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

Abstract

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-κB (NF-κB) 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-κB and JAK/STAT signaling pathways. We conclude that HPMSCs/leptin could promote angiogenic potential and peripheral inflammation of HUVECs after X-ray radiation.

Radiobiological Studies of Microvascular Damage through In Vitro Models: A Methodological Perspective

“…EC migration is analyzed in vitro to evaluate interactions with other cells or stimuli and study possible pathways involved in angiogenesis during tissue revascularizations (e.g., wound healing). The most common method to study this phenomenon involved migration assays (23 studies, 15.6%) [ 33 , 34 , 35 , 37 , 39 , 57 , 58 , 62 , 64 , 76 , 84 , 85 , 94 , 97 , 107 , 109 , 110 , 119 , 123 , 146 , 147 , 148 , 149 ]. Briefly, in such assay, cells were seeded on a porous membrane, and a chemotactic stimulus is then added on the other membrane side ( Figure 4 e).…”

“…Inflammation was mainly addressed by quantifying cytokines released in culture medium through ELISA assays or specific kits (e.g., intracellular ROS kit). The most commonly studied cytokines were interleukins (IL), analyzed in 20 studies (13.6%) [ 21 , 23 , 29 , 32 , 46 , 48 , 49 , 50 , 53 , 69 , 99 , 126 , 145 , 146 , 147 , 150 , 151 , 152 , 153 , 154 ]. IR application to ECs increased production of specific cytokines such as IL-1α, mediating bystander effects [ 99 ], and upregulated pro-inflammatory cytokines and chemokines (including IL-6, IL-1α, and MCP-1), indicators of cell senescence [ 126 ].…”

“…The presence of other cytokines may also influence IL production, as seen in [ 29 ], in which IL-4 and IL-10 affected IL-6 and IL-8 production due to radiation. Possible biological pathways involved in IL production after irradiation have been proposed, including p38/MAPK and NF-κB [ 50 ] or thrombomodulin [ 49 , 50 , 145 , 146 ], also evaluating their possible inhibition through different agents ([ 49 , 145 , 146 , 154 ]). p38 pathway was also involved in ECs bystander effect: irradiated macrophages increased NO production, activating the p38 pathway in ECs, leading to increased apoptosis and inflammatory response [ 63 ].…”

Knockdown of lnc-KCNC3-3:1 Alleviates the Development of Atherosclerosis via Downregulation of JAK1/STAT3 Signaling Pathway

“…In addition, it is well known that endothelial cell injury is the early event during the occurrence of atherosclerosis (1). In other words, atherosclerosis is a chronic inflammatory response of the arterial wall to endothelial cell damage (21,22). For example, the expression of proteins associated with the JAK/STAT signaling pathway is notably upregulated in patients with atherosclerosis (23).…”

Adiponectin and Leptin Exert Antagonizing Effects on HUVEC Tube Formation and Migration Modulating the Expression of CXCL1, VEGF, MMP-2 and MMP-9

“…In addition, it has been proved that induction of FGF2 as well as of the matrix metalloproteinase-2 (MMP-2) and MMP-9 signaling pathways are necessary for leptin angiogenesis [ 25 , 27 ]. However, it is to notice that most of the data suggest a role for leptin in mediating cancer aberrant angiogenesis [ 28 ].…”