Neuroprotective Effects of Rg1 against H2O2

Primary hippocampal neurons were prepared from postnatal Sprague-Dawley rats (within 24 h; 6-8 neonates/experiment) obtained from the Center of Laboratory Animals of Anhui Medical University (Hefei, China) and placed in plates coated with poly-L-lysine (10 µg/ml). Neurobasal medium with B-27 supplement (Thermo Fisher Scientific, Inc., Waltham, MA, USA) was used to culture the neurons at 37°C with 5% CO₂ as described previously (21 (link)). The neurons were cultured for 7 days and divided into six groups: Control group, H₂O₂ (200 µM) group, H₂O₂ (200 µM) + Tempol (100 µM) group, and H₂O₂ (200 µM) + Rg1 (1, 5 and 10 µM) groups. With the exception of the control group, the neurons in the groups were treated with H₂O₂ (200 µM) and Tempol (100 µM) or Rg1 (1, 5 or 10 µM) for 24 h. Tempol (Santa Cruz Technology, Inc., Dallas, TX, USA) and Rg1 (content of Rg1 >98%; Chengdu Desite Biotechnology Co., Chengdu, China) were dissolved in distilled water and stored at −80°C. All experimental procedures were performed in accordance with the approved protocol of the Ethics Committee of Anhui Medical University (Anhui, China).