AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite f...AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.展开更多
Objective: To study the therapeutic effect of Com pound Biejia Ruangan prescription (CBRP) on rat model with pulmonary fibrosis in duced by bleomycin. Methods: Fifty four male Sprague Dawley rats were rando mly divide...Objective: To study the therapeutic effect of Com pound Biejia Ruangan prescription (CBRP) on rat model with pulmonary fibrosis in duced by bleomycin. Methods: Fifty four male Sprague Dawley rats were rando mly divided into 6 groups (9 rats in each group). From the first day to the 28th day of the experiment, except to those in the sham model control group that were treated with normal saline, the same amount of bleomycin injection as the n ormal saline given to the control group was given through endotracheal instillat ion to all the rats in all the other groups. From the 29th day of the modeling, CBRP solution of different dosages was respectively injected into the rats in th e high, moderate and low CBRP dose group, while equal volume of normal saline w as given to those in the sham model control group and the model control group , and an equal volume of prednisone solution was given to rats in the prednisone group. On the 80th day, the high resolution computerized tomographic (HRCT) images were observed on an equal footing, and HRCT pathology was correlativel y studied. Results: Different HRCT pathological changes were shown in th e rats with pulmonary fibrosis, such as lung consolidation, thickening of interl obular septum and interlobular mesenchyma as well as lobular deformation, nodule shadow, abnormal brochiovascular tract, thickened pleura with irregular junctio n and polished glass like dense shadows. Honeycomb lung was observed in some cases. Pathological sections showed fibrotic proliferation of lung tissues and noticeable pulmonary interstitial fibrosis. CBRP could improve HRCT images of rats with pulmonary fibrosis, and lower fibrotic p roliferation of the lung tissue.Conclusion: CBRP plays its therapeutic role possibly through its effect on the structure of the lung in rats with pulmonary fibrosis.展开更多
Objective： To investigate the effects of Biejia Ruangan Tablet （复方鳖甲软肝片, BRT）- containing serum on the expression of matrix metalloproteinase （MMP-9） and tissue inhibitor of metalloproteinase （TIMP-1） in...Objective： To investigate the effects of Biejia Ruangan Tablet （复方鳖甲软肝片, BRT）- containing serum on the expression of matrix metalloproteinase （MMP-9） and tissue inhibitor of metalloproteinase （TIMP-1） in cultured renal interstitial fibroblasts. Methods： Different BRT-containing sera were prepared by gastric gavages to rats with the high-dose （7 g/kg）, mid-dose （3.5 g/kg）, and low-dose （1.75 g/kg） BRT respectively. The expression of extracellular matrix in NRK-49F cells was induced by treatment with human transforming growth factor-β1 （recombined human TGF-β 1）, and BRT-containing serum. Western blotting and Northern blotting were used to measure type I and III procollagen, MMP-9, and TIMP-1. Results： The high dose BRT-containing serum could decrease the type Ⅰ and Ⅲ procollagen gene expression which boosted by TGF- 13 1, at the same time cut down TIMP-1 protein and gene expression which increased by TGF- β1 （P〈0.05）. Treatment of cells with recombined human TGF-β 1 had no significant effect on MMP-9 expression and BRT- containing serum also had no effect on MMP-9 expression. Conclusions： High dose BRT has anti-fibrosis effects in NRK-49F cells, as indicated by its inhibition of type Ⅰ and Ⅲ procollagen and TIMP-1 expression.展开更多
Objective:Fufang Biejia Ruangan Tablet(FBRT) is widely used for the treatment of liver fibrosis.However,Hominis Placenta(HP),as an important adjuvant of FBRT,has been restricted for medicinal using due to the limited ...Objective:Fufang Biejia Ruangan Tablet(FBRT) is widely used for the treatment of liver fibrosis.However,Hominis Placenta(HP),as an important adjuvant of FBRT,has been restricted for medicinal using due to the limited availability,ethical controversy and safety issues.The present study aimed to investigate the therapeutic effects of novel FBRT(N-FBRT) with sheep placenta(SP) as substitute for HP on liver fibrosis and explore its possible mechanisms.Different dosages of SP in N-FBRT were also evaluated.Methods:Rats were subcutaneously injected with CCl_(4)to induce liver fibrosis and then treated with NFBRT and FBRT.The anti-hepatic fibrosis effect was determined based on biomarkers analysis of liver function and hepatic fibrosis,and the liver pathology was visualized by H&E staining and Masson staining.The oxidative stress and inflammatory cytokines were also detected.Immunohistochemical staining of a-SMA,real time PCR and Western blotting were performed to evaluate hepatic stellate cells(HSCs)activation and TGF-β1/Smad signaling pathway.Results:N-FBRT and FBRT could ameliorate CCl_(4)-induced liver fibrosis and improve liver function,as evidenced by lowering serum biomarkers levels of liver function and hepatic fibrosis,and decreasing hepatic Hyp content and collagen deposition,and improving the hepatic morphology and architecture changes.Moreover,the anti-liver fibrosis effect was better when the dosage of SP used in N-FBRT was 1/2 of HP in FBRT.Administration of N-FBRT markedly alleviated oxidative stress and inflammatory cytokines,and inhibited a-SMA expression.Furthermore,the mRNA expression of Col Ⅰ,Col Ⅲ,a-SMA and TGF-β1,and proteins expression of a-SMA,TGF-β1,Smad2/3 and p-Smad2/3 were significantly down-regulated by N-FBRT treatment.Conclusion:SP can be used as substitute for HP to prepare N-FBRT for the treatment of liver fibrosis and the anti-liver fibrosis effect of N-FBRT is achieved by eliminating oxidative stress and inflammation,and inhibiting HSCs activation and ECM production by blocking TGF-β1/Smad signaling pathway.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Supported by The Major Project of Applied Basic Research Plan of the Scientific and Technological Department of TianjinChinaNo.06YFJZJC 02900
文摘AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.
