Electrical stimulation promotes wound healing and angiogenesis in diabetic rats

doi:10.3969/j.issn.2095-4344.2734

Abstract

Abstract:

BACKGROUND: Diabetic wound healing disorder is one of the common complications of diabetes, but its pathogenesis has not yet been elucidated. Electrical stimulation therapy is one of the commonly used methods in the clinical treatment of wound injury, which can effectively promote the healing of injured skin.

OBJECTIVE: To investigate the effects of electrical stimulation therapy on wound healing and angiogenesis in diabetic rats and its potential mechanism.

METHODS: Forty-two SPF male Sprague-Dawley rats were enrolled in this study. A rat model of diabetic refractory wounds was established in 24 rats by tail vein injection of streptozotocin (50 mg/kg) combined with back skin wounds, and model rats were randomly divided into model group and electrical stimulation group. Meanwhile, normal rats were taken to cause back wounds as a blank control group. The electrical stimulation group was treated with electrical stimulation for 21 days. The blank control group and the model group were fed normally. The wound healing was evaluated on the 3^rd, 7^th, 14^th and 21^stdays after treatment. After treatment, the rat serum and wound tissue were taken for index detection. The pathological morphology of rat wounds was observed by hematoxylin-eosin staining. The serum endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels were detected by enzyme-linked immunosorbent assay. The expression levels of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 proteins were detected by immunohistochemistry and western blot. An ethical approval was obtained from the Animal Experimental Ethics Committee of the Affiliated Hospital of Southwest Medical University.

RESULTS AND CONCLUSION: The wound healing rate of diabetic rats was close to 90% on the 14^th day after electrical stimulation, while the healing rate in the model group was less than 60% (P < 0.01). The serum endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels were significantly higher in the electrical stimulation group than the model group (P < 0.05 or P < 0.01). Meanwhile, compared with the model group, the electrical stimulation group had more neovascularization, larger vascular lumen, higher expression of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 around the blood vessels (P < 0.05 or P < 0.01). These findings indicate that electrical stimulation therapy can significantly promote wound healing and neovascularization in diabetic rats, and its mechanism is related to the increase of endothelial nitric oxide synthase, angiopoietin 1 and vascular endothelial growth factor levels and up-regulation of angiopoietin receptor 2 and vascular endothelial growth factor receptor 2 levels.

Key words: electrical stimulation therapy, diabetes, wound healing, angiogenesis, angiogenin, vascular endothelial growth factor

CLC Number:

Geng Kang, Ding Xiaobin, Tian Xinli, Wang Xue, Yang Yuting, Yan Hong.

Electrical stimulation promotes wound healing and angiogenesis in diabetic rats [J]. Chinese Journal of Tissue Engineering Research, 2020, 24(26): 4152-4156.

Figures/Tables 7

References

[1] International Diabetes Federation. IDF Diabetes Atlas, 8th ed. Brussels: International Diabetes Federation.2017.

[2] LIU H, DUAN Z, TANG J, et al. A short peptide from frog skin accelerates diabetic wound healing. FEBS J.2014;281(20):4633-4643.

[3] 赵敏超,赵敏越,袁伟健,等.局部应用胰岛素促进糖尿病创面愈合的研究进展[J].医学综述,2018,24(20):4078-4085.

[4] 向鹏君,季晖,顾铭.糖尿病创面的炎症机制研究进展[J].药学研究,2017, 36(11):667-670.

[5] LI W, KANDHARE AD, MUKHERJEE AA. Hesperidin, a plant flavonoid accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats: Role of TGF-ß/Smads and Ang-1/Tie-2 signaling pathways. EXCLI J.2018;17(4):399-419.

[6] RUFAIHAH AJ, JOHARI NA, VAIBAVI SR, et al. Dual delivery of VEGF and ANG-1 in ischemic hearts using an injectable hydrogel. Acta Biomater.2017;48(15):58-67.

[7] KADO M, TANAKA R, ARITA K, et al. Human peripheral blood mononuclear cells enriched in endothelial progenitor cells via quality and quantity controlled culture accelerate vascularization and wound healing in a porcine wound model.Cell Transplant.2018;27(7):1068-1079.

[8] 李少源,翟煦,荣培晶,等.电针耳甲区对2型糖尿病大鼠痛觉障碍及抑郁症状的影响[J].中医杂志,2014,55(2):148-152.

