EPCs-Exo inhibit paclitaxel-induced pyrolysis of vascular endothelial cells
WANG Ping1, MENG Liping1,LIU Longbin2, PENG Fang1
1.Department of Cardiology, Shaoxing People’s Hospital (Shaoxing Hospital of Zhejiang University), Shaoxing 312000, China; 2.Department of Cardiology, Affiliated Hospital of Shaoxing College of Arts and Sciences (Shaoxing Municipal Hospital), Shaoxing 312000, China
WANG Ping,MENG Liping,LIU Longbin, et al. EPCs-Exo inhibit paclitaxel-induced pyrolysis of vascular endothelial cells[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2022, 52(1): 9-14.
Abstract：Objective: To explore the effect of endothelial progenitor cell exosomes (EPCs-Exo) on paclitaxel-induced endothelial cell pyrolysis and its possible mechanism. Methods: The primary bone marrow mesenchymal stem cells (BMSCs) and ECPs were cultured and identified. The exosomes in the cultured medium of BMSCs and ECPs were extracted, and the morphology of the exosomes was observed by transmission electron microscope. The expression of CD63 and CD9 was detected by western blot to identify the exosomes. BMSCs-Exo and EPCs-Exo were used to intervene the endothelial cell pyrolysis model induced by paclitaxel, and the cell morphology was observed by microscope; Western blot was used to detect the exosomal marker proteins CD63,CD9 and pyrolysis-related proteins NLRP-3, IL-18, GSDMD-N. The ultrastructure change of the cells were observed under electron microscope. RNA sequencing was used to detect the difference of non-coding RNA in the two groups of exosomes, and qRT-PCR was performed to detect the expression of LncRNA FGD5-AS1 in each group. Results: BMSCs and EPCs were isolated, cultured and differentiated from rat bone marrow cells. The positiveness of CD90, CD44 in BMSCs and CD34, VEGFR2 in EPCs were confirmed by immunofluorescence.Compared with the control group, the cell membrane of the paclitaxel group was destroyed, the distance between mitochondria in the cell increased, the arrangement of intracellular fibers was disordered, and the expression of pyrooptosis proteins such as NLRP-3, IL-18, GSDMD-N was increased (P<0.05). Compared with the paclitaxel group, the cell membrane of the paclitaxel+EPCs-Exo group was more complete, the cell line damage was mild, and the expression of NLRP-3, IL-18, and GSDMD-N was reduced (P<0.05). The microarray and RT-PCR results showed that compared with BMSCs-Exo, the expression of LncRNA FGD5-AS1 in EPCs-Exo increased (P<0.05). Conclusion: EPCs-Exo inhibited paclitaxel-induced pyrolysis of vascular endothelial cells.
