Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephad...Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephadex G-15 gel chromatography and RP-HPLC. One novel anti-hepatic fibrosis peptide （CTEPH-I： Asn-Pro-Asn-Pro-Thr） was obtained and identified. MTS assay and enzyme-linked immunosorbent assay were applied to evaluate the anti-fibrotic effect of CTEPH-1 on activated hepatic stellate cells （HSCs） in vitro. CTEPH-1 efficiently inhibited activation and proliferation of cultured HSC-T6 cells via lowering the contents of collagen and TIMP- 1 except for matrix metalloproteinase- 1 （MMP- 1）. The purified peptide might be beneficial as functional food or potential drug for treatment of liver fibrogenesis.展开更多
This study was aimed to synthesize an active peptide from Carapax Trionycis and to investigate its effect on the proliferation of hepatic stellate cells（HSCs）.An active peptide from Carapax Trionycis,which was shown...This study was aimed to synthesize an active peptide from Carapax Trionycis and to investigate its effect on the proliferation of hepatic stellate cells（HSCs）.An active peptide from Carapax Trionycis,which was shown to have significant anti-hepatic fibrosis activity,was synthesized by solid phase method and characterized by MALDI-TOF MS analysis.The HSCs in log growth phase was treated with the synthetic peptide at different concentrations.Viability and apoptosis of hepatic stellate cells were determined by MTS assay and Annexin V-FITC/PI staining,respectively.The active peptide showed strong inhibition of proliferation and induction of apoptosis of HSC-T6 in a concentration-dependent manner.The results suggest that the active peptide from Carapax Trionycis could be synthesized efficiently and has significant inhibitory effect on the proliferation of HSC-T6.展开更多
OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative b...OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative breast cancer(TNBC)cells based on PI3K/Akt/mTOR signaling pathway.METHODS:MDA-MB-231 cells were treated with different medicated serum as Biejia-,Ezhu-,Biejia-Ezhu(BJ-,EZ-,BJ-EZ-)groups,intervened with no drug rat serum and paclitaxel with final concentration of 33 nM(IC50)as negative and positive control(NC and PC)groups.CCK-8 assay,scratch test,and Transwell assay were used to examine cell proliferation,invasion,and migration.The expression of E-cadherin,N-cadherin,Vimentin,MMP-2,MMP-9,PI3K,Akt,p-Akt,mTOR,and p-mTOR was determined by Western blot,and the m RNA expression of PI3K,Akt and mTOR was determined by real-time polymerase chain reaction.RESULTS:BJ-EZ group inhibited proliferation after 24,48,and 72 h compared with the NC group(P<0.05,<0.01 or<0.001)and reduced the invasion and migration of MDA-MB-231 cells(P<0.01 or<0.001).In addition,BJ-EZ group upregulated the expression of E-cadherin,downregulated the expression of N-cadherin,Vimentin,MMP-2,and MMP-9(P<0.05,P<0.01 or P<0.001),and inhibited the m RNA and protein expression of PI3K,Akt(p-Akt),mTOR(p-mTOR)(P<0.05,<0.01 or<0.001).CONCLUSION:Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine can inhibit the proliferation,invasion,migration and EMT of MDA-MB-231 cells through PI3K/Akt/mTOR signaling pathway,and the effect is better than that of Biejia(Carapax Trionycis)or Ezhu(Rhizoma Curcumae Phaeocaulis)alone.展开更多
基金The Foundation of the Education Department of Hubei Province(Grant No.D20162004)
文摘Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephadex G-15 gel chromatography and RP-HPLC. One novel anti-hepatic fibrosis peptide （CTEPH-I： Asn-Pro-Asn-Pro-Thr） was obtained and identified. MTS assay and enzyme-linked immunosorbent assay were applied to evaluate the anti-fibrotic effect of CTEPH-1 on activated hepatic stellate cells （HSCs） in vitro. CTEPH-1 efficiently inhibited activation and proliferation of cultured HSC-T6 cells via lowering the contents of collagen and TIMP- 1 except for matrix metalloproteinase- 1 （MMP- 1）. The purified peptide might be beneficial as functional food or potential drug for treatment of liver fibrogenesis.
文摘This study was aimed to synthesize an active peptide from Carapax Trionycis and to investigate its effect on the proliferation of hepatic stellate cells（HSCs）.An active peptide from Carapax Trionycis,which was shown to have significant anti-hepatic fibrosis activity,was synthesized by solid phase method and characterized by MALDI-TOF MS analysis.The HSCs in log growth phase was treated with the synthetic peptide at different concentrations.Viability and apoptosis of hepatic stellate cells were determined by MTS assay and Annexin V-FITC/PI staining,respectively.The active peptide showed strong inhibition of proliferation and induction of apoptosis of HSC-T6 in a concentration-dependent manner.The results suggest that the active peptide from Carapax Trionycis could be synthesized efficiently and has significant inhibitory effect on the proliferation of HSC-T6.
基金Supported by the National Natural Science Foundation of China:Based on the"Hu-Chang"Theory,the Mechanism of Shuyu Pill on the Effect of Epithelial-mesenchymal Transition to Inhibit the Metastasis of Triple-negative Breast Cancer by P13K/Akt/mTOR Pathway(No.81960834)。
文摘OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative breast cancer(TNBC)cells based on PI3K/Akt/mTOR signaling pathway.METHODS:MDA-MB-231 cells were treated with different medicated serum as Biejia-,Ezhu-,Biejia-Ezhu(BJ-,EZ-,BJ-EZ-)groups,intervened with no drug rat serum and paclitaxel with final concentration of 33 nM(IC50)as negative and positive control(NC and PC)groups.CCK-8 assay,scratch test,and Transwell assay were used to examine cell proliferation,invasion,and migration.The expression of E-cadherin,N-cadherin,Vimentin,MMP-2,MMP-9,PI3K,Akt,p-Akt,mTOR,and p-mTOR was determined by Western blot,and the m RNA expression of PI3K,Akt and mTOR was determined by real-time polymerase chain reaction.RESULTS:BJ-EZ group inhibited proliferation after 24,48,and 72 h compared with the NC group(P<0.05,<0.01 or<0.001)and reduced the invasion and migration of MDA-MB-231 cells(P<0.01 or<0.001).In addition,BJ-EZ group upregulated the expression of E-cadherin,downregulated the expression of N-cadherin,Vimentin,MMP-2,and MMP-9(P<0.05,P<0.01 or P<0.001),and inhibited the m RNA and protein expression of PI3K,Akt(p-Akt),mTOR(p-mTOR)(P<0.05,<0.01 or<0.001).CONCLUSION:Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine can inhibit the proliferation,invasion,migration and EMT of MDA-MB-231 cells through PI3K/Akt/mTOR signaling pathway,and the effect is better than that of Biejia(Carapax Trionycis)or Ezhu(Rhizoma Curcumae Phaeocaulis)alone.
