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| Effect of Glaucocalyxin A on Growth and Biofilm Formation of Mycobacterium Abscessus via Regulating TLR4/NF-κB Signaling Pathway |
| LI Qian, REN Zhe, YANG Bingzhou, et al |
| Hebei Chest Hospital/Hebei Provincial Key Laboratory of Lung Disease, Hebei Shijiazhuang 050000, China |
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Abstract Objective: To investigate the effects of glaucocalyxin A (GLA) on on the growth and biofilm formation of Mycobacterium abscessus via regulating toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway. Methods: Mycobacterium abscessus strain ATCC19977 was inducted into blank group, GLA low, medium high-dose group, TAK-242 group and GLA high-dose group + LPS group. The lowest drug concentration (MIC90) corresponding to an inhibition rate of 90% or greater on ATCC19977 by GLA determined by Alma Blue liquid drug sensitivity assay. The biofilm formation in ATCC19977 was recorded by scanning electron microscopy. Crystal violet staining was employed to detect the total biomass of ATCC19977 biofilm, MTT assay was used to detect the metabolic activity of ATCC19977 biofilm. The quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect the mRNA expression of MAB-2030 and MAB-2028 and the proteins expression of TLR4 and p-NF-κB p65 in ATCC19977. Results: The MIC90 of GLA on ATCC19977 was 10μmoL/L. Compared with the blank group, the GLA dose groups showed a decrease in the number of ATCC19977, and the biofilm formation was damaged, manifested as a thinner thickness and a loose state. The total biomass and metabolic activity of ATCC19977 biofilm, mRNA expression of MAB-2030 and MAB-2028, protein expression of TLR4 and p-NF -κB p65 in ATCC19977 of the GLA dose groups decreased significantly, and the GLA high-dose group showed the most obvious trend, the corresponding indicator change trend in TAK-242 group was consistent with the above (all P<0.05). Compared with the GLA high-dose group, the GLA high-dose+LPS group had an increase in the number of ATCC19977, a decrease in the degree of biofilm damage, increases in total biofilm biomass, biofilm metabolic activity, MAB-2030 and MAB-2028 mRNA expression, and TLR4 and p-NF -κB p65 protein expression in ATCC19977 (all P<0.05). Conclusion: GLA may inhibit the growth and biofilm formation of ATCC19977 by suppressing the TLR4/NF -κB signaling pathway.
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2025, Vol. 31 
