Fructus Gardenia Extract Improves Cerebral Ischemia-Reperfusion Injury via RIPK1-RIPK3-MLKL Axis

Fructus Gardenia (FG), which was riched in crocin, showed protective effects in rats with middle cerebral artery occlusion (MCAO). This article attempts to explore the new therapeutic effect of FG from the perspective of necroptosis. The pathological process of cerebral ischemia-reperfusion injury (CIRI) was simulated in vivo and in vitro in the MCAO rat and oxygen glucose deprivation reperfusion (OGD/R) cell models. After FG treatment, the neurological deficit score, TTC staining, TUNEL staining and western blot were used to evaluate the nerve injury, cerebral infarction volume and brain tissue necroptosis. After in vitro drug treatment, cell viability, necrosis, drug toxicity and typical signaling pathways of necroptosis were tested by CCK-8, flow cytometry, lactate dehydrogenase (LDH) release assay and western blot. FG treatment significantly improved neurological dysfunction caused by MCAO treatment in rats, reduced brain infarct volume and necrosis, and prevented abnormal activation of the necroptosis classic pathway RIPK1-RIPK3-MLKL. In vitro experiments, FG enhanced the viability of model cells, reduced OGD/R-induced cell necrosis and LDH release, and down-regulated protein expression of RIPK1-RIPK3-MLKL. Necrostatin-1, a necrotrophic apoptosis inhibitor, enhanced the effects of FG. FG extract reduced necroptosis in brain tissue cells and alleviated CIRI in rats, RIPK1-RIPK3-MLKL axis may be involved in this process.