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Network pharmacology and liver fibrosis(LF) model in vitro were used to analyze the underly mechanism of anti-liver fibrosis effect that induced by Piperis Longi Fructus and its major active compounds. TCMSP and TCMIP were used to search for the chemical constituents of Piperis Longi Fructus, as well as the oral bioavailability(OB), drug-likeness(DL), intercellular permeability of intestinal epithelial cells(Caco-2) and Drug-likeness grading were set as limiting conditions. The related target genes of Piperis Longi Fructus were queried by TCMSP database, while related targets of LF were screened by GeneCards databases. Interaction network was constructed using Cytoscape 3.7.1. These above data were imported into STRING database for PPI network analysis. Enrichment of gene ontology(GO) and pathway analysis(KEGG) within Bioconductor database were utilized to note functions of related targets of Piperis Longi Fructus. Finally, the core targets and pathways were preliminarily verified by in vitro experiments. The effects of piperlongumine(PL), the major active component of Piperis Longi Fructus, on proliferation of rat liver stellate cells(HSC-T6) and expression of α smooth muscle actin(α-SMA) and collagen Ⅰ were investigated. The major factors TNF-α of tumor necrosis factor(TNF) pathway and NF-κB p65, IL-6 protein expressions of LF process were examined. A total of 12 active compounds such as PL were obtained by analyzing the bioavailability and drug-like properties, which inferred to 48 targets. The functional enrichment analysis of GO obtained 1 240 GO items, mainly involving in process of biology and molecular function. A total of 99 signaling pathways were enriched in the KEGG pathway enrichment analysis, including TNF signaling pathway, cGMP-PKG signaling pathway, calcium signaling pathways. CCK-8 assay showed that PL inhibited proliferation of HSC-T6 induced by transforming growth factor-β1(TGF-β1). Western blot analysis found that treated with PL suppressed the protein expressions of α-SMA, collagen Ⅰ, TNF-α and p65 in HSC-T6. Enzyme linked immunosorbent assay(ELISA) showed that PL inhibited the expressions of TNF-α and IL-6 in the cluture supertant of HSC-T6 cells. In conclusion, PL could play an anti-liver fibrosis role by regulating TNF/NF-κB signaling pathway. This study provided the mechanism basis of anti-LF effects induced by Piperis Longi Fructus and its major active compounds, which might help for the further study of the mechanism and key targets of Piperis Longi Fructus.
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Animales , Humanos , Ratas , Células CACO-2 , Células Estrelladas Hepáticas/metabolismo , Cirrosis Hepática/genética , FN-kappa B/metabolismo , Transducción de SeñalRESUMEN
Objective: To investigate whether piperlongumine can sensitize prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trigger apoptosis in prostate cells. Methods: Human prostate cancer cell lines PC3, LNCaP, and VCaP were cultured with piperlongumine and TRAIL. Then, cell proliferation, migration, caspase activation, apoptotic protein expressions, and death receptor expressions were measured. Results: Piperlongumine inhibited cell proliferation at low doses (<10 μM) alone and in combination with TRAIL (25 ng/mL), induced apoptosis, and suppressed cyclooxygenase activation. Additionally, piperlongumine induced expression of death receptors which potentiated TRAIL-induced apoptosis in cancer cells but did not affect decoy receptors. Piperlongumine also downregulated tumor cell-survival pathways, inhibited colony formation and migration of cancer cells alone or in combination with TRAIL. The combination of piperlongumine with TRAIL was found to be synergistic. Conclusions: Our findings indicate that piperlongumine can sensitize cancer cells to TRAIL through the upregulation of death receptors and can trigger apoptosis with the downregulation of anti-apoptotic proteins.
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Background: Piperlongumine (PL) is an alkaloid compound extracted from piperlongum. Many studies have shown that PL has anti-tumor effects on a variety of tumor cells in vivo and in vitro. However, the specific mechanism needs to be further explored. Aims: To investigate the regulatory mechanism of PL on the expression of TERT in gastric cancer cells. Methods: Gastric cancer cells were treated with different doses of PL, AG490, respectively. CCK-8 and plate colony formation experiment were used to detect cell viability. Real-time fluorescent quantitative PCR was used to detect the expression of TERT mRNA. Western blotting was used to detect the protein expressions of TERT, STAT3, p-STAT3 and DNMT1. TRAP-ELISA was used to determine the telomerase activity. Luciferase reporter gene was used to detect the TERT promoter activity. Results: Compared with control group, gastric cancer cells viability in PL group was significantly decreased, colony formation ability was significantly reduced, TERT mRNA and protein expressions, as well as telomerase activity were significantly reduced, p-STAT3 and DNMT1 protein expressions were significantly downregulated. AG490 significantly inhibited gastric cancer cells viability, protein expressions of p-STAT3, DNMT1 and TERT. Conclusions: PL may inhibit gastric cancer cells viability through regulation of TERT expression via STAT3-mediated epigenetic regulation and it may become a new target drug for the treatment of gastric cancer in future.
