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. 2021 Apr 16;43:101977. doi: 10.1016/j.redox.2021.101977

Fig. 5.

Fig. 5

Compound 23 directly reacts with GSH. (A, B, C, D) HepG2 cells were treated with 1.25, 2.5 and 5 μM 23 for 24 h or 2.5 μM 23 for 0, 3, 6, 12, 24 h, and the levels of GSH (A, C) and the ratio of GSH/GSSG (B, D) were analyzed using GSH detection kits. *P < 0.05, **P < 0.01, relative to control or 0 h. (E) Scheme for synthesis of 23-GSH. (F) The 23-GSH was detected by UPLC-MS/MS. Standard substances (a); Control group (b); 23 treated group (c); Ion extraction of 23-GSH in the 23 treated group (d). After treatment with 0 or 2.5 μM 23 for 8 h, the cells were collected and lysed using RIPA lysis buffer. (G) The reaction was monitored by the change of representative hydrogen signals with 1H NMR analysis. Compound 23 (1.2 mg) were incubated with an excess of GSH (2.4 mg) in deuterated phosphate buffer at pH 7.4 (0.02 mL) and DMSO‑d6 (0.50 mL) at 37 °C. Results are expressed as mean ± SD, (n = 3).