关键词: 体外冲击波, 内皮-间充质转化, 糖尿病血管病变, 锌指转录因子, 丝裂原活化蛋白激酶

Abstract:

Objective To investigate the effect of extracorporeal shock wave therapy （ESWT） on high glucose-induced endothelial?mesenchymal transition （EndMT） in human aortic endothelial cells （HAECs） and explore its potential mechanism. Methods HAECs were cultured in a high?glucose medium to establish an EndMT model. The cells were divided into five distinct groups： the normal control group （CON）， the control+shockwave group （CE）， the high?glucose group （HG）， the high?glucose + ESWT group （ESWT）， and the Snail inhibitor group （CYD19）. In the CON and CE groups， the glucose concentration was set at 5 mmol/L， while in the HG， ESWT， and CYD19 groups， it was 25 mmol/L. After culturing the cells for 7 days， the cells in the intervention groups were resuspended and subjected to shockwave treatment （8 Hz， 0.1 mJ/mm2， 1 000 pulses） once every 2 days， with a total of 3 treatments. Calcium deposition was detected using Alizarin Red S staining. The expression levels of endothelial markers （CD31， VEGF） and mesenchymal markers （α?SMA， FSP?1， OPG） were determined through Western blot and immunofluorescence analyses. Additionally， the expression of Snail， along with the total and phosphorylated levels of ERK1/2 and p38， were also measured. Results Alizarin Red S staining demonstrated that calcium deposition was significantly elevated in the HG group when compared with the CON group， and ESWT intervention notably reduced calcium deposition. In comparison with the CON group， the HG group showed a significant decrease in the expression of CD31 and VEGF， but an increase in the expression of α?SMA， FSP?1， and OPG. Meanwhile， the total and phosphorylated levels of ERK1/2 and p38， along with Snail expression， were significantly upregulated. When compared with the HG group， ESWT intervention significantly upregulated the expression of CD31 and VEGF， downregulated the expression of α?SMA， FSP?1， and OPG， inhibited the expression and phosphorylation of ERK1/2 and p38， and reduced the levels of Snail protein. Moreover， Snail expression was significantly decreased in both the ESWT and CYD19 groups as compared with the HG group. Conclusions The biological effects mediated by ESWT can effectively ameliorate the calcification of HAECs in a high?glucose environment. The underlying mechanism is likely that ESWT downregulates the expression and phosphorylation of p38 and ERK1/2 in the MAPK signaling pathway， thus inhibiting the expression of the downstream transcription factor Snail and reversing EndMT.

Key words: extracorporeal shock wave, endothelial-mesenchymal transition, diabetic angiopathy, snail, mitogen?activated protein kinase