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The piperlongumine is a tertiary amine amide alkaloid, which is mainly found in the roots of the Piper family like long piper (Piper longum L.) and tumor-like piper (Piper tuberculatum Jacq). It has a variety of pharmacological activities, such as anti-platelet aggregation, anti-inflammatory and anti-tumor and its derivatives also have a variety of activities. In this paper, the research progress on the structural modification and bioactivity of pharmacological effects of the piperlongumine amide is reviewed, which provides a reference for the further study of amides.
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OBJECTIVE: To investigate the inhibitory effect of different concentration of piperlongumine on the growth of lung cancer cells, and further study the effect of piperlongumine on the apoptosis and pyroptosis of NCI-H460 cancer cells by increasing ROS, and elucidate its new anti-tumor mechanism. METHODS: NCI-H460 lung cancer cells were cultured by conventional method, the control group and different concentration of piperlongumine treatment group were set up. MTT assay was used to evaluate the activity of NCI-H460 lung cancer cells, and cell colony assay was performed for the effect of colony formation of NCI-H460 cells. The NCI-H460 cells apoptosis and the growth inhibition of NCI-H460 cells via promotion of ROS levels were evaluated by flow cytometry, respectively. Western blotting was used to detect the expression of Bax, Bcl-2 and caspase-3 proteins after treated with piperlongumine. The effect of piperlongumine on the ROS-based NCI-H460 lung cells pyroptosis was confirmed by microscopic imaging. At the same time, the effect of ROS on apoptosis-related protein GSDME was detected by Western blotting. RESULTS: Piperlongumine inhibited lung cancer cells in a concentration dependent manner, with the IC50 value on NCI-H460 lung cancer of 13.1 μmol.L-1. Piperlongumine restrained the NCI-H460 colony formation in a concentration-dependent pattern and promoted NCI-H460 apoptosis. Piperlongumine promoted the expression of the pro-apoptotic proteins Bax and Cle-caspase-3, and inhibited the expression of the anti-apoptotic protein Bcl-2 in a concentration-dependent manner. Piperlongumine induced NCI-H460 cells apoptosis and pyroptosis by increasing intracellular ROS content, and ROS increased GSDME-N protein expression. CONCLUSION: Piperlongumine inhibits the growth of lung cancer cells, and induces apoptosis and pyroptosis of NCI-H460 cancer cells by promoting the levels of intracellular ROS.
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Objective: To investigate whether piperlongumine can sensitize prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trigger apoptosis in prostate cells.Methods: Human prostate cancer cell lines PC3, LNCaP, and VCaP were cultured with piperlongumine and TRAIL. Then, cell proliferation, migration, caspase activation, apoptotic protein expressions, and death receptor expressions were measured. Results: Piperlongumine inhibited cell proliferation at low doses (<10 μM) alone and in combination with TRAIL (25 ng/mL), induced apoptosis, and suppressed cyclooxygenase activation. Additionally, piperlongumine induced expression of death receptors which potentiated TRAIL-induced apoptosis in cancer cells but did not affect decoy receptors. Piperlongumine also downregulated tumor cell-survival pathways, inhibited colony formation and migration of cancer cells alone or in combination with TRAIL. The combination of piperlongumine with TRAIL was found to be synergistic. Conclusions: Our findings indicate that piperlongumine can sensitize cancer cells to TRAIL through the upregulation of death receptors and can trigger apoptosis with the downregulation of anti-apoptotic proteins.
