Efficacy of Tounongsan decoction(透脓散方)on pyogenic liver abscess:network pharmacology and clinical trial validation

OBJECTIVE:To elucidate the molecular mechanisms governing the effect of Tounongsan decoction(透脓散方,TNS)on the pyogenic liver abscess.METHODS:Based on oral bioavailability and druglikeness,the main active components of TNS were screened using the Traditional Chinese Medicine Systems Pharmacology platform.The Gene Card and Uni Prot databases were used to establish a database of pyogenic liver abscess targets.The interactive network map of drug-ingredients-target-disease was constructed using Cytoscape software(Version 3.7.2).A proteinprotein interaction network was constructed using the STRING database,and the related protein interaction relationships were analyzed.biological process of gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed for the core targets.Finally,a clinical trial was performed to verify the reliability of the network pharmacology.RESULTS:Forty active components of TNS decoction were obtained,and 61 potential targets and 11 proteins were identified.Pathways involved in the treatment of pyogenic liver abscess include the phosphatidylinositide 3-kinases-protein kinase B(PI3K-AKT),advanced glycation end products-receptor for advanced glycation end products(AGE-RAGE),and tumor necrosis factor(TNF)signaling pathways.The results of network pharmacology analysis combined with clinical trials validated that TNS decoction could alleviate the inflammatory response of pyogenic liver abscesses by decreasing interleukin 6(IL-6)levels.CONCLUSIONS:TNS decoction has the characteristics of being multi-system,multi-component,and multi-target.Active ingredients in TNS,such as quercetin,kaempferol,fisetin,andβ-sitosterol,have strong potential to be candidate drugs for treating pyogenic liver abscesses.The possible mechanism of TSN decoction includes regulating immune and inflammatory responses and reducing IL-6 production to control inflammatory development.

Tounongsan (透脓散) Extract Induces Apoptosis in Cultured Raji Cells

Objective： To investigate the effects of Tounongsan （透脓散） extract （TNSE） on proliferation and apoptosis of the human lymphoma cell line Raji and its possible mechanism of action. Methods： The viability of TNSE-treated Raji cells was measured by a 3-（4,5-dimethyl-thiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was determined by flow cytometry. The molecular mechanisms of TNSE-mediated apoptosis were further investigated by reverse transcription-polymerase chain reaction （RT-PCR） analysis of the mRNA expression of nuclear factor K B （NF-κB）, Bcl-XL, Bcl-2-associated death promoter （Bad）, caspase-9 and caspase-3. Western blotting was used to detect the protein expressions of NF- κB, Bad, cleaved caspase-9 and cleaved caspase-3. Results： TNSE inhibited Raji cell proliferation in dose- and time-dependent manners. After 48-h treatment with various concentrations of TNSE （125, 250 and 500 μg/mL）, the apoptosis rates of Raji cell were 12.23% ±1.98% （P〈0.05）, 20.97% ± 3.96% （P〈0.01） and 30.4% ± 4.87% （P〈0.01）, respectively, compared with those of the control （6.02% ± 1.01%）. RT-PCR demonstrated that NF- κ B mRNA expression was significantly downregulated in Raji cells treated with 250 μg/mL TNSE for 48 h （P〈0.06）, while Bad, caspase-9 and caspase-3 mRNA levels were upregulated （P〈0.05）. Moreover, TNSE treatment resulted in downregulation of NF- κ B protein expression and strikingly upregulated protein expressions of Bad, cleaved caspase-9, cleaved caspase-3 in a dose-dependent manner, as determined by Western blot. Conclusion： TNSE exhibits significant anti-proliferative and apoptotic effects in Raji cells, which may be involved in regulation of NF- κB and Bad, and activation of caspase-9 and caspase-3.