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血清蛋白质组学分析在链脲佐菌素诱导的糖尿病模型中体外冲击波治疗增强糖尿病创面愈合的作用。

Serum proteomic analysis of extracorporeal shock wave therapy-enhanced diabetic wound healing in a streptozotocin-induced diabetes model.

机构信息

Tao-Yuan and Kaohsiung, Taiwan From the Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University; and the Department of Plastic and Reconstructive Surgery, Medical Research, and the Department of Orthopedics, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine.

出版信息

Plast Reconstr Surg. 2014 Jan;133(1):59-68. doi: 10.1097/01.prs.0000439050.08733.cf.

Abstract

BACKGROUND

Previous studies have demonstrated that extracorporeal shock wave therapy has a significant positive effect on accelerating diabetic wound healing. However, the systemic effect after therapy is still unclear.

METHODS

This study investigated the plasma protein expression in the extracorporeal shock wave therapy group and diabetic controls using proteomic study. A dorsal skin defect (6 × 5 cm) in a streptozotocin-induced diabetic Wistar rat model was used. Diabetic rats receiving either no therapy or extracorporeal shock wave therapy after wounding were analyzed. The spots of interest were subjected to in-gel trypsin digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry to elucidate the peptide mass fingerprints. The mass spectrometric characteristics of the identified proteins, including their theoretical isoelectric points, molecular weights, sequence coverage, and Mascot score, were analyzed. Protein expression was validated using immunohistochemical analysis of topical periwounding tissues.

RESULTS

The proteomic study revealed that at days 3 and 10 after therapy rats had significantly higher abundance of haptoglobin and significantly lower levels of the vitamin D-binding protein precursor as compared with the diabetic controls. Immunohistochemical staining of topical periwounding tissue also revealed significant upregulation of haptoglobin and downregulation of vitamin D-binding protein expression in the extracorporeal shock wave therapy group, which was consistent with the systemic proteome study.

CONCLUSION

Proteome analyses demonstrated an upregulation of haptoglobin and a downregulation of vitamin D-binding protein in extracorporeal shock wave therapy-enhanced diabetic wound healing.

摘要

背景

先前的研究表明,体外冲击波治疗对加速糖尿病创面愈合有显著的积极影响。然而,治疗后的全身效应仍不清楚。

方法

本研究采用蛋白质组学研究方法,研究了体外冲击波治疗组和糖尿病对照组的血浆蛋白表达。在链脲佐菌素诱导的糖尿病 Wistar 大鼠模型中建立背部皮肤缺损(6×5cm)。对创伤后未接受治疗或接受体外冲击波治疗的糖尿病大鼠进行分析。对感兴趣的斑点进行胶内胰蛋白酶消化和基质辅助激光解吸电离飞行时间质谱分析,以阐明肽质量指纹图谱。对鉴定蛋白的质谱特性,包括其理论等电点、分子量、序列覆盖率和 Mascot 评分进行分析。通过对创面周围组织的免疫组织化学分析验证蛋白表达。

结果

蛋白质组学研究表明,与糖尿病对照组相比,治疗后第 3 天和第 10 天,大鼠的触珠蛋白含量显著升高,维生素 D 结合蛋白前体水平显著降低。对创面周围组织的免疫组织化学染色也显示,体外冲击波治疗组触珠蛋白表达显著上调,维生素 D 结合蛋白表达下调,与系统蛋白质组学研究结果一致。

结论

蛋白质组学分析表明,体外冲击波治疗增强糖尿病创面愈合过程中触珠蛋白上调,维生素 D 结合蛋白下调。

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