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Objective To observe the inhibition of piperlongumine in vitro on platelet aggregation and blood coagulation tests for children,to provide the experimental basis for clinical medication.Methods Venous blood samples from 30 children were randomly devided into 5 groups,and was centrifuge to separate platelet-rich plasma (PRP).After storing in 37 ℃ thermostat water bath for 5 minutes,the PRP which have been added DMSO as blank group,and added Aspirin (10 μmol/L)as control group,and added PL (20 μmol/L),PL(100 μmol/L),PL(200 μmol/L) as different concentrations of PL groups respectively,were induced by the addition of adenosine diphosphate (10 μmol/L),collagen(2.5 μg/mL) and the arachidonic acid(500 μg/mL).Then the platelet aggregation rate of the PRP from 5 groups could be measured by turbidimetry.Blood plasma isolated from venous blood was divided into 5 groups.In the PL groups,blood plasma were mixed up with PRP concentrations of which were 5,10,20 μmol/L.In the bland group,blood plasma were mixed up with DMSO (1％).In the control group,blood plasma were mixed up with heparin sodium(35 U/mL).After storing in 37 ℃ thermostat water bath for 5 minutes,fibrinogen(FIB),prothrombin time(PT),thrombin time(TT) and activated partial thromboplastin time of different groups were detected.Results Compared to the control group,the groups which were add PL with different concentrations (20,100,200 μmol/L) showed significant inhibition on platelet aggregation induced by AA and collagen(P<0.05).PL with concentrations of 100 μmol/L and 200 μmol/L showed significant inhibition on platelet aggregation induced by ADP(P<0.05).The PT,APTT,TT of blood plasma from children had been significantly prolonged by the intervention of PL 10 μmol/L and PL 20 μmol/L(P<0.05),however,no significant change of FIB was observed.Conclusion There are inhibitory effects of PL on platelet aggregation of blood plasma from children and anticoagulant activity in this study.
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Background:Recently,studies have shown that piperlongumine( PL)selectively killed cancer cells by elevating reactive oxygen species(ROS)in various cancers. However,the effect of PL on gastric cancer cells remained to be further studied. Aims:To investigate the effect of PL on proliferation and apoptosis of human gastric cancer cell line MKN45 and its underlying mechanism. Methods:MKN45 cells were treated with different doses of PL,caspase inhibitor,antioxidant, and their combinations,respectively. Cell viability was assessed by CCK-8 assay;cell cycle,apoptosis and intracellular ROS level were measured by flow cytometry;and Western blotting was employed to determine the expression of apoptosis-related proteins( XIAP,cleaved-caspase3,7,9 and cleaved-PARP),p53 and its downstream target genes( p21, GADD45α and PUMA). Results:PL inhibited the proliferation of MKN45 cells in a dose- and time-dependent manner. In MKN45 cells treated with PL,the proportion of cells in G1 phase,apoptotic rate and intracellular ROS level were significantly increased,the expression of inhibitor of apoptosis protein XIAP was down-regulated,and the caspase-dependent apoptosis pathway,p53 and its downstream target genes were activated. Pretreatment with antioxidant NAC or Z-VAD-FMK, a general caspase inhibitor could partially abolish the effect of PL on ROS production and its antitumor effect. Conclusions:PL can inhibit cell proliferation and induce cell cycle G1 phase arrest and apoptosis in MKN45 cells. Its antitumor effect may be associated with a ROS-mediated p53 activation and subsequent triggering of caspases cascade of cell apoptosis.
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Piperine and piperlongumine, alkaloids having diverse biological activities, commonly occur in roots of Piper longum L., Piperaceae, which have high commercial, economical and medicinal value. In present study, rapid, validated HPTLC method has been established for the determination of piperine and piperlongumine in methanolic root extract and its commercial formulation 'Mahasudarshan churna®' using ICH guidelines. The use of Accelerated Solvent Extraction (ASE) as an alternative to conventional techniques has been explored. The methanol extracts of root, its formulation and both standard solutions were applied on silica gel F254 HPTLC plates. The plates were developed in Twin chamber using mobile phase toluene: ethyl acetate (6:4, v/v) and scanned at 342 and 325 nm (λmax of piperine and piperlongumine, respectively) using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of piperine and piperlongumine in the range of 20-100 and 30-150 ng/spot, respectively; the correlation coefficient was 0.9957 and 0.9941 respectively. Sharp, symmetrical and well resolved peaks of piperine and piperlongumine spots resolved at Rf 0.51 and 0.74, respectively from other components of the sample extracts. The HPTLC method showed good linearity, recovery and high precision of both markers. Extraction of plant using ASE and rapid HPTLC method provides a new and powerful approach to estimate piperine and piperlongumine as phytomarkers in the extract as well as its commercial formulations for routine quality control.