Matrine protects PC12 cells from lipopolysaccharide-evoked inflammatory injury via upregulation of miR-9

Abstract

Context Matrine is a well-known anti-inflammatory quinolizidine alkaloid derived from leguminous plant Sophora flavescens Ait. (Leguminosae). Objective This study was designed to uncover the potential application of matrine in treating spinal cord injury (SCI). Materials and methods Neuron-like PC12 cells in experimental groups were pre-treated with/without matrine (200 μM) for 24 h and then stimulated by lipopolysaccharide (LPS, 5 μg/mL) for 12 h. PC12 cells in control group were cultured in complete medium. CCK-8 assay, flow cytometry, qRT-PCR, western blot and ELISA were performed to evaluate cell damage. Moreover, after cells were transfected with miR-9 inhibitor for 48 h, above indicators were tested again. qRT-PCR and western blot were also conducted to uncover the downstream effectors and signalling pathways for matrine. Results LPS (5 μg/mL) decreased cell viability about 50%. Matrine (200 μM) decreased cell viability about 0%, 13.8% and 30% at 24 h, 48 h and 72 h, respectively. The loss of viability, stimulation of apoptosis, and release of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) evoked by LPS were attenuated by the pre-treatment of matrine partly. Meanwhile, LPS reduced miR-9 expression about 60%, but matrine completely reversed LPS-decreased miR-9 level. By silencing miR-9 expression, the protective properties of matrine towards PC12 cells were impeded. Besides, matrine inhibited the activation of JNK and NF–κB pathways even under the condition of LPS. And the impact of matrine on the signalling were attenuated by miR-9 silencing. Discussion and Conclusion This paper provided in vitro evidence that matrine was able to protect PC12 cells against LPS-evoked damage. The neuroprotective properties of matrine may be due to its regulation of miR-9 expression as well as JNK and NF–κB pathways.