Effects and Action Mechanism of Huoxue Tongluo Formula on the Formation of Neutrophil Extracellular Traps

Figure 4

Effect of HXTLF on the formation mechanism of NETs. (a) Western blot detection of the expression of AKT1, p-AKT1, IKK, p-IKK, NF-κB, and p-NF-κB in neutrophils (left). β-Actin was employed as an internal reference protein. Semiquantitative analysis was used for protein quantification and ImageJ was utilized to analyze the gray value of protein bands. The results were displayed in column diagram (right). (b) Immunofluorescence microscopy of H3Cit and MPO, and the right was mean fluorescence intensity calculated. Magnification 400 ×. Scale bars = 50 µm. (c) NETs cell free DNA detection. (d) Detection of secretion factor IL-1β in neutrophil supernatant. Activator (pretreatment with SC79 in 8 µg/ml for 30 min); Inhibitor (pretreatment with MK-2206 2HCl in 10 µm/L for 30 min). DMSO was used as the activator and inhibitor control. The data were obtained from three independent experiments and expressed as mean ± SD. Asterisks indicated significance: ; compared to control, n = 3.

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