RESUMO
The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Assuntos
Humanos , Ciclo Celular , Proliferação de Células , Quinase 1 de Adesão Focal , Genética , Regulação Leucêmica da Expressão Gênica , Células K562 , Ftalazinas , Farmacologia , Piridinas , Farmacologia , RNA Mensageiro , GenéticaRESUMO
<p><b>OBJECTIVE</b>To observe the effect of Shuanghuanglian injection on cerebral expression of nuclear factor-kappaB (NF-kappaB) in mice with viral encephalitis.</p><p><b>METHODS</b>The mice with experimental viral encephalitis received treatment with Shuanghuanglian injection at the dose of 0.2, 1.5, and 5 for 5, 10 or 20 consecutive days. The total RNA of the brain tissue was extracted to analyze the protein and mRNA expression of NF-kappaB using Western blotting and RT-PCR, respectively.</p><p><b>RESULTS</b>Compared with the control group, the mice with experimental viral encephalitis showed significantly increased protein and mRNA expressions of NF-kappaB (P<0.01). Treatment with Shuanghuanglian injection at the doses of 0.2 and 1.5 mg/kg significantly lowered NF-kappaB protein and mRNA expressions in the brain of mice with viral encephalitis (P<0.05), and the effect was even more obvious at the dose of 5 mg/kg (P<0.01).</p><p><b>CONCLUSION</b>Shuanghuanglian injection can reduce the expression of NF-kappaB in the brain of mice with viral encephalitis in a dose- and time-dependent manner.</p>
Assuntos
Animais , Masculino , Camundongos , Western Blotting , Encéfalo , Metabolismo , Medicamentos de Ervas Chinesas , Farmacologia , Usos Terapêuticos , Encefalite Viral , Tratamento Farmacológico , Genética , Metabolismo , Regulação da Expressão Gênica , Injeções , Camundongos Endogâmicos BALB C , NF-kappa B , Genética , Metabolismo , RNA Mensageiro , Genética , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
<p><b>OBJECTIVE</b>To observe the pathological change of partial liquid ventilation (PLV) through establishing the rabbit model of acute lung injury (ALI) induced by meconium aspiration.</p><p><b>METHODS</b>Adult, healthy male or female New Zealand white rabbits were randomly allocated into six groups as follows: (1) control group, (2) conventional mechanical ventilation (CMV) group, (3) high-frequency oscillatory ventilation (HFOV) group, (4) CMV combined with PLV group, (5) HFOV combined with PLV group and (6) normal group. The animals were anesthetized with 1% pentobarbital and tracheotomy was performed and endotracheal tube was placed, 20% meconium fluid (3 ml/kg) was quickly injected into the lung through the endotracheal tube and arterial blood gas was analyzed 30 minutes later. ALI was indicated when P/F ratio (PaO2)/FiO(2)) was < or = 300 mm Hg (1 mm Hg = 0.133 kPa) and Cdyn Dynamic Compliance declined by more than 30% of the baseline. The animals were then randomly allocated into one of the 6 groups. In PLV groups (including CMV + PLV and HFOV + PLV) warmed (37 degrees C) and oxygenated perfluorocarbon was slowly instilled into the lungs of the rabbits through the endotracheal tube at a low-dose 3 ml/kg, then set 15-min positive pressure by sacculus proprius to guarantee perfluorocarbon to steadily diffuse in to the lungs. Six hours after ventilation the animals were sacrificed by using overdose of room air instillation via vein. The lungs were taken and fixed in 4% paraformaldehyde (PFA) and were stained with hematoxylin-eosin (HE). Pathological evaluations included inflammatory manifestation, edema and hemorrhage in both alveolar and interstitial area, damages of small airway (alveolar tube and alveolar bursa) and hyaline membrane formation. One way analysis of variance, Student Newman-Keuls (SNK) method and Kruskal-Wallis (K-W) test were used for comparisons.</p><p><b>RESULTS</b>With the exception of normal group 30 minutes after meconium injections blood gas analysis in different groups showed significant changes and PaO(2)/FiO(2) (< 300 mm Hg), Cdyn declined by more than 60% compared with baseline (P < 0.05). The pathological analysis showed that alveolar and interstitial inflammation, edema, alveolar and interstitial hemorrhage, and small airway damage existed in each group. The hyaline membrane formation was found in one of CMV + PLV group rabbits. The perfluorocarbon-treated animals (CMV + PLV and HFOV + PLV) showed significantly less injury in dependent lung and less damage of small airway (CMV + PLV or HFOV + PLV vs. CMV = 1.1 +/- 0.4 or 0.9 +/- 0.3 vs 2.6 +/- 0.5) compared with the animals of CMV group (P < 0.01). HFOV group (2.1 +/- 0.3) also had less alveolar and interstitial inflammation compared with CMV group (3.0 +/- 0) (P < 0.05), and there was less evidence of alveolar and interstitial edema in the animals treated with HFOV + PLV (1.0 +/- 0.7) compared with CMV (2.0 +/- 0.8) (P < 0.01). Treatment with perfluorocarbon did not result in significant difference in alveolar and interstitial hemorrhage. Compared with CMV and HFOV groups, the groups treated with PLV showed lower mortality of animals (21.4% and 14.3%).</p><p><b>CONCLUSIONS</b>PLV can alleviate the histological damage of acute lung injury induced by meconium aspiration and increased survival chance and therefore PLV would be a useful treatment for MAS. The effectiveness and safety of application of PLV should be evaluated in clinical studies.</p>