Purification optimization of tanshinones and their neuroprotection assessment on SH-SY5Y cells injured by oxygen-glucose deprivation/reoxygenation

• Purification parameters of tanshinones were optimized by mathematical model. • Purified tanshinones raised the cell survival rate of OGD/R SH-SY5Y cells. • Purified tanshinones protected OGD/R SH-SY5Y cells by inhibiting parthanatos. To optimize purification process of tanshinones from Salvia miltiorrhiza (SM) and evaluate their neuroprotection activity against oxygen-glucose deprivation /reoxygenation (OGD/R)-induced injury in SH-SY5Y cells. Ten types of macroporous resin (MPR) were initially assessed for tanshinones adsorption capacity. Then, Plackett-Burman design (PBD) combined with response surface methodology (RSM) was applied to determine optimal purification parameters. Finally, the neuroprotective effect of purified tanshinones was investigated in OGD/R-injured SH-SY5Y cells. The highest tanshinones recovery was achieved under optimized conditons: sample volume 4 bed volumes (BV), 80% ethanol concentration, and 40 BV ethanol elution volume. The experimental comprehensive score (W) which was calculated by entropy weight method (EWM) was 0.737, closely matching the predicted value of 0.740. Furthermore, tanshinones-containing serum significantly improved ROS level, PARP-1 protein expression and AIF nuclear translocation, which are caused by OGD/R injury in SH-SY5Y cells. The integration of EWM with PBD and RSM provided an effective framework for multi-component purification optimization. Purified tanshinones demonstrated potent inhibition of parthanatos in OGD/R-injured neuronal cells. This study not only establishes a scalable methodology for plant-derived compound purification but also proposes novel therapeutic strategies for cerebral ischemia and related neurological disorders.