RESUMO
OBJECTIVE: To observe the protective effects of safflor Injection (SI) and extract of Ginkgo biloba (EGB) on lung ischemia-reperfusion injury (LIRI) and investigate its mechanism. METHODS: In vivo rabbit model of LIRI was reconstructed. Forty rabbits were randomly and equally divided into four groups: sham-operation group (sham group), ischemia-reperfusion group (model group), ischemia-reperfusion plus SI group (safflor group) and ischemia-reperfusion plus EGB injection group (EGB group). Malondialdehyde (MDA) content, superoxide dismutase (SOD) and xanthine oxidase (XO) activity in serum were measured. The wet/dry weight ratio (W/D) of the lung tissue and activity of myeloperoxidase (MPO) were also tested. Ultrastructure change of the lung tissue was observed by the electron microscope. The expression of intercellular adhesion molecule-1 (ICAM-1) was measured by immunohistochemistry (IHC). RESULTS: In the model group, MDA and XO increased and SOD decreased in serum compared with the sham group (P<0.01). The values of W/D, MPO and ICAM-1 of the model group were higher than those of the sham group (P<0.01), but those of the safflor group and EGB group were significantly lower than those of the model group (P<0.01). The IHC demonstrated that ICAM-1 expression in lung tissue of the model group was significantly higher than those of the safflor group (P<0.01). Compared with safflor group, in the EGB group MDA, XO, MPO decreased, SOD and ICAM-1 expression increased (P<0.05), but the change of W/D was not statistically significant (P>0.05). CONCLUSIONS: SI and EGB may attenuate LIRI through antioxidation, inhibition of neutrophil aggregation and down-regulation of ICAM-1 expression. But EGB had more effect on the antioxidation, while SI did better on regulating ICAM-1 expression.
Assuntos
Ginkgo biloba/química , Pulmão/irrigação sanguínea , Extratos Vegetais/uso terapêutico , Substâncias Protetoras/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Óleo de Cártamo/uso terapêutico , Animais , Feminino , Imuno-Histoquímica , Injeções , Molécula 1 de Adesão Intercelular/metabolismo , Pulmão/patologia , Masculino , Malondialdeído/metabolismo , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/farmacologia , Coelhos , Traumatismo por Reperfusão/sangue , Óleo de Cártamo/administração & dosagem , Óleo de Cártamo/farmacologia , Superóxido Dismutase/sangue , Xantina Oxidase/sangueRESUMO
AIM: Excessive production of inflammatory mediators during invasive infection plays a key role in the pathogenesis of sepsis. In an attempt to improve survival of patients with this lethal syndrome, agents were developed to selectively inhibit mediators in this inflammatory response. Ulinastatin (UTI), a human protease inhibitor, inhibits the enhanced production of pro-inflammatory molecules. However, it is unknown if Ulinastatin treatment could result in protective effects for sepsis. The aim of this study was to investigate the role of Ulinastatin on septic rats. METHODS: Sixty male Wistar rats were divided into six groups, 10 of each: sham-operation plus PBS (5 ml), cecal ligation and puncture (CLP) plus PBS (5 ml), CLP plus UTI (5000 U/kg), CLP plus UTI (10,000 U/kg), CLP plus UTI (20,000 U/kg) and sham-operation plus UTI (10,000 U/kg). Rats in the UTI groups after CLP operation were treated with Ulinastatin by intraperitoneal injection at different doses and then compared with untreated sepsis control animals. RESULTS: The intestinal concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-13 (IL-13) were significantly higher in septic rats than those in normal rats. Ulinastatin administration effectively suppressed the levels of TNF-α and IL-6, whereas it markedly enhanced the levels of IL-10 and IL-13. CONCLUSION: Ulinastatin may possess a protective role in the septic process by inhibiting TNF-α and IL-6, and augmenting IL-10 and IL-13 concentrations in intestine of septic rats.
