AUTHOR=Zhang Wenxia , He Min , Kong Nana , Niu Yuxiao , Li Anhong , Yan Yuzhong 

TITLE=Study on the inhibition activity and mechanism of Tanreqing against Klebsiella pneumoniae biofilm formation in vitro and in vivo

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2024.1368450

DOI=10.3389/fcimb.2024.1368450

ISSN=2235-2988

ABSTRACT=<sec><title>Objective</title><p>To evaluate the antibacterial effect of Tanreqing (TRQ) against <italic>K. pneumoniae</italic> and its inhibition activity on bacterial biofilm formation <italic>in vitro</italic> and <italic>in vivo</italic>, and to explore the mechanism of the inhibitory effects of TRQ on <italic>K. pneumoniae</italic> biofilm formation.</p></sec><sec><title>Methods</title><p>An <italic>in vitro</italic> biofilm model of <italic>K. pneumoniae</italic> was established, and the impact of TRQ on biofilm formation was evaluated using crystal violet staining and scanning electron microscopy (SEM). Furthermore, the clearance effect of TRQ against <italic>K. pneumoniae</italic> in the biofilm was assessed using the viable plate counting method; q-RT PCR <italic>w</italic>as used to evaluate the inhibitory effect of different concentrations of TRQ on the expression of biofilm-related genes in <italic>Klebsiella pneumoniae</italic>; The activity of quorum sensing signal molecule AI-2 was detected by <italic>Vibrio harveyi</italic> bioluminescence assay; Meanwhile, a guinea pig lung infection model of <italic>Klebsiella pneumoniae</italic> was constructed, and after treated with drugs, pathological analysis of lung tissue and determination of bacterial load in lung tissue were performed. The treatment groups included TRQ group, imipenem(IPM) group, TRQ+IPM group, and sterile saline group as the control.</p></sec><sec><title>Results</title><p>The formation of <italic>K. pneumoniae</italic> biofilm was significantly inhibited by TRQ <italic>in vitro</italic> experiments. Furthermore, when combined with IPM, the clearance of <italic>K. pneumoniae</italic> in the biofilm was notably increased compared to the TRQ group and IPM group alone. q-RT PCR analysis revealed that TRQ down-regulated the expression of genes related to biofilm formation in <italic>K. pneumoniae</italic>, specifically <italic>luxS</italic>, <italic>wbbm</italic>, <italic>wzm</italic>, and <italic>lsrK</italic>, and also inhibited the activity of AI-2 molecules in the bacterium. <italic>In vivo</italic> experiments demonstrated that TRQ effectively treated guinea pig lung infections, resulting in reduced lung inflammation. Additionally, when combined with IPM, there was a significant reduction in the bacterial load in lung tissue.</p></sec><sec><title>Conclusion</title><p>TRQ as a potential therapeutic agent plays a great role in the treatment of <italic>K. pneumoniae</italic> infections, particularly in combination with conventional antibiotics. And TRQ can enhanced the clearance effect on the bacterium by inhibiting the <italic>K. pneumoniae</italic> biofilm formation, which provided experimental evidence in support of clinical treatment of TRQ against <italic>K. pneumoniae</italic> infections.</p></sec>