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Anion exchanger 3 (AE3) is known to serve crucial roles in maintaining intracellular chloride homeostasis by facilitating the reversible electroneutral exchange of Cl‑ for HCO3‑ across the plasma membrane. Our previous studies reported that sasanquasaponin (SQS) can inhibit hypoxia/reoxygenation (H/R)‑induced elevation of intracellular Cl‑ concentration ([Cl‑]i) and elicit cardioprotection by favoring Cl‑/HCO3‑ exchange of AE3. However, the molecular basis for SQS‑induced increase of Cl‑/HCO3‑ exchange of AE3 remains unclear. The present study demonstrated that SQS activates protein kinase Cε (PKCε) and stimulates the phosphorylation of AE3 in H9c2 cells. Notably, SQS‑induced AE3 phosphorylation was blocked by the PKCε selective inhibitor εV1‑2, and a S67A mutation of AE3, indicating that SQS could promote phosphorylation of Ser67 of AE3 via a PKCε‑dependent regulatory signaling pathway. Additionally, both inhibition of PKCε by εV1‑2 and S67A mutation of AE3 eradicated the SQS‑induced increase of AE3 activity, reversed the inhibitory effect of SQS on H/R‑induced elevation of [Cl‑]i, Ca2+ overload and generation of reactive oxygen species, and eliminated SQS‑induced cardioprotection. In conclusion, PKCε‑dependent phosphorylation of serine 67 on AE3 may be responsible for the increase of Cl‑/HCO3‑ exchange of AE3 and intracellular chloride efflux by SQS, and contributes to the cardioprotection of SQS against H/R in H9c2 cells.

molecular medicine reports

Sasanquasaponin induces increase of Cl−/HCO3− exchange of anion exchanger 3 and promotes intracellular Cl− efflux in hypoxia/reoxygenation cardiomyocytes