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1.
Chinese Journal of Hepatology ; (12): 32-36, 2010.
Artigo em Chinês | WPRIM | ID: wpr-247606

RESUMO

<p><b>OBJECTIVE</b>To explore the effects of HBsAg pulsed dendritic vaccination on anti-HBs production in immunosuppressed rats after liver transplantation (LT).</p><p><b>METHODS</b>Brown-Norway liver allografts were transplanted into Lewis recipients. The transplanted Lewis rats were injected with EK506 (2 mg/kg) and randomly divided into two groups: rats in HBsAg-DCs group (n = 15) were intraperitoneally injected with HBsAg pulsed DCs at 14 d and 28 d after LT, and rats in the HBsAg group (n = 15) were injected with HBsAg (200 mul) once a week for 12 weeks. Rats without any immunosuppressive treatment after LT served as controls (n = 5). IL-2 and IFN-gamma mRNA expression in spleen were analyzed by RT-PCR, serum IL-2, IFN-gamma and anti-HBs were detected by ELISA.</p><p><b>RESULTS</b>High dose of FK506 resulted in the immunosuppressed in LT rats, as evident by low production of IL-2 and IFN-gamma, and without liver rejection compared to rats in the control group. HBsAg-DCs induced high titer of anti-HBs antibody, however, titer of anti-HBs were seldom detectable in the HBsAg group at 1, 2 and 3 mouth after vaccination.</p><p><b>CONCLUSION</b>The capacity of HBsAg-DCs to induce anti-HBs in immunosuppressed rats suggested that DC vaccine may prevent HBV recurrence in liver transplanted patients.</p>


Assuntos
Animais , Masculino , Ratos , Adjuvantes Imunológicos , Farmacologia , Citocinas , Sangue , Genética , Metabolismo , Células Dendríticas , Alergia e Imunologia , Modelos Animais de Doenças , Hepatite B , Alergia e Imunologia , Anticorpos Anti-Hepatite B , Sangue , Alergia e Imunologia , Antígenos de Superfície da Hepatite B , Alergia e Imunologia , Vacinas contra Hepatite B , Terapia de Imunossupressão , Imunossupressores , Transplante de Fígado , Alergia e Imunologia , RNA Mensageiro , Genética , Metabolismo , Distribuição Aleatória , Ratos Endogâmicos BN , Ratos Endogâmicos Lew , Prevenção Secundária , Baço , Alergia e Imunologia , Metabolismo
2.
Chinese Journal of Surgery ; (12): 189-192, 2006.
Artigo em Chinês | WPRIM | ID: wpr-317184

RESUMO

<p><b>OBJECTIVE</b>To explore the possible mechanism and optimal treatment phase of glycine for inhibition lipopolysaccharide (LPS) induced Kupffer cells (KCs) activation.</p><p><b>METHODS</b>The KCs were isolated from 40 BALB/c mice and divided into four groups: the endotoxin group, the prevention group, the early treatment group and the later treatment group (n = 10). The endotoxin group was treated with 10 mg/L LPS, and in the other three groups, glycine (1 mmol/L) was given 24 h before, or at 0 h or 4 h respectively after LPS stimulation. At 0 h, 1 h, 2 h, 6 h and 12 h after LPS stimulation, the mRNA levels and protein expression of interleukin-1 receptor associated kinase-4 (IRAK-4) were determined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively, and nuclear factor-kappaB (NF-kappaB) activities as well as tumor necrosis factor alpha (TNF-alpha) levels were also detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The climax values of IRAK-4, NF-kappaB and TNF-alpha were significantly higher in the endotoxin group and the later treatment group than that in the other two groups (t = 3.17, 4.33, 2.47, 126.73, P < 0.01).</p><p><b>CONCLUSION</b>The results indicated that prophylactic or simultaneous treatment with glycine could effectively inhibit LPS-induced KCs activation by inhibiting IRAK-4 expression.</p>


Assuntos
Animais , Masculino , Camundongos , Células Cultivadas , Interações Medicamentosas , Glicina , Farmacologia , Quinases Associadas a Receptores de Interleucina-1 , Peptídeos e Proteínas de Sinalização Intracelular , Genética , Metabolismo , Células de Kupffer , Metabolismo , Lipopolissacarídeos , Farmacologia , Camundongos Endogâmicos BALB C , NF-kappa B , Metabolismo , Proteínas Serina-Treonina Quinases , Genética , Metabolismo , RNA Mensageiro , Metabolismo , Fatores de Tempo , Fator de Necrose Tumoral alfa , Metabolismo
3.
Chinese Journal of Hepatology ; (12): 574-577, 2006.
Artigo em Chinês | WPRIM | ID: wpr-341297

