[Retracted] Chrysophanol Ameliorates Hemin-Induced Oxidative Stress and Endoplasmic Reticulum Stress by Regulating MicroRNA-320-5p/Wnt3a Pathway in HT22 Cells

Figure 4

CHR activated the Wnt3a/β-catenin signaling pathway by downregulating miR-320-5p in hemin-induced HT22 cell injury models. (a) Potential binding site of miR-320-5p at the 3-UTR of Wnt3a mRNA is predicted using the miRWalk software. The red color indicates wild-type, and the green color indicates the mutated sequence of Wnt3a at the miR-320-5p-binding site. (b) HT22 cells were cotransfected with miR-320-5p (or NC mimics) and a pmir-GLO vector containing wild-type or mutated miR-320-5p-binding sites (mutated) at Wnt3a 3-UTR. Luciferase reporter assays were performed 24 h after cotransfection. NC mim.: negative control mimics; miR mim.: miR-320-5p mimics; NC inh.: negative control inhibitor; miR inh.: miR-320-5p inhibitor. ^N.S., . . (c) HT22 cells were transfected with the miR-320-5p inhibitor or miR-320-5p mimics and their corresponding controls. The expression of miR-320-5p and Wnt3a mRNA was analyzed using the RT-PCR technique. . . (d and e) Expression levels of mRNA and protein of Wnt3a were analyzed in hemin-induced HT22 cells with or without CHR (10 μM; treatment time: 24 h) using the RT-PCR and western blot. and . . (f) Expression of miR-320-5p was analyzed in hemin-induced HT22 cells in the presence and absence of CHR (10 μM; treatment time: 24 h) or miR-320-5p mimics (treatment time: 24 h) using the RT-PCR technique. . . (g) The mRNA expression of Wnt3a was analyzed in hemin-induced HT22 cells with or without CHR (10 μM; treatment time: 24 h) or miR-320-5p mimics (treatment time: 24 h) using the RT-PCR technique. . . (h) The protein expression levels of Wnt3a, total β-catenin (T-β-catenin), and nuclear β-catenin (N-β-catenin) were analyzed in hemin-induced HT22 cells with or without CHR (10 μM; treatment time: 24 h) or miR-320-5p mimics (treatment time: 24 h) using the western blot technique. , , and . .

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