Astragaloside IV alleviates neuronal ferroptosis in ischemic stroke by regulating fat mass and obesity‐associated—N6‐methyladenosine—acyl‐CoA synthetase long‐chain family member 4 axis

Abstract

Ischemic stroke (IS) is a detrimental neurological disease with limited treatment options. Astragaloside IV (As-IV) was a promising bioactive constituent in the treatment of IS. However, the functional mechanism remains unclear. Here, IS cell and mouse models were established by oxygen glucose deprivation/re-oxygenation (OGD/R) and middle cerebral artery occlusion (MCAO). Quantitative reverse transcription PCR (RT-qPCR), Western blotting, or Immunofluorescence staining measured related gene and protein expression of cells or mice brain tissues, and the results revealed altered expression of acyl-CoA synthetase long-chain family member 4 (Acsl4), fat mass and obesity-associated (Fto), and activation transcription factor 3 (Atf3) after treatment with As-IV. Then, increased N 6 -methyladenosine (m 6 A) levels caused OGD/R or MCAO were reduced by As-IV according to the data from methylated RNA immunoprecipitation (MeRIP)-qPCR and dot blot assays. Moreover, through a series of functional experiments such as observing mitochondrial changes under transmission electron microscopy (TEM), evaluating cell viability by cell counting kit-8 (CCK-8), analyzing infract area of brain tissues by 2,3,5-triphenyltetrazolium chloride (TTC) staining, measuring levels of malondialdehyde (MDA), lactate dehydrogenase (LDH), Fe 2+ , solute carrier family 7 member 11 (Slc7a11) and glutathione peroxidase 4 (Gpx4) and concentration of glutathione (GSH), we found that Fto knockdown, Acsl4 overexpression or Atf3 knockdown promoted the viability of OGD/R cells, inhibited cell ferroptosis, reduced infract size, while As-IV treatment or Fto overexpression reversed these changes. In mechanism, the interplays of YTH N 6 -methyladenosine RNAbinding protein 3 (Ythdf3)/Acsl4 and Atf3/Fto were analyzed by RNA-pull down, RNA Zhenglong Jin and Wenying Gao are co-first authors.

Protective effect of vitexin against high fat-induced vascular endothelial inflammation through inhibiting trimethylamineN-oxide-mediated RNA m6A modification

“…48 In contrast, astragaloside IV promoted the transcription of FTO by upregulating ATF3, resulting in decreased m6A levels of ACSL4, thus protecting nerve cells and mice against ferroptosis-induced injury. 49 Based on the results of this study, we speculate that vitexin exerts beneficial effects on vascular inflammation by reducing serum TMAO levels and activating FTO-mediated m6A modification. Moreover, observations indicate that the binding between FTO and the small-molecule compounds is primarily driven by hydrophobic and hydrogen bonding interactions.…”

Recent advances in potential therapeutic targets of ferroptosis‑associated pathways for the treatment of stroke (Review)

“…In addition, there are studies reporting that TCM treatment can reduce the side effects of drug toxicity in patients via targeting ferroptosis, leading to significant improvements in patient safety and quality of life ( 187 , 188 ). The impact that specific Chinese medicines and their active constituents have on stroke involves multiple pathways and targets, and these are summarized in Table IV ( 125 , 182 , 189 – 191 ).…”

Mechanism of ferroptosis regulating ischemic stroke and pharmacologically inhibiting ferroptosis in treatment of ischemic stroke

“…Treatment against ischemic stroke by targeting ferroptosis has gained much attention. 192 Vitexin, 193 Angong Niuhuang Wan, 194 selenium, 195 Srs11‐92, 196 Se‐methyl L‐selenocysteine, 197 Rehmannioside A, 166 Icariside II, 198 Galangin, 199 Carvacrol, 200 Carthamin yellow, 201 β‐Caryophyllene, 202 Astragaloside IV, 203 Salvia miltiorrhiza, 204 Naotaifang extract, 133 Naotaifang, 205 Baicalein, 154 Caffeic acid, 206 Danlou tablet, 207 Rhein, 208 HSYA and AHSYB, 209 Calycosin, 159 Ferrostatin‐1, 210 Compound Tongluo Decoction, 211 Loureirin C, 212 Edaravone, 191 Quercetin, 213 Cottonseed oil, 214 DHT, 215 Dl‐3‐n‐butylphthalide, 216 Dimethyl fumarate, 217 2‐(1‐(4‐(4‐methylpiperazin‐1‐yl)phenyl)ethyl)‐10H‐phenothiazine, 218 Kaempferol, 219 Danhong injection, 220 Neutral polysaccharide from Gastrodia elata , 221 Resveratrol 190 alleviate injury after IS by inhibiting ferroptosis (Table 1 ).…”