Assuntos
Anti-Inflamatórios/farmacologia , Glicoproteínas/farmacologia , Sepse/prevenção & controle , Inibidores da Tripsina/farmacologia , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Interleucina-1/biossíntese , Interleucina-10/biossíntese , Interleucina-6/biossíntese , Masculino , Ratos , Ratos Wistar , Sepse/imunologia , Fator de Necrose Tumoral alfa/biossínteseRESUMO
OBJECTIVE: To construct three recombinant shuttle plasmids of adenovirus expression vector which can express hepatitis C virus(HCV) different structure genes(C, C+E1, C+E1+E2) in order to pack adenovirus expression vectors which can express HCV different structure gene effectively. METHODS: The different HCV structure genes derived from the plasmid pBRTM/HCV1-3011 by using polymerase chain reaction (PCR) were inserted into the backward position of cytomegalovirus(CMV) immediate early promotor element of shuttle plasmid(pAd.CMV-Link.1) of adenovirus expression vector respectively, then the three recombinant plasmids (pAd.HCV-C, pAd.HCV-CE1, pAd.HCV-S) were obtained. The recombinant plasmids were identified by endonuclease, PCR and sequencing. HCV structure genes were expressed transiently with Lipofectamine 2000 coated in HepG2 cells which were confirmed by immunofluorescence and Western-Blot. RESULTS: Insert DNAs of the three recombinant plasmids' were confirmed to be HCV different structure genes by endonuclease, PCR and sequencing. The three recombinant plasmids can express HCV structure gene (C, C+E1, C+E1+E2) transiently in HepG2 cells which were confirmed by immunofluorescence and Western-Blot. CONCLUSION: The three recombinant shuttle plasmids of adenovirus expression vector can express HCV structure gene(C, C+E1, C+E1+E2) transiently. This should be useful to pack adenovirus expression vector which can express HCV structure genes.
Assuntos
Adenoviridae/genética , Vetores Genéticos , Hepacivirus/genética , Proteínas Estruturais Virais/genética , Expressão Gênica , Estrutura Molecular , PlasmídeosRESUMO
AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P<0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.
Assuntos
Imunoensaio/métodos , Cirrose Hepática/diagnóstico , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Anticorpos Monoclonais/metabolismo , Eritrócitos/metabolismo , Humanos , Hibridização In Situ , Fígado/metabolismo , Cirrose Hepática/metabolismo , Inibidores de Proteases/metabolismo , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-2/genéticaRESUMO
Objective. To investigate the rule of intensity and duration of HSP70 expression in rat brain and its relationship with brain injury after repeated +Gz exposures. Method. SD male rats were arranged into control group, +2 Gz, +4 Gz, +6 Gz, and +10 Gz exposure groups. Rat brains were taken 6 h, 10 h, 1 d, 2 d, 4 d or 6 d after +Gz exposure for histopathologic and immunohistochemic or in situ hybridization studies. The expression of HSP70 and HSP70 mRNA or morphology of neurons were observed. Result. The intensity and duration of HSP70 expression were weak and brief at +2 Gz exposure, but was relatively extensive. There was a middling reaction of HSP70 only in hippocampal area after +10 Gz exposure. The duration, extension and intensity of HSP70 expression were wide, long and strong after +4 Gz and +6 Gz exposures. After 1 or 3-5 times exposures, the HSP70 expression reached its peak on the first day after +4 Gz exposures, and dropped obviously on the second day. However the expression of HSP70 maintained a high level after 2 d and was still higher than normal on the 6 d after 3-5 times repeated +4 Gz exposures. The distribution of HSP70 mRNA expression was as same as that of the HSP70 expression but the peak appeared much earlier (10 h) and its duration was shorter. After +10 Gz/5 min exposure, degenerated neurons were found in cortex, hippocampus and thalamus regions while the number of degenerated neurons were obviously decreased in such areas in pre-exposure groups with repeated +4 Gz/3 min for 3-5 times. Conclusion. The intensity and duration of HSP70 and HSP70 mRNA expression after +4 Gz and +6 Gz exposure were stronger and longer than +2 Gz and +10 Gz exposure. The degree of neuron damage after +10 Gz/5 min exposure in pre-exposure groups with repeated +4 Gz/3 min 3-5 times was obviously slight comparing with that of single +10 Gz exposure group.