RESUMO
OBJECTIVE: To evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin. METHODS: Rats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups. RESULTS: Consistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point. CONCLUSION: Amygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.
Assuntos
Amigdalina/farmacologia , Bleomicina/efeitos adversos , Proteínas Sanguíneas/metabolismo , Fibrose Pulmonar/sangue , Animais , Biomarcadores/sangue , Masculino , Fibrose Pulmonar/induzido quimicamente , Ratos , Ratos WistarRESUMO
OBJECTIVE: To study the effect of Ligustrazine on type I, III collagen synthesis in lung tissue of silica-treated rat. METHODS: The 128 experimental rats were randomly divided into control, silica and Ligustrazine group. 1ml silica (50g/L) was injected intratracheally in silica group and Ligustrazine group, while Ligustrazine group were injected intraperitoneally 50mg/(kg x d) Ligustrazine. Samples were collected on the 1st, 3rd, 7th, 14th, 21st, 28th day after injected silica. Type I , III collagen on paraffin-embedded lung sections were stained with sirius red, detected by polarized light microscopy and quantified by Image-Pro Plus. RESULTS: On the 3rd day after silica instillation, type III collagen began to appear, while type I collagen formed on the 7th day. Thereafter Type I, III collagen increased progressively. Compared with control group, type I, III collagen expression of silica group increased. At different time points type I, III collagen expression of Ligustrazine group decreased than silica group. There was significant difference of type I collagen area percentage on the 7th, 14th, 28th day, type III collagen area percentage on the 28th day between Ligustrazine group and silica group. CONCLUSION: Silica dioxide could induce the increase of type I, III collagen production in lung tissue, and their stages are different. Ligustrazine could suppress type I, III collagen production in lung tissue of silica-treated rat, and it's mechanism would be studied.
Assuntos
Colágeno Tipo III/biossíntese , Colágeno Tipo I/biossíntese , Pulmão/metabolismo , Pirazinas/farmacologia , Dióxido de Silício/toxicidade , Animais , Feminino , Pulmão/efeitos dos fármacos , Masculino , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/prevenção & controle , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
OBJECTIVE: To investigate the change of the expression of the FasL receptor and apoptosis in the pathology of silicosis of the rats exposed to silica and their roles. METHODS: Ninety-six wistar rats were randomizedly divided into the control group and the experimental group. The silicotic animal model was established by the direct tracheal instillation of silica into rat lungs surgically. The control rats underwent directly tracheal instillation of saline into lungs surgically. Eight rats from each group were sacrificed at different days. The expression of FasL receptor in the tissue of the model rats was detected by tissuechip microarray and immunohistochemistry and the cell apoptosis induced by silica was determined by TdT-mediated dUTP nick end-labeling method. The integral optical density of positive cells were quantitatively analyzed using Image-Pro Plus Version 4.5 for windows. RESULTS: The expression of FasL in the lung tissue of the model rats on the 7th, the 14th, the 21st, and the 28th day was significantly higher than that in the control group (P < 0.05), and peaked at the 14th day after exposure to silica. Apoptotic cells in the lung tissue of the model rats on the 1st, the 3rd, the 7th, the 14th, the 21st, and the 28th day were significantly more than those in the control group, and peaked at the 7th and the 14th day after exposure to silica. CONCLUSION: Silica can lead to apoptosis in lung tissues. FasL is expressed in all kinds of cells in the pulmonary tissues of the rats exposed to silica and leads to apoptosis. From the 7th day to 14th day, inflammatory cells dominate in apoptotic cells.
Assuntos
Apoptose , Proteína Ligante Fas/biossíntese , Pulmão/metabolismo , Dióxido de Silício/toxicidade , Silicose/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Pulmão/patologia , Masculino , Distribuição Aleatória , Ratos , Ratos Wistar , Silicose/patologiaRESUMO
OBJECTIVE: To evaluate the effects of taurine in diet on the expression of type I and III collagen and collagen ratio at different time points in rats lung by image process technology. METHODS: Wistar rats were randomly divided into three groups: the saline instilled with a control diet (the saline treated group); silica instilled with a control diet (the silica treated group); and silica instilled with a diet containing 2.5% taurine (the taurine treated group). Animal models were established by the direct tracheal instillation of silica into rat lungs exposed surgically. The taurine concentration of serum was analyzed by means of HPLC. Paraffin embedded lung sections were stained with Sirius red. Polarization microscopy and Image Pro Plus Version 4.5 for windows were used for detecting type I and III collagen. RESULTS: The concentration of taurine in serum of the taurine treated group was significantly elevated compared to the saline treated and silica treated group (P < 0.05 or P < 0.01). Sirius red polarization microscopy showed that type I and III collagen positive area percentage were elevated in the silica treated rats compared with the saline treated group. On the 7th, 14th, 21st, 28th day after silica instillation type I collagen positive area percentage was increased by 3.84, 3.77, 3.73, 9.83 respectively (P < 0.01), and type III collagen positive area percentage were elevated by a little in the silica treated rats compared with saline treated group. The taurine treatment significantly decreased elevation of silica type I collagen positive area percentage of lung by 2.39, 1.62, 7.13 at the 7th, 21st, 28th day respectively (P < 0.05 or P < 0.01), and type III collagen positive area percentage of lung by 2.62 at the 28th day (P < 0.05) compared with the silica treated group. The ratio of type I to III collagen was increased from the 7th day to 28th day after silica instillation, and reached 1.87 at the 28th day with the maximal ratio in the silica-treated group. CONCLUSION: Treatment with taurine can effectively attenuate type I and III collagen expression in the rat lung induced by silica particles at different time points in our study.
