Effect of abnormal endothelin expression in adipose-derived stem cells from obese patients on vascular endothelial cell function and its mechanism

Abudousupoer ABUDOUMING; LI Jie; MA Yun; WANG Juan

doi:10.3969/j.issn.1672-5301.2025.11.001

Abstract：Objective To clarify the abnormal expression of endothelin(EDN1)in adipose-derived stem cells(ADSCs)from obese patients through comparative analysis,and further explore its effects on vascular endothelial cell function and potential mechanism.Methods The fasting serum was collected from the obese patients who visited the Fifth Affiliated Hospital of Xinjiang Medical University from January 1,2023,to December 31,2023,and divided into the normal group(n=20)and the obese group(n=26).The levels of leptin,nitric oxide(NO)and endothelin-1(EDN1)in the serum were measured,alongside the blood pressure level taking in both the normal and obese groups.ADSCs were isolated and cultured in vitro,with purity identified by flow cytometry,differentiation ability verified by oil red O staining,and EDN1 protein expression detected by Western blot(WB).The human umbilical vein endothelial cells(HUVECs)were co-cultured with ADSCs;HUVEC proliferation was assessed using CCK-8;angiogenesis was evaluated via tube formation assay;and the expression levels of endothelial nitric oxide synthase(eNOS),EDN1,vascular endothelial growth factor A(VEGFA),serine/threonine protein kinase 1(AKT1)and phosphorylated AKT1(p-AKT1)were detected by WB.ADSCs were transfected with EDN1 knockdown/overexpression vectors and co-cultured with HUVECs after confirming transfection efficiency.CCK-8 and tube formation assays were used to test the cell proliferation and angiogenesis.RT-qPCR was used to measure the EDN1,VEGFA and AKT1 mRNA expression and WB was used to analyze the EDN1,VEGFA,AKT1/p-AKT1 and eNOS protein levels.Results Compared to the normal group,the obese group showed significantly higher blood pressure and serum levels of Leptin and EDN1(P<0.05),and significantly lower NO levels(P<0.05).ADSC analysis showed a significant upregulation of EDN1 protein in the obese group(P<0.05).Co-culture experiments indicated that,compared with the normal ADSCs group,both obese ADSCs and exogenous EDN1 overexpression significantly promoted HUVEC proliferation and angiogenesis,while inhibiting eNOS expression(P<0.05).EDN1 knockdown inhibited HUVEC proliferation and angiogenesis while promoting eNOS expression(P<0.05).EDN1 knockdown also inhibited the VEGFA/AKT1 pathway,reducing VEGFA mRNA and protein expression levels and significantly reducing the p-AKT1/AKT1 ratio(P<0.05);EDN1 overexpression produced the opposite effect(P<0.05).Conclusion The high expression of EDN1 in ADSCs from the obese patients activates the VEGFA/Akt1 pathway while inhibiting eNOS/NO activity,promoting endothelial cell proliferation and angiogenesis and contributing to the development of cardiovascular diseases.