JaponiconeA induces apoptosis of bortezomib-sensitive and -resistant myeloma cells in vitro and in vivo by targeting IKKβ

Objective:Multiple myeloma(MM)remains incurable with high rates of relapse.New therapeutic drugs are therefore urgently needed to improve the prognosis.JaponiconeA(JA),a natural product isolated from Inula japonica Thunb,has shown good anti-MM potential.A comprehensive study should therefore be conducted to identify both the in vitro and in vivo mechanisms of the anti-MM effects of JA.Methods:CCK8 assays and flow cytometry were used to detect the proliferation,apoptosis,and cell cycle of MM cell lines when treated with JA.In vivo experiments were conducted using subcutaneous xenograft mouse models.We also identified possible targets and the mechanism of JA using RNA-seq and c-Map databases,and identified the specific targets of JA in bortezomib-sensitive and-resistant MM cell lines using CETSA,DARTS,and rescue experiments.Furthermore,JA and bortezomib were used separately or together to characterize their possible synergistic effects.Results:In vitro,JA inhibited proliferation,and induced apoptosis and G2/M phase arrest in MM cell lines,and selectively killed primary CD138+MM cells.In vivo,JA also demonstrated a strong anti-tumor effect with no observable toxicity.In addition,JA showed synergetic effects in combination with bortezomib,and enhanced the anti-tumor effect of bortezomib in bortezomibresistant cells.CETSA and DARTS confirmed direct binding of JA to NF-κB inhibitor kinase beta(IKKβ),and overexpression of IKKβor knockdown of IκBαpartially rescued the apoptosis induced by JA.Conclusions:JA exhibited strong anti-tumor effects in MM.It sensitized myeloma cells to bortezomib and overcame NF-κB-induced drug resistance by inhibiting IKKβ,providing a new treatment strategy for MM patients.

Two new phenolic amides from Allium chinense

Objective: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities.Methods: The chemical constituents were isolated and purified by combining with silica gel column,Sephadex LH-20 column, HPLC and other chromatography techniques. Their structures were elucidated by NMR techniques and mass spectrometry. The isolated compounds were evaluated to determine their protective effect for myocardium cells in vitro.Results: Two new phenolic amides, namely, alichinemide Ⅰ(1) and alichinemide Ⅱ(2), and six konwn amides were isolated from the dried bulbs of A. chinense. The structures of compounds 3–8 were identified as 3-indolcarbaldehyde(3), 1-(2-aminophenyl)urea(4), 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid(5), N-trans-feruloyltyramine(6), N-trans-p-coumaroyltyramine(7), and N-(3,4-dimethoxyphenethyl) acetamide(8). Compound 3(50 μmol/L) showed significant inhibitory effect on the damage of H9c2 myocardial cells induced by H2O2in vitro.Conclusion: Compounds 1 and 2 were new phenolic amides. Compound 3 could be one of the potential myocardium protective constituents of A. chinense.

Camganoids A and B,two new sesquiterpenes with different carbon skeletons isolated from fruits of Cinnamomum migao

Objective:To isolate and identify the undescribed compounds from the fruits of Cinnamomum migao and evaluate its nitric oxide inhibition potential.Methods:The chromatographic techniques of silica gel,Sephadex,and HPLC were used for isolation and purification of the compounds,while HR-ESI-MS,1D NMR,2D NMR,ECD,and X-ray diffraction techniques were used to characterize and confirm the isolated compounds.Moreover,the antiinflammatory activity of the isolated compounds was carried out to check inhibitory potential against the production of nitric oxide with RAW264.7 cells stimulated by LPS.Results:Camganoid A(1),a novel sesquiterpene possessing an unprecedented skeleton,and camganoid B(2),containing a unique eight-membered sesquiterpene moiety with a new carbon skeleton,were isolated and identified from the fruits of C.migao.The absolute configurations of 1 and 2 were confirmed by single crystal X-ray diffraction and electronic circular dichroism(ECD)calculations.Among these compounds,compound 1 exhibited potent inhibitory activity against the production of nitric oxide with IC50value of 4.59μmol/L in RAW264.7 cells stimulated by LPS.Conclusion:The isolation of two new skeletons from the fruits part of C.migao possessed unique skeletons which have not been reported before.

Integrative analysis of microbiome and metabolome in rats with Gest-Aid Plus Oral Liquid supplementation reveals mechanism of its healthcare function

Objective:This study aimed to elucidate the possible mechanism of Gest-Aid Plus Oral Liquid(GAP)on healthcare function.Method:Ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based metabolomics and 16S rDNA sequencing of gut microbiota were performed on serum and fecal samples of GAP and control rats.Additionally,short-chain fatty acids(SCFAs)and inflammatory cytokines in fecal samples were determined through gas chromatography-mass spectrometry and enzyme-linked immunosorbent assay kits.Result:Metabolomics discovered 41 metabolites,which mainly involved amino acid metabolism,lipid metabolism,coenzyme factors,and vitamin metabolism.Administration of GAP increased abundance of Prevotella_9,Alloprevotella,Blautia,Phascolarctobacterium,Parabacteroides,and Fusicatenibacter,and six SCFAs were increased in the GAP group.Measurement of inflammatory cytokines showed that GAP had an anti-inflammatory effect in rats.Conclusions:Administration of GAP greatly affects the aspartate metabolism and microecology of rats,enhances intestinal motility and gut barrier integrity and anti-inflammation.These findings not only have possible implications for further application of GAP,but also provide a link between the gut microbiome,SCFAs,inflammation and serum metabolites in rats.