Hypertonic saline protects brain endothelial cells against hypoxia correlated to the levels of estimated glomerular filtration rate and interleukin-1β

Abstract

Objective:The aim of this study was to verify the protective effect of hypertonic saline (HS) on brain endothelial cells under hypoxic conditions and the relevant underlying mechanism.Methods:bEnd.3 cells were treated with oxygen-glucose deprivation (OGD)-induced injury. To measure HS performance, cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium salt assay, and cell apoptosis was assessed by flow cytometry and Terminal deoxynucleotidyl transferase UTP nick-end labeling staining. RNA-seq was performed to assess the expression profiles and screen the candidate genes that participated in OGD-induced injury and the HS protective effect. Quantitative real-time polymerase chain reaction (qPCR) and western blot analysis were used to confirm the expression of candidate genes, and enzyme-linked immunosorbent assay was used to measure the level of interleukin (IL)-1β. Overexpression analyses were performed to confirm the functions of the differentially expressed genes.Results:HS with a concentration of 40 mmol/L NaCl had an obvious protective effect on bEnd.3 cells after OGD-induced injury, resulting in increased cell viability and a smaller percentage of apoptotic cells. According to the RNA-seq results, epidermal growth factor receptor (EGFR) was chosen as the differentially expressed gene target in this study. The qPCR and western blot analyses further confirmed that the levels of EGFR/phosphorylated epidermal growth factor receptor and IL-1β were enhanced after OGD-induced injury, but attenuated after treatment with 40 mmol/L of NaCl HS. Overexpressed EGFR reversed the protective effect of HS that caused low viability and high rates of apoptosis in cells.Conclusion:HS can protect endothelial cells against OGD-induced injury, but is affected by the expression of EGFR/p-EGFR and IL-1β.

The impact of endothelial cell death in the brain and its role after stroke: A systematic review

“…In bEnd.3 cells, Liu and colleagues found a time-dependent increase in the levels of cleaved caspases-3 and -9 between 6 and 12 hours of OGD [91]. Other studies corroborate these findings of apoptotic cell death in bEnd.3 cells [92–96], in a human brain microvascular EC line (HBMEC [97–99]) as well as in primary brain microvascular ECs from rats [100, 101] and mice [75, 102–104].…”

Blood-Brain Barrier Dysfunction Amplifies the Development of Neuroinflammation: Understanding of Cellular Events in Brain Microvascular Endothelial Cells for Prevention and Treatment of BBB Dysfunction

“…Although it is not completely understood whether all of these BMEC-derived soluble factors induced by IL-1β disrupt the barrier integrity of BMECs, these factors may participate in the destructive effect of IL-1β on the BBB in an autocrine manner. Importantly, the ability of BMECs to secrete IL-1β in response to oxygen glucose deprivation and TNF-α should also be considered in IL-1β-induced BBB dysfunction ( Matsumoto et al, 2014 ; Chen et al, 2017 ).…”

Quantitative Investigation into the influence of intravenous fluids on human immune and cancer cell lines

“… Flow cytometry, RNA-seq, Quantitative real-time polymerase chain reaction, western blot, enzyme-linked immunosorbent assay How do the levels of IL-1β and EGFR is correlated to the performance of HTS and apoptosis? 41 Hypertonic saline (HTS) HeLa cells Increasing NaCl concentration more than 130 mM decreases growth rate, increases protein levels, and cell volume. Biochemical analysis, hemocytometry, microscopy Ionic and osmotic effects of HTS in the metabolism of HeLa cells 50 Hypertonic sucrose (HTSu) Human leukemia cells(U937) U937 cells developed apoptosis thanks to regulatory volume increase in HTSu.…”

“…Most IVF investigations have focused on revealing their influence on medication stability, compatibility and administration 18 . To date, there have been few quantitative in vitro studies about the influence of IVF on cellular level, which are mostly reported for blood cells and endothelial cells 18,40,41 .…”