0.05）。与

HO1

BMSCs-CE组及FIR组相比，联合组在治疗后第3、7、11天创缘组织中Ca

浓度较高，TNF-α、IL-6表达水平较低，而ROS水平仅在治疗后第3、7天较低（

0.05）。治疗后第14天，联合组创面组织的表皮厚度较厚，真皮Masson染色胶原容积分数和VEGF免疫阳性信号OD值均显著高于FIR组和

HO1

BMSCs-CE组（

0.05）；治疗后第7天，联合组创面组织中凋亡相关蛋白Caspase-3表达水平显著低于FIR组和

HO1

BMSCs-CE组（

0.05）。

结论

糖尿病溃疡创面中运用工程化富含HO1蛋白的

HO1

BMSCs-CE联合石墨烯FIR治疗可明显抑制局部组织氧化应激反应，减少炎症因子释放和细胞凋亡发生，加快创面区域皮肤组织上皮化形成、真皮胶原和血管增生，较

HO1

BMSCs-CE或FIR治疗的促愈疗效更为突出，为探索临床糖尿病难愈性创面的新型联合治疗提供了基础研究证据。

Abstract

Objective

To investigate the efficacy and mechanism of HO1 engineered cell-free extract（CE） combined with graphene far-infrared radiation （FIR） in the repair of diabetic ulcer wounds.

Methods

A total of 40 male C57BL/6 mice （6 to 8 weeks old） were used to establish normal wound model （the control group，

=8） and diabetic ulcer model （

=32）. The diabetic ulcer model mice were randomly divided into the diabetes group， the

HO1

BMSCs-CE group， the FIR group and the combined group. The bone marrow mesenchymal stem cells （BMSCs） overexpressing heme oxygenase-1（HO1） was constructed to obtain engineered cell-free extract rich in HO1 protein （

HO1

BMSCs-CE）. Western blot， ELISA， RT-PCR， Masson staining and immunohistochemistry were used to detect the wound healing rate， tissue healing status， wound oxidative stress and inflammation levels in each group.

Results

The results of Western blot and ELISA showed that the expression levels of HO1， epidermal growth factor （EGF） and transforming growth factor-β （TGF-β） in

HO1

BMSCs-CE were significantly higher than those in BMSCs-CE（

0.05）. The wound healing rate 5 and 11 days after treatment in the combined group were higher than those in the

HO1

BMSCs-CE group and the FIR group（

0.05）. Compared with the

HO1

BMSCs-CE group and the FIR group， the Ca

concentration of the wound margin tissue 3， 7 and 11 days after treatment in the combined group were higher， and the expression levels of TNF-α and IL-6 were lower，while the expression levels of ROS 3 and 7 days after treatment were lower （

0.05）. The thickness of epidermis of the wound tissue 14 days after treatment in the combined group was thick， and the Masson staining collagen volume fraction in dermis and the OD value of VEGF immunopositive signal were significantly higher than those in the FIR group and the

HO1

BMSCs-CE group （

0.05）. The expression levels of apoptosis-related protein Caspase-3 of the wound tissue 7 days after treatment in the combined group was significantly lower than those in the FIR group and the

HO1

BMSCs-CE group （

0.05）.

Conclusion

The application of engineered

HO1

BMSCs-CE rich in HO1 protein combined with graphene FIR in diabetic ulcer wounds can significantly inhibit the oxidative stress response of local tissue， reduce the release of inflammatory factors and the occurrence of cell apoptosis， and accelerate the epithelization formation of skin tissue and the proliferation of dermal collagen and blood vessels in the wound area， which has more prominent curative effect than the efficacy of

HO1

BMSCs-CE or FIR therapy alone， and provide basic research evidence for the clinical exploration of new combined treatment for refractory diabetic wounds.

关键词

糖尿病溃疡骨髓间充质干细胞无细胞提取液血红素氧合酶1石墨烯远红外光

Keywords

diabetic ulcerbone marrow mesenchymal stem cellscell-free extractheme oxygenase 1graphene far-infrared radiation

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