RESUMO

<p><b>OBJECTIVE</b>To explore the protective mechanisms of glycine (Gly) on lipopolysaccharides (LPS) induced liver injury.</p><p><b>METHODS</b>BABL/c mice were randomly divided into a LPS group, in which the animals were intraperitoneally injected with 10 mg/kg LPS, and a Gly group, in which the mice were pretreated with a 5% Gly-containing diet for 3 days before receiving the same dose of LPS. The livers of the mice were examined for histopathological changes. The TNF alpha and interleukin-10 (IL-10) levels in the blood plasma were measured using ELISA analysis. The mRNA expression of TNF alpha, IL-10 and Toll-like receptor 4 (TLR4) in hepatic tissues were detected using RT-PCR analysis. Protein expression of TLR4 in livers was detected using immunohistochemistry.</p><p><b>RESULTS</b>The Gly group mice had an improved survival rate and attenuated LPS-induced pathological changes in the liver tissues in comparison with those of the LPS group animals. The TNF alpha levels [(1,852.80+/-126.64) pg/ml vs (708.83+/-51.29) pg/ml, P<0.05] in plasma, as well as the expression of TNF alpha (A 1.59+/-0.14 vs. 0.91+/-0.11, P<0.05) and TLR4 (A 0.97+/-0.12 vs. 0.53+/-0.11, P<0.05) mRNA in liver tissues were decreased. However, the levels of plasma interleukin-10 [(344.09+/-31.70) pg/ml vs (418.64+/-38.86) pg/ml, P<0.05] were significantly increased and the peaking time left, shifted.</p><p><b>CONCLUSIONS</b>Gly pretreatment could attenuate LPS -induced liver injury in mice, which may be associated with its role in down-regulating TLR4 expression and up-regulating IL-10 production.</p>


Assuntos
Animais , Feminino , Camundongos , Regulação para Baixo , Glicina , Farmacologia , Interleucina-10 , Sangue , Metabolismo , Lipopolissacarídeos , Fígado , Metabolismo , Patologia , Camundongos Endogâmicos BALB C , Receptor 4 Toll-Like , Metabolismo , Fator de Necrose Tumoral alfa , Sangue , Metabolismo , Regulação para Cima
4.
Chinese Journal of Hepatology ; (12): 97-100, 2006.
Artigo em Chinês | WPRIM | ID: wpr-245740

RESUMO

<p><b>OBJECTIVE</b>To explore the mechanism of endotoxin tolerance (ET) through observing the expression of interleukin 1 receptor associated kinase-4 (IRAK-4) during endotoxin tolerance development in Kupffer cells (KCs).</p><p><b>METHODS</b>Isolated KCs of Balb/c mouse were divided into two groups: the non-endotoxin tolerance (NET) group and the endotoxin tolerance (ET) group, which were pretreated with 10 ng/ml lipopolysaccharide (LPS) for 24 h. Then, the two groups were treated with 100 ng/ml LPS. The expressions of IRAK-4 gene and protein level were determined by RT-PCR and Western blot. The activities of NF-kappaB of KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h after LPS stimulation.</p><p><b>RESULTS</b>The ultimate level of IRAK-4, the activities of NF-kappaB and the TNFalpha level were evidently lower in the ET group than those in the NET group (t = 12.4, 17.4 and 138.9 respectively, P<0.01).</p><p><b>CONCLUSIONS</b>Pretreatment with LPS on KCs could induce endotoxin tolerance of KCs and inhibition of IRAK-4 expression may be one of the reasons for its development.</p>


Assuntos
Animais , Masculino , Camundongos , Células Cultivadas , Endotoxinas , Alergia e Imunologia , Tolerância Imunológica , Quinases Associadas a Receptores de Interleucina-1 , Genética , Células de Kupffer , Biologia Celular , Alergia e Imunologia , Metabolismo , Lipopolissacarídeos , Alergia e Imunologia , Camundongos Endogâmicos BALB C
5.
Chinese Journal of Hepatology ; (12): 819-822, 2005.
Artigo em Chinês | WPRIM | ID: wpr-276342

RESUMO

<p><b>OBJECTIVE</b>To explore the inhibitory effects on the activation of endotoxin-induced Kupffer cells (KCs) through short hairpin RNA (shRNA) targeting interleukin-1 receptor associated kinase-4 (IRAK-4) gene.</p><p><b>METHODS</b>Two effective transfection shRNA plasmid (pSIIRAK-4-A, pSIIRAK-4-B) and one invalidated plasmids (pSIIRAK-4-C) targeting IRAK-4 gene were constructed. The isolated mouse KCs were divided into three groups: the normal control group, the RNAi control group (pSIIRAK-4-C) and the RNAi effective group (pSIIRAK-4-A, pSIIRAK-4-B). Then KCs were stimulated with 0.1 microg/ml lipopolysaccharide (LPS) after 24 h transfection with the constructed plasmid. The expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot at 6 h after LPS stimulation, and the activities of NF-kappaB in KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h.</p><p><b>RESULTS</b>The level of IRAK-4, the activities of NF-kappaB and the TNF-alpha level in the RNAi effective group were evidently lower than those in normal and RNAi control groups (P < 0.01) at 1 h, 3 h, and 6 h. Especially, the pSIIRAK-4-A group in which the changes of the above indices were of no difference (P > 0.05), had better inhibited effects than that of the pSIIRAK-4-B group (P < 0.01).</p><p><b>CONCLUSION</b>The shRNA targeting IRAK-4 gene could effectively inhibit the activation of endotoxin-induced KCs.</p>


Assuntos
Animais , Masculino , Camundongos , Endotoxinas , Quinases Associadas a Receptores de Interleucina-1 , Genética , Metabolismo , Células de Kupffer , Metabolismo , Camundongos Endogâmicos BALB C , Interferência de RNA , RNA Interferente Pequeno , Genética , Transdução de Sinais , Fisiologia
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