Assuntos
Colágeno Tipo III/biossíntese , Colágeno Tipo I/biossíntese , Pulmão/metabolismo , Dióxido de Silício/toxicidade , Taurina/farmacologia , Animais , Feminino , Pulmão/efeitos dos fármacos , Masculino , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
OBJECTIVE: To evaluate the effects of taurine in diet on the expression of inducible nitric oxide synthase (iNOS) in rat lung induced by silica. METHODS: Wistar rats were established by direct tracheal instillation of silica into rat lungs exposed surgically, and the animals of taurine-treated group were silica-instilled with a diet containing taurine. The taurine concentration of serum was analyzed by means of HPLC. The expression of iNOS protein in paraffin-embedded lung sections with Streptavidin/peroxidase (SP) immunohistochemistry on tissue microarray was measured by Image-Pro Plus. RESULTS: The concentration of taurine in serum of taurine-treated group was significantly higher than those in saline-treated and silica-treated groups (P < 0.05). The activities of total NOS and iNOS in BALF supernatant and iNOS positive area percentage of rat lung in silica-treated group were at the peak on 14th day, which were 1.84 U/ml, 1.12 U/ml and 5.42% more respectively than those in saline-treated group (P < 0.05). There were no significant differences between taurine-treated group and silica-treated group in total NOS and iNOS activities of BALF supernatant, and iNOS positive area of the lung (P > 0.05). CONCLUSION: Treatment with taurine hardly influences on the increase in expression of nitric oxide synthase in rat lung induced by silica dust.
Assuntos
Pulmão/enzimologia , Óxido Nítrico Sintase Tipo II/biossíntese , Dióxido de Silício/toxicidade , Taurina/farmacologia , Animais , Líquido da Lavagem Broncoalveolar/química , Feminino , Pulmão/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Taurina/sangueRESUMO
OBJECTIVE: To evaluate the effects of niacin supplemented in diet on temporal expression of nitric oxide synthase in rat lung exposed to silica by tissue array technology. METHODS: Wistar rats were randomly divided into three experimental groups: saline-treated group, silica-treated group, niacin-treated group. There are 48 animals in each group. Animal models were established by direct tracheal instillation of silica into the rat lungs. Plasma level of niacin was measured by high performance liquid chromatography (HPLC). The expression of iNOS protein in the paraffin-embedded lung sections was measured with streptavidin/peroxidase (SP) immunohistochemistry on tissue microarray and quantified by Image-Pro Plus. RESULTS: Plasma level of niacin in niacin-treated group were significantly elevated by 5.946 4, 17.422 0, 21.398 0, 16.091 0, 4.414 3 and 7.130 5 mg/L at 1, 3, 7, 14, 21 and 28 days after instillation of silica, as compared to control and silica-treated groups. Seven days after instillation of silica, iNOS integrated optical density (IOD) of the lung, total NOS and iNOS activities in bronchial alveolar lavage fluid (BALF) supernatant in silica-treated group significantly elevated by 273 421, 2.61 kU/L and 1.89 kU/L, respectively, in the saline-treated group, with statistical significance. Niacin treatment could significantly decrease silica-elevated iNOS integrated optical density (IOD) of the lung, total NOS and iNOS activities in BALF supernatant by 248.292, 1.50 kU/L and 0.91 kU/L in the silica-treated group, respectively, with statistical significance. CONCLUSIONS: It is suggested that treatment with niacin could effectively attenuate the over expression of nitric oxide synthase in the rat lung induced by silica particles in our study.
Assuntos
Niacina/farmacologia , Óxido Nítrico Sintase/metabolismo , Dióxido de Silício/toxicidade , Animais , Líquido da Lavagem Broncoalveolar/química , Feminino , Pulmão/metabolismo , Masculino , Óxido Nítrico Sintase/análise , Óxido Nítrico Sintase Tipo II , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
OBJECTIVE: To evaluate the time-effect of silica on the expression of lung tissue nitric oxide synthase (NOS) in early inflammatory damage stage of silicotic rat. METHODS: Animal models were established by direct tracheal instillation of silica into rat lungs. Total NOS and induced NOS (iNOS) activities in bronchoalveolar lavage fluid (BALF) were assayed. The expression of iNOS protein in paraffin-embedded lung sections with Streptavidin/peroxidase (SP) immunohistochemistry were measured by tissue microarray and Image-Pro Plus. RESULTS: Most of the expression of iNOS was in the cytoplasm of macrophages and neutrophils. iNOS integrated optical density (IOD) of lung tissue increased 1.47 x 10(5) and 2.73 x 10(5) more respectively in silicatreated rats 3, 7 days after exposure than in control rats (P < 0.05), and decreased 1.11 x 10(5) more 28 days after exposure (P < 0.01). The activities of iNOS in BALF increased by 0.86, 1.89 and 0.92 U/ml respectively 3, 7, 14 days after exposure (P < 0.05 or P < 0.01). The activities of total NOS in BALF increased by 1.43, 2.05, 2.61 and 2.19 U/ml respectively 1, 3, 7, 14 days after exposure (P < 0.05 or P < 0.01). CONCLUSION: After silica instillation, the iNOS-positive cells in rat lung tissue were mostly macrophages and neutrophils. There is a parabolic changing trend in the level of expression of lung iNOS during 1 - 28 day exposure to silica.