2.2 Induction of the HPMEC under heat stress combined with LPS
in vitro
Pretreatment before molding Heat stress and lipopolysaccharide
(HS+LPS) were employed to construct the in vitro models of heat
stroke in our previous study. 15. All the HPMEC were
categorized into 4 groups with 8 wells in each group: control group (CON
group), heat stroke group (HS group), heparanase group (HPSE group), and
unfractionated heparin group (UFH group). HPSE was known to aggravate
endothelial glycocalyx degradation as the only endonuclease.16, 17 UFH was known to attenuate glycocalyx
degradation via non-covalent binding with glycocalyx.13 After conventional culture for 72 hours, the cell
in the CON group was cultured in normal conditions, without receiving
heat stress and LPS (Sigma, America). No pretreatment was performed on
the HS group before modeling. The cell in the HPSE group was subjected
to a 0.2 U/ ml Heparanase (Sigma, America) culture medium and incubated
at 37 °C for 30 minutes. The UFH group was subjected to a medium
containing 10 U/ ml unfractionated heparin (Sigma, America) and
incubated 12 hours before modeling. 18, 1920
Modeling Vitro Briefly, except for CON group, HPMEC in HS,
HPSE, and UFH groups were subjected to heat stress at 43 °C for 30
minutes, and then moved to a 5% CO2, 37 °C incubators,
stimulated with LPS at a concentration of 500 ng/ml for 12 hours.