基金This study was supported by National Funds of Natur al Science (No.30130220)and Administration of Education against SARS(No.15)
文摘Objective: To study the therapeutic effect of Com pound Biejia Ruangan prescription (CBRP) on rat model with pulmonary fibrosis in duced by bleomycin. Methods: Fifty four male Sprague Dawley rats were rando mly divided into 6 groups (9 rats in each group). From the first day to the 28th day of the experiment, except to those in the sham model control group that were treated with normal saline, the same amount of bleomycin injection as the n ormal saline given to the control group was given through endotracheal instillat ion to all the rats in all the other groups. From the 29th day of the modeling, CBRP solution of different dosages was respectively injected into the rats in th e high, moderate and low CBRP dose group, while equal volume of normal saline w as given to those in the sham model control group and the model control group , and an equal volume of prednisone solution was given to rats in the prednisone group. On the 80th day, the high resolution computerized tomographic (HRCT) images were observed on an equal footing, and HRCT pathology was correlativel y studied. Results: Different HRCT pathological changes were shown in th e rats with pulmonary fibrosis, such as lung consolidation, thickening of interl obular septum and interlobular mesenchyma as well as lobular deformation, nodule shadow, abnormal brochiovascular tract, thickened pleura with irregular junctio n and polished glass like dense shadows. Honeycomb lung was observed in some cases. Pathological sections showed fibrotic proliferation of lung tissues and noticeable pulmonary interstitial fibrosis. CBRP could improve HRCT images of rats with pulmonary fibrosis, and lower fibrotic p roliferation of the lung tissue.Conclusion: CBRP plays its therapeutic role possibly through its effect on the structure of the lung in rats with pulmonary fibrosis.
基金Supported by the National Science Foundation of China(No.30130220 and No.30873345)National Natural Science Found for Innovative Research Groups Science Foundation of China(No.30121005)
文摘Objective： To investigate the effects of Biejia Ruangan Tablet （复方鳖甲软肝片, BRT）- containing serum on the expression of matrix metalloproteinase （MMP-9） and tissue inhibitor of metalloproteinase （TIMP-1） in cultured renal interstitial fibroblasts. Methods： Different BRT-containing sera were prepared by gastric gavages to rats with the high-dose （7 g/kg）, mid-dose （3.5 g/kg）, and low-dose （1.75 g/kg） BRT respectively. The expression of extracellular matrix in NRK-49F cells was induced by treatment with human transforming growth factor-β1 （recombined human TGF-β 1）, and BRT-containing serum. Western blotting and Northern blotting were used to measure type I and III procollagen, MMP-9, and TIMP-1. Results： The high dose BRT-containing serum could decrease the type Ⅰ and Ⅲ procollagen gene expression which boosted by TGF- 13 1, at the same time cut down TIMP-1 protein and gene expression which increased by TGF- β1 （P〈0.05）. Treatment of cells with recombined human TGF-β 1 had no significant effect on MMP-9 expression and BRT- containing serum also had no effect on MMP-9 expression. Conclusions： High dose BRT has anti-fibrosis effects in NRK-49F cells, as indicated by its inhibition of type Ⅰ and Ⅲ procollagen and TIMP-1 expression.
基金financially supported by Inner Mongolia Science and Technology Key Project of China (2015ZY0024)the Chinese Foundation for Hepatitis Prevention and Control Project(WBE20170066)
文摘Objective:Fufang Biejia Ruangan Tablet(FBRT) is widely used for the treatment of liver fibrosis.However,Hominis Placenta(HP),as an important adjuvant of FBRT,has been restricted for medicinal using due to the limited availability,ethical controversy and safety issues.The present study aimed to investigate the therapeutic effects of novel FBRT(N-FBRT) with sheep placenta(SP) as substitute for HP on liver fibrosis and explore its possible mechanisms.Different dosages of SP in N-FBRT were also evaluated.Methods:Rats were subcutaneously injected with CCl_(4)to induce liver fibrosis and then treated with NFBRT and FBRT.The anti-hepatic fibrosis effect was determined based on biomarkers analysis of liver function and hepatic fibrosis,and the liver pathology was visualized by H&E staining and Masson staining.The oxidative stress and inflammatory cytokines were also detected.Immunohistochemical staining of a-SMA,real time PCR and Western blotting were performed to evaluate hepatic stellate cells(HSCs)activation and TGF-β1/Smad signaling pathway.Results:N-FBRT and FBRT could ameliorate CCl_(4)-induced liver fibrosis and improve liver function,as evidenced by lowering serum biomarkers levels of liver function and hepatic fibrosis,and decreasing hepatic Hyp content and collagen deposition,and improving the hepatic morphology and architecture changes.Moreover,the anti-liver fibrosis effect was better when the dosage of SP used in N-FBRT was 1/2 of HP in FBRT.Administration of N-FBRT markedly alleviated oxidative stress and inflammatory cytokines,and inhibited a-SMA expression.Furthermore,the mRNA expression of Col Ⅰ,Col Ⅲ,a-SMA and TGF-β1,and proteins expression of a-SMA,TGF-β1,Smad2/3 and p-Smad2/3 were significantly down-regulated by N-FBRT treatment.Conclusion:SP can be used as substitute for HP to prepare N-FBRT for the treatment of liver fibrosis and the anti-liver fibrosis effect of N-FBRT is achieved by eliminating oxidative stress and inflammation,and inhibiting HSCs activation and ECM production by blocking TGF-β1/Smad signaling pathway.
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.
