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Exp Cell Res ; 389(2): 111855, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-31978385


Takeda-G-protein-receptor-5 (TGR5) is a G-protein-coupled receptor (GPCR) activated by bile acids, and mortalin is a multipotent chaperone of the HSP70 family. In the present study, TGR5 was detected by immunohistochemistry (IHC) in extrahepatic cholangiocarcinoma (ECC) specimens, and TGR5 expression in ECC tissues and adjacent tissues was compared. In vitro TGR5 was overexpressed and knocked down in human intrahepatic cholangiocarcinoma (ICC) cell line RBE and human extrahepatic cholangiocarcinoma (ECC) cell line QBC-939 to observe its effects on the biological behavior of cholangiocarcinoma (CC) cells, including proliferation, apoptosis and migration. In vivo xenograft model was constructed to explore the role of TGR5 in CC growth. Proteins that interacted with TGR5 were screened using an immunoprecipitation spectrometry approach, and the identified protein was down-regulated to investigate its contribution to CC growth. The present study demonstrated that TGR5 is highly expressed in CC tissues, and strong TGR5 expression may indicate high malignancy in CC. Furthermore, TGR5 promotes CC cell proliferation, migration, and apoptosis resistance. TGR5 boosts CC growth in vivo. In addition, TGR5 combines with mortalin and regulates mortalin expression in the CC cell line. Mortalin participates in the TGR5-induced increase in CC cell proliferation. In conclusion, TGR5 is of clinical significance based on its implications for the degree of malignancy in patients with CC. Mortalin may be a downstream component regulated by TGR5, and TGR5 promotes cholangiocarcinoma at least partially by interacting with mortalin and upregulating its expression. Both TGR5 and mortalin are positive regulators, and may serve as potential therapeutic targets for CC.

Transl Res ; 182: 88-102, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28034761


Bile acids stimulate intestinal epithelial proliferation in vitro. We sought to investigate the role of the bile acid receptor TGR5 in the protection of intestinal epithelial proliferation in obstructive jaundice. Intestinal tissues and serum samples were obtained from patients with malignant obstructive jaundice and from bile duct ligation (BDL) rats. Intestinal permeability and morphological changes in the intestinal mucosa were observed. The functions of TGR5 in cell proliferation in intestinal epithelial injury were determined by overexpression or knockdown studies in Caco-2 and FHs 74 Int cells pretreated with lipopolysaccharide (LPS). Internal biliary drainage was superior to external biliary drainage in recovering intestinal permeability and mucosal histology in patients with obstructive jaundice. In BDL rats, feeding of chenodeoxycholic acid (CDCA) decreased intestinal mucosa injury. The levels of PCNA, a marker of proliferation, increased in response to CDCA feeding and were paralleled by elevated TGR5 expression. CDCA upregulated TGR5 expression and promoted proliferation in Caco-2 and FHs 74 Int cells pretreated with LPS. Overexpression of TGR5 resulted in increased PCNA, cell viability, EdU incorporation, and the proportion of cells in S phase, whereas knockdown of TGR5 had the opposite effect. Our data indicate that bile acids promote intestinal epithelial cell proliferation and decrease mucosal injury by upregulating TGR5 expression in obstructive jaundice.

Células Epiteliais/metabolismo , Células Epiteliais/patologia , Mucosa Intestinal/lesões , Mucosa Intestinal/metabolismo , Icterícia Obstrutiva/metabolismo , Icterícia Obstrutiva/patologia , Receptores Acoplados a Proteínas-G/metabolismo , Idoso , Animais , Ductos Biliares/efeitos dos fármacos , Ductos Biliares/patologia , Biomarcadores/sangue , Células CACO-2 , Proliferação de Células/efeitos dos fármacos , Ácido Quenodesoxicólico/farmacologia , Células Epiteliais/efeitos dos fármacos , Feminino , Humanos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Icterícia Obstrutiva/sangue , Ligadura , Lipopolissacarídeos , Masculino , Ratos Sprague-Dawley , Regulação para Cima/efeitos dos fármacos
Zhonghua Gan Zang Bing Za Zhi ; 17(9): 653-6, 2009 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-19785950


OBJECTIVE: To investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis. METHODS: Rat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis. RESULTS: (1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively. CONCLUSION: The expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.

Actinas/metabolismo , Cirrose Hepática Biliar/metabolismo , Cirrose Hepática Experimental/metabolismo , Receptores Adrenérgicos alfa/metabolismo , Receptores Adrenérgicos beta/metabolismo , Animais , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Imuno-Histoquímica , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática Biliar/patologia , Cirrose Hepática Experimental/patologia , Masculino , Reação em Cadeia da Polimerase , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptores Adrenérgicos alfa/genética , Receptores Adrenérgicos beta/genética , Sistema Nervoso Simpático/metabolismo , Fatores de Tempo
Zhonghua Yi Xue Za Zhi ; 89(48): 3401-4, 2009 Dec 29.
Artigo em Chinês | MEDLINE | ID: mdl-20223113


