Inhibitory Effect of Aconiti Radix Water Extract and Its Monomeric Components on the Secretion of α-Hemolysin in Staphylococcus Aureus
Objective To investigate the inhibitory effect of Aconiti Radix water extract and its monomeric components on the secretion of α-hemolysin in Staphylococcus aureus.Methods RNAseq was used to analyze the effect of Aconiti Radix water extract on the transcription levels of USA300 hla,psma,and isda genes in Staphylococcus aureus.The reverse transcription real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)method was used to determine the effect of Aconiti Radix water extract and its monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL)on the expression levels of hla,psma,and isda genes.The quantitative hemolysis test was used to evaluate the hemolytic activity.Western blot was used to investigate the effect of monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL)on the secretion of α-hemolysin in the supernatant of Staphylococcus aureus bacterial solution.Results After USA300 was treated with Aconiti Radix water extract(250 mg/mL),genes related to virulence factors such as α-hemolysin,δ-hemolysin,and phenol-soluble regulatory proteins were significantly down-regulated(P<0.005).After USA300 was treated with the Aconiti Radix water extract(250 mg/mL)and its monomeric components of aconitine(1 mg/mL),hypaconitine(1 mg/mL),and mesaconitine(1 mg/mL),the transcript levels of hla,psma,and isda genes were significantly inhibited(P<0.05),with the most significant inhibition level of the hla gene.The hemolytic activity of Staphylococcus aureus was significantly inhibited(P<0.01).After USA300 was treated with the monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL),the secretion of α-hemolysin in the supernatant of Staphylococcus aureus decreased in a concentration dependent manner with aconitine,but the trend was not significant after treatment with monomeric components of hypaconitine and mesaconitine.Conclusion The Aconiti Radix water extract and its monomeric components can inhibit the secretion of α-hemolysin in Staphylococcus aureus,especially the monomeric component aconitine has the potential to be used for anti-Staphylococcus aureus infection.
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