Effects of leptin-modified human placenta-derived mesenchymal stem cells on angiogenic potential and peripheral inflammation of human umbilical vein endothelial cells (HUVECs) after X-ray radiation

• Published in: Journal of Zhejiang University. B. Science 2020-04, Vol.21 (4), p.327-340
• Main Authors: Chen, Shu, Wang, Qian, Han, Bing, Wu, Jia, Liu, Ding-kun, Zou, Jun-dong, Wang, Mi, Liu, Zhi-hui
• Format: Article
• Language: English
• Subjects: Angiogenesis; Assaying; Biomedical and Life Sciences; Biomedicine; Blood vessels; Cell culture; Cell growth; Cell migration; Cell Proliferation; Cells, Cultured; Cholecystokinin; Colony-stimulating factor; Culture; Cytokines; Cytokines - biosynthesis; Endothelial cells; Enzyme-linked immunosorbent assay; Female; Fibroblast growth factor 2; Granulocyte-macrophage colony-stimulating factor; Growth factors; Human Umbilical Vein Endothelial Cells - physiology; Human Umbilical Vein Endothelial Cells - radiation effects; Humans; Inflammation; Inflammation - etiology; Interleukin 6; Interleukin 8; Kinases; Leptin - pharmacology; Leukocytes (granulocytic); Macrophages; Mesenchymal stem cells; Mesenchymal Stem Cells - physiology; Neovascularization, Physiologic - physiology; Placenta; Placenta - cytology; Polymerase chain reaction; Pregnancy; Radiation damage; Signal transduction; Signaling; STAT3 Transcription Factor - genetics; Stem cells; Transcription; Transcription Factor RelA - genetics; Vascular endothelial growth factor; Veins; X-Rays
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.B1900598

Combined radiation-wound injury (CRWI) is characterized by blood vessel damage and pro-inflammatory cytokine deficiency. Studies have identified that the direct application of leptin plays a significant role in angiogenesis and inflammation. We established a sustained and stable leptin expression system to study the mechanism. A lentivirus method was employed to explore the angiogenic potential and peripheral inflammation of irradiated human umbilical vein endothelial cells (HUVECs). Leptin was transfected into human placenta-derived mesenchymal stem cells (HPMSCs) with lentiviral vectors. HUVECs were irradiated by X-ray at a single dose of 20 Gy. Transwell migration assay was performed to assess the migration of irradiated HUVECs. Based on the Transwell systems, co-culture systems of HPMSCs and irradiated HUVECs were established. Cell proliferation was measured by cell counting kit-8 (CCK-8) assay. The secretion of pro-inflammatory cytokines (human granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-1α, IL-6, and IL-8) was detected by enzyme-linked immunosorbent assay (ELISA). The expression of pro-angiogenic factors (vascular endothelial growth factor ( VEGF ) and basic fibroblast growth factor ( bFGF )) mRNA was detected by real-time quantitative polymerase chain reaction (RT-qPCR) assay. Relevant molecules of the nuclear factor-KB (NF- k B) and Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathways were detected by western blot assay. Results showed that leptin-modified HPMSCs (HPMSCs/leptin) exhibited better cell proliferation, migration, and angiogenic potential (expressed more VEGF and bFGF ). In both the single HPMSCs/leptin and the co-culture systems of HPMSCs/leptin and irradiated HUVECs, the increased secretion of pro-inflammatory cytokines (human GM-CSF, IL-1α, and IL-6) was associated with the interaction of the NF- K B and JAK/STAT signaling pathways. We conclude that HPMSCs/leptin could promote angiogenic potential and peripheral inflammation of HUVECs after X-ray radiation.