Objective:

2

To observe the protective effect and mechanism of hydroxyl safflower yellow A (HSYA)

Methods:

2

HUVECs were treated with oxygen-glucose deprivation reperfusion (OGD/R) to simulate the ischemia reperfusion model

and cell counting kit-8 was used to detect the protective effect of different concentrations (1.25–160 μmol/L) of HSYA on HUVECs after OGD/R. HSYA 80 μmol/L was used for follow-up experiments. The contents of inflammatory cytokines interleukin (IL)-18

IL-1β

monocyte chemotactic protein 1 (MCP-1)

tumor necrosis factor α (TNF-α) and IL-6 before and after administration were measured by enzyme-linked immunosorbent assay. The protein expressions of toll-like receptor

NOD-like receptor containing pyrin domain 3 (NLRP3)

gasdermin D (GSDMD) and GSDMD-N-terminal domain (GSDMD-N) before and after administration were detected by Western blot. NLRP3 inflammasome inhibitor cytokine release inhibitory drug 3 sodium salt (CRID3 sodium salt

also known as MCC950) and agonist were added

and the changes of NLRP3

cysteine-aspartic acid protease 1 (Caspase-1)

GSDMD and GSDMD-N protein expressions were detected by Western blot.

Results:

2

HSYA inhibited OGD/R-induced inflammation and significantly decreased the contents of inflammatory cytokines IL-18

IL-1β

MCP-1

TNF-

α and IL-6 (

P

<

0.01 or

P

<

0.05). At the same time

by inhibiting NLRP3/Caspase-1/GSDMD pathway

HSYA can reduce the occurrence of pyroptosis after OGD/R and reduce the expression of NLRP3

Caspase-1

GSDMD and GSDMD-N proteins (

P

<

0.01).

Conclusions:

2

The protective effect of HSYA on HUVECs after OGD/R is related to down-regulating the expression of NLRP3 inflammasome and inhibiting pyroptosis.