Date of Award

2025

Document Type

Dissertation

Degree Name

Pharmaceutical Sciences (Ph.D.)

Department

Pharmaceutical Sciences

First Advisor

Francis A.X. Schanne

Second Advisor

Xingguo Cheng

Abstract

Exosomes are membrane bound subcellular vesicles formed through membrane budding of a cell and play important roles in cell-cell communication. Studies have demonstrated inflammatory changes in various cell types depending on the etiology of the exosomes such as exosomes from cancer cells or exosomes derived from mesenchymal stem cells. The purpose of this study was to determine the inflammatory consequences of exosomes in microglial cells following exposure to lead, lipopolysaccharides (LPS) and lead+LPS. The cultured mouse microglial cells were treated at 0.01 μM, 0.1 μM and 1 μM of lead,- three environmentally relevant levels and LPS was used at 10 pg/mL in all LPS samples. Inflammation was assessed by determining the levels of inflammatory markers of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), interleukin 10 (IL-10) and toll-like receptor 4 (TLR-4). Analysis was conducted with quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) methods. Exosomes derived from healthy microglia cells were grown and split. Each biological sample had exosomes generated from an individual flask. 20 ng/mL of exosomes were added to microglia with lead and LPS. Cell lysate and supernatant from immortalized microglia were exposed for eighteen hours to 0.01 μM of lead simultaneously exposed to LPS led to apparent lower levels of cytokines compared to LPS alone, while 1 μM of lead with LPS showed higher cytokine levels than microglia at 0.01 μM of lead with LPS.

Download

Included in

Toxicology Commons

Share

COinS