Naoxintong Inhibits LPS Induced BV2 Cell Pyroptosis by NLRP3/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

ABSTRACT

Objective:To explore the effect of Naoxintong ethanol extract (NXT) on pyroptosis of BV2 microglia cells induced by lipopolysaccharide (LPS), and to explain the mechanism of pyroptosis based on NOD like receptor thermoprotein domain 3 (NLRP3)/cysteine-proteinase-1 (Caspase-1) pathway. Method:BV2 cells was treated with different concentrations of NXT(2, 10, 50 mg·L-1) after induced by LPS(1 mg·L-1) in vitro. Real-time PCR was used to detect mRNA expression of pro-inflammatory cytokine such as interleukin 1 beta (IL-1β), tumor necrosis factor (TNF)-α, and NLRP3.Western bolt and immunofluorescence were used to observe the protein expression of NLRP3/Caspase-1 signaling pathway. Result:Compared with control group, after LPS(1 mg·L-1) stimulation, BV2 cells viability was decreased. The mRNA expression levels of IL-1β, TNF-α and NLRP3 were significantly elevated(P<0.01), the protein levels of NLRP3 and Caspase-1 p20/Caspase-1 were also increased. After given NXT(2, 10, 50 mg·L-1), BV2 cells viability reversed which induced by LPS. Compared with LPS group, the mRNA expression of IL-1β, TNF-α and NLRP3 reduced obviously with given 50 mg·L-1NXT (P<0.05, P<0.01), significantly inhibited NLRP3 high protein expression and Caspase-1 p20/Caspase-1 expression(P<0.01). Conclusion:NXT can inhibit LPS induced pyroptosis of BV2 cells and the mechanism may closely related to NLRP3/Caspase-1 signaling pathway.