Tectorigenin protects cell damage in Parkinson's disease model by regulating miR-103a-3p/SHC3 pathway

Abstract

Objective To investigate the effect of tectorigenin on cell model of Parkinson's disease (PD), so as to further analyze whether its mechanism is related to the regulation of miR-103a-3p/SHC adaptor protein 3 (SHC3) pathway. Methods PC12 cells were treated with 250 mmol/L 1-methyl-4-phenylpyridine ion (MPP+) to establish a PD cell model. Then, the cells were divided into control group and experimental groups as follows: PD group, PD+50 μmol/L tectorigenin, PD+100 μmol/L tectorigenin, PD+200 μmol/L tectorigenin, PD+anti-miR-NC, PD+miR-103a-3p inhibitor, and PD+tectorigenin+miR-103a-3p mimic. Cell viability and apoptotic rate of each group were evaluated using CCK-8 assay and flow cytometry respectively. The activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) and release of lactate dehydrogenase (LDH) of cells in each group were subsequently detected. Moreover, RT-qPCR and Western blotting were performed to determine the mRNA expression of miR-103a-3p and protein level of SHC3, respectively, and dual luciferase reporting assay was adopted to analyze the targeting relationship between miR-103a-3p and SHC3. Results As compared with the control group, the viability, SOD activity, and SHC3 expression of cells were notably reduced (P<0.05), while the apoptotic rate, MDA content, LDH release, and miR-103a-3p expression were significantly increased (P<0.05) in the PD group. When compared with the PD group, the viability, SOD activity, and SHC3 expression of cells in the PD+tectorigenin (50, 100 and 200 μmol/L) groups were all notably improved (P<0.05), while the apoptotic rate, MDA content, LDH release and miR-103a-3p level were greatly declined (P<0.05). In addition, the cell viability, SOD activity and SHC3 level were higher, whereas the apoptotic rate, MDA content and LDH release were lower in the PD+miR-103a-3p inhibitor group than in the PD+anti-miR-NC group (P<0.05). Finally, the cell viability, SOD activity and SHC3 expression in the PD+tectorigenin+miR-103a-3p mimic group were notably decreased (P<0.05), while the apoptotic rate, MDA content, and LDH release were increased (P<0.05), as compared with the PD+tectorigenin groups. Dual luciferase reporting assay showed that SHC3 was a direct target of miR-103a-3p. Conclusion Tectorigenin protects PD model cells against apoptosis and oxidative damage, which may be related to the inhibition of miR-103a-3p/SHC3 pathway.

Keywords

• tectorigenin
• parkinson's disease
• mir-103a-3p
• shc adaptor protein 3
• apoptosis
• oxidative stress