[9] KIM TH, CHO HY, LEE SM. High-voltage pulsed current stimulation enhances wound healing in diabetic rats by restoring the expression of collagen, α-smooth muscle actin, and TGF-β1. Tohoku J Exp Med. 2014;234(1):1-6.

[10] 王芳,栗勇,苏映军,等.糖尿病难愈创面应用小剂量胰岛素:上皮化、纤维化及血管新生[J].中国组织工程研究,2013,17(20):3650-3657.

[11] THAKRAL G, LAFONTAINE J, NAJAFI B, et al. Electrical stimulation to accelerate wound healing. Diabet Foot Ankle. 2013;16:4-11.

[12] VATANKHAH N, JAHANGIRI Y, LANDRY GJ, et al. Effect of systemic insulin treatment on diabetic wound healing. Wound Repair Regen. 2017;25(2):288-291.

[13] JI XY, CHEN Y, YE GH, et al. Detection of RAGE expression and its application to diabetic wound age estimation. Int J Legal Med.2017; 131(3):691-698.

[14] YE J, KANG Y, SUN X, et al. MicroRNA-155 Inhibition Promoted Wound Healing in Diabetic Rats[J]. Int J Low Extrem Wounds.2017; 16(2):74-84.

[15] ZENG Z, HUANG WD, GAO Q, et al. Arnebin-1 promotes angiogenesis by inducing eNOS, VEGF and HIF-1α expression through the PI3K-dependent pathway. Int J Mol Med.2015;36(3):685-697.

[16] 吴子涵,李高峰.瘢痕成熟过程中血管生成素1表达与瘢痕血管的变化[J].中国组织工程研究,2017,32(21):5158-5163.

[17] BOUCHERAT O, FRANCO-MONTOYA ML, DELACOURT C, et al. Defective angiogenesis in hypoplastic human fetal lungs correlates with nitric oxide synthase deficiency that occurs despite enhanced angiopoietin-2 and VEGF. Am J Physiol Lung Cell Mol Physiol.2010; 298(6): 849-856.

[18] LI J, ZHANG Y, LI C, et al. HSPA12B attenuates cardiac dysfunction and remodelling after myocardial infarction through an eNOS-dependent mechanism. Cardiovasc Res.2013;99(4):674-684.

[19] SADATI SM, RADFAR M, HAMIDI AK, et al. Association Between the Polymorphism of Glu298Asp in Exon 7 of the eNOS Gene With Foot Ulcer and Oxidative Stress in Adult Patients With Type 2 Diabetes. Can J Diabetes.2018; 42(1): 18-22.

[20] WANG X, XIU P, WANG F, et al. P18 peptide, a functional fragment of pigment epithelial-derived factor, inhibits angiogenesis in hepatocellular carcinoma via modulating VEGF/VEGFR2 signalling pathway. Oncol Rep.2017;38(2):755-766.

[21] MENG Q, SHAO L, LUO X, et al. Expressions of VEGF-A and VEGFR-2 in placentae from GDM pregnancies.Reprod Biol Endocrinol. 2016;14(1):61-70.

[22] BITTO A, MINUTOLI L, GALEANO MR, et al. Angiopoietin-1 gene transfer improves impaired wound healing in genetically diabetic mice without increasing VEGF expression. Clin Sci.2008;114(12): 707-718.

[23] CHEN JX, TUO Q, LIAO DF. Inhibition of protein tyrosine phosphatase improves angiogenesis via enhancing Ang-1/Tie-2 signaling in diabetes. Exp Diabetes Res.2012;2012:836759.

[24] CHAN W, ISMAIL H, MAYAKI D, et al. Fibulin-5 Regulates Angiopoietin-1/ Tie-2 Receptor Signaling in Endothelial Cells. PLoS One.2016;11(6): e0156994.

[25] 刘帅,刘学政.Ang-1/Tie2系统与病理性血管形成的关系[J].解剖科学进展, 2010,4(1):85-88.

[26] 胡夏晓,张文倩,刘炳男. Ang-1和Tie-2在血管生成中的作用及中医药干预研究进展[J].中医药临床杂志,2018,30(8):1391-1394.

[27] WAKUI S, YOKOO K, MUTO T, et al. Localization of Ang-1, -2, Tie-2, and VEGF expression at endothelial-pericyte interdigitation in rat angiogenesis. Lab Invest.2006;86(11):1172-1184.