Hispidin Inhibits Ferroptosis Induced by High Glucose via the miR-15b-5p/GLS2 Axis in Pancreatic Beta Cells

Figure 4

MiR-15b-5p directly targets GLS2 to inhibit ferroptosis induced by HG levels in PBCs. (a) The binding sites between the 3′ UTR of GLS2 and miR-15b-5p have been illustrated (left), and the interaction between miR-15b-5p and the 3′ UTR of GLS2 was verified by the dual-luciferase activity assay. (b) PBCs transfected with miR-NC or miR-15b-5p mimic for 24 h; the mRNA levels of GLS2 were measured by RT-PCR. The untreated PBCS was used as the control. (c) PBCs transfected with miR-NC or miR-15b-5p mimic for 24 h; the protein levels of GLS2 were measured by Western blot. The untreated PBCs were used as the control. (d) PBCs transfected with siRNAs against GLS2 for 24 h; the mRNA levels of GLS2 were measured by RT-PCR. (e) PBCs transfected with siRNAs against GLS2 for 24 h; protein levels of GLS2 were measured by Western blot. (f)–(k) Under the HG condition, PBCs were transfected with different siRNAs for 24 h following which the cells were treated with hispidin for an additional 24 h. Cells cultured under normal conditions were used as the control. (f) Cell viability measured. (g) Cell death measured. (h) ROS levels were measured. (i) Fe²⁺ levels were measured. (j) MDA levels measured. (k) GSH levels measured. Data are representative of at least three independent experiments ().

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)