OBJECTIVE: To investigate the role of heme oxygenase-1 (HO-1) in the oxidative stress damage of intestinal mucosa barrier disruption in patients with malignant obstructive jaundice (MOJ). METHODS: Fifteen jaundiced patients with malignant biliary obstruction undergoing endoscopic retrograde cholangiopancreatography (ERCP) examination or treatment were enrolled. The control group was comprised of 10 healthy subjects with gastroscopy and 10 patients with non-jaundiced biliary obstruction. Patients were subjected to duodenal biopsy to assess the intestinal oxidative stress as estimated by lipid peroxidation (malondialdehyde) and activity of superoxide dismutase (SOD). Apoptosis of epithelial cell was examined by TdT-mediated dUTP-biotin nick end labeling. Immunohistochemistry and Western blotting were employed to examine the distribution and expression of HO-1 proteins in intestinal mucosa. RESULTS: MOJ jaundiced patients presented high levels of intestinal oxidative stress with a significantly increased level of lipid peroxidation [(1.79 +/- 0.24) vs (1.09 +/- 0.28) vs (1.18 +/- 0.32) nmol x mg(-1) x prot(-1), P = 0.041] and a decreased SOD activity [(303 +/- 10) vs (398 +/- 11) vs (406 +/- 11) nmol x mg(-1) x prot(-1), P = 0.017]. The apoptotic rate of intestinal epithelial cells was significantly higher in jaundiced group than in non-jaundiced control group. Apoptotic index was (69.1 +/- 5.9)%, (28.6 +/- 3.5)% and (10.2 +/- 2.5)% respectively (P < 0.01). The staining of HO-1 was predominantly localized in cytoplasm. In jaundiced patients, HO-1 was obviously elevated than those in the control group (HO-1 optical density 0.28 +/- 0.04 vs 0.20 +/- 0.04 vs 0.13 +/- 0.05) (P < 0.01). Similar outcomes were obtained by quantitative analysis of Western blotting images [HO-1/GAPDH (10.7 +/- 0.7)% vs (7.6 +/- 0.5)% vs (3.9 +/- 0.4)%, P < 0.01]. CONCLUSION: MOJ induces intestinal oxidative stress and it may be a key contributing factor to intestinal barrier failure in the patient population. HO-1 protein level is rising with the progression of obstruction. Perhaps HO-1 has a protective effect upon MOJ through anti-oxidative damage.

Heme Oxigenase-1/metabolismo , Mucosa Intestinal/metabolismo , Icterícia Obstrutiva/metabolismo , Estresse Oxidativo , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose , Estudos de Casos e Controles , Feminino , Humanos , Mucosa Intestinal/patologia , Icterícia Obstrutiva/patologia , Peroxidação de Lipídeos , Masculino , Pessoa de Meia-Idade
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 23(9): 801-3, 2007 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-17825222


AIM: To study the effect of Arg-Gly-Asp-Ser (RGDS) tetrapeptide on the expression of matrix metalloproteinase-13 (MMP-13) and the tissue inhibitor of metalloproteinase-1(TIMP-1) in hepatic stellate cell(HSC) stimulated by fibronectin (FN). METHODS: The expression of MMP-13 was evaluated by RT-PCR. TIMP-1 was analyzed by in-situ hybridization at mRNA level and by Western blot at protein level. RESULTS: Treated by RGDS tetrapeptide for 2 h, the mRNA expression of MMP-13 was upregulated but the expression of TIMP-1 mRNA and its protein in HSC was inhibited. CONCLUSION: The expression of MMP-13 can be induced by RGDS tetrapeptide and the expression of TIMP-1 can be inhibited by RGDS tetrapeptide, which is one of the mechanisms of its anti-fibrogenesis.

Colágeno/metabolismo , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Oligopeptídeos/farmacologia , Animais , Relação Dose-Resposta a Droga , Fibronectinas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Metaloproteinase 13 da Matriz/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo
Artigo em Chinês | MEDLINE | ID: mdl-21180139


AIM: To investigate the effect of IH764-3 on the expression of MMP-13 and TIMP-1 by H2O2-stimulated hepatic stellate cell and the alteration of FAK during this process. METHODS: The expression of MMP-13 and FAK mRNA was examined by RT-PCR. TIMP-1 mRNA was analyzed by in-situ hybridization. FAK and TIMP-1 were evaluated at protein level through Western blotting method. RESULTS: Being incubated for 2 h, compared with control group, MMP-13 mRNA was upregulated by IH764-3, but TIMP-1 transcription was reduced in a dose-dependent manner, accompanied with the decrease of FAK mRNA. The expression of TIMP-1 and FAK protein in HSC also decreased after being exposed by IH764-3 for 24 h. CONCLUSION: IH764-3 can induce the expression of MMP-13 and inhibit the expression of TIMP-1. Down-regulating the expression of FAK mRNA may be one of its mechanisms.

Medicamentos de Ervas Chinesas/farmacologia , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Células Estreladas do Fígado/efeitos dos fármacos , Metaloproteinase 13 da Matriz/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Animais , Células Cultivadas , Regulação para Baixo , Células Estreladas do Fígado/metabolismo , Peróxido de Hidrogênio/efeitos adversos , Ratos , Salvia miltiorrhiza/química