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<p><b>OBJECTIVE</b>To investigate the effect of panax notoginseng saponins (PNS) injection on pulmonary artery pressure and the expression of p38MAPK in lung tissue of rats subjected to chronic hypoxia.</p><p><b>METHODS</b>Thirty adult male Sprague Dawley rats were randomly divided into three groups (ten in each group): rats in control group were exposed to normoxic condition and the rats in hypoxia group and PNS group were subjected to 4-week hypoxia, and PNS injection (50 mg · kg(-1) · d(-1)) was administrated intraperitoneally at 30 min in the PNS group daily before the rats were kept in the hypoxic chamber, while rats in the other two groups received equal dose of normal saline instead. After chronic hypoxia, mean pulmonary artery pressure (mPAP) and mean carotid artery pressure (mCAP) were measured. The heart and lung tissues were harvested, and right ventricle (RV) and left ventricle plus ventricular septum (LV+S) were weighed to calculate the ratio of RV/(LV+S). The expression of p38MAPK mRNA was determined by reverse transcription-polymerase chain reaction, the quantity of phosphorylated p38MAPK (p-p38MAPK) in rat lung tissues and pulmonary arterioles was determined by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>Compared with the control group, mPAP and the ratio of RV/(LV+S) in the hypoxia group were increased, the expression of p-p38MAPK in pulmonary arterioles and p38MAPK mRNA in the lung were higher (P<0.05). The changes of these parameters in the hypoxia group were significantly attenuated by PNS treatment (P<0.05).</p><p><b>CONCLUSION</b>PNS injection was shown to prevent hypoxic pulmonary hypertension at least partly by regulating p38MAPK pathway.</p>
Assuntos
Animais , Masculino , Arteríolas , Metabolismo , Pressão Sanguínea , Western Blotting , Artérias Carótidas , Modelos Animais de Doenças , Ventrículos do Coração , Hemodinâmica , Hipertensão Pulmonar , Hipóxia , Injeções , Pulmão , Patologia , Sistema de Sinalização das MAP Quinases , Panax notoginseng , Química , Artéria Pulmonar , RNA Mensageiro , Genética , Metabolismo , Ratos Sprague-Dawley , Saponinas , Farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno , Genética , MetabolismoRESUMO
The aim of the present study was to investigate the roles of calcium-activated chloride channels (Cl(Ca)) in the two-phase hypoxic pulmonary vasoconstriction (HPV). The second pulmonary artery branches were dissected from male Sprague-Dawley rats, and the changes in vascular tone were measured by using routine blood vascular perfusion in vitro. The result showed that, under normoxic conditions, Cl(Ca) inhibitors (NFA and IAA-94) significantly relaxed second pulmonary artery contracted by norepinephrine (P < 0.01), but merely had effects on KCl-induced second pulmonary artery contractions. A biphasic contraction response was induced in second pulmonary artery ring pre-contracted with norepinephrine exposed to hypoxic conditions for at least one hour, but no biphasic contraction was observed in pulmonary rings pre-contracted with KCl. NFA and IAA-94 significantly attenuated phase II sustained hypoxic contraction (P < 0.01), and also attenuated phase I vasodilation, but had little effect on phase I contraction. These results suggest that Cl(Ca) is an important component forming phase II contraction in secondary pulmonary artery, but not involved in phase I contraction.
Assuntos
Animais , Masculino , Ratos , Canais de Cloreto , Fisiologia , Glicolatos , Farmacologia , Hipóxia , Norepinefrina , Farmacologia , Artéria Pulmonar , Ratos Sprague-Dawley , Vasoconstrição , VasodilataçãoRESUMO
<p><b>OBJECTIVE</b>To investigate the role of p38 MAPK on ischemic postconditioning (IPO) attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI).</p><p><b>METHODS</b>Forty adult male SD rats were randomly divided into 5 groups based upon the intervention (n = 8): control group (C), LIR group (I/R), LIR + IPO group (IPO), IPO + solution control group (D), IPO + SB203580 group (SB). Left lung tissue was isolated after the 2 hours of reperfusion, the ratio of wet lung weight to dry lung weight (W/D), and total lung water content (TLW) were measured. The histological structure of the left lung was observed under light and electron transmission microscopes, and scored by alveolar damage index of quantitative assessment (IQA). Apoptosis index (AI) of lung tissue was determined by terminal deoxynuleotidyl transferase mediated dUTP nick end and labeling (TUNEL) method. The mRNA expression and protein levels of and Bax were measured by RT-PCR and quantitative immunohistochemistry (IHC).</p><p><b>RESULTS</b>Compared with C group, W/D, TLW, IQA, AI and the expression of Bax of I/R were significantly increased, the expression of Bcl-2 and Bcl-2/Bax were significantly decreased (P < 0.05, P < 0.01), and was obviously morphological abnormality in lung tissue. Compared with I/R group, all the indexes of IPO except for the expression of Bcl-2 and Bcl-2/ Bax were obviously reduced, the expression of Bcl-2 and Bcl-2/Bax were increased (P < 0.05, P < 0.01). All the indexes between D and IPO were little or not significant( P > 0.05). The expression of Bcl-2 and Bcl-2/Bax of SB were significantly increased and other indexes were reduced than those of IPO (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>IPO may attenuate pneumocyte apoptosis in LIRI by inactivation of p38 MAPK, up-regulating expression of Bcl-2/Bax ratio.</p>
Assuntos
Animais , Masculino , Ratos , Células Epiteliais Alveolares , Biologia Celular , Apoptose , Modelos Animais de Doenças , Pós-Condicionamento Isquêmico , Pulmão , Patologia , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Ratos Sprague-Dawley , Traumatismo por Reperfusão , Patologia , Proteína X Associada a bcl-2 , Metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate the role and significance of ATP-sensitive K+ channels in the pathological process of hypoxia hypercapnia-induced pulmonary vasoconstriction (HHPV) and the relationship with ERK1/2 signal pathway in rats.</p><p><b>METHODS</b>We made the third pulmonary artery rings of SD rats, used the model of pulmonary artery rings perfusion in vitro. Under acute hypoxia hypercapnia condition, and observed the effects of the three stages of HHPV incubated by glybenclamide(Gly) and the combined application of Gly and U0126. At the same time, the values of rings' tension changes were recorded via the method of hypoxia hypercapnia conditions reactivity.</p><p><b>RESULTS</b>Under the normoxia condition, the values of the third pulmonary artery rings tension were relatively stable, but under the hypoxia hypercapnia condition, we observed a biphasic pulmonary artery contractile response compared with N group (P < 0.05, P < 0.01). When the third pulmonary artery rings incubated by Gly, it's phase II persistent vasoconstriction was enhanced compared with the H group (P < 0.05, P < 0.01), and the phase I vasoconstriction was also heightened. Moreover, under the hypoxia hypercapnia condition, U0126 could significantly relieve the phase II persistent vasoconstriction compared with HD group (P < 0.05, P < 0.01) induced by Gly, but the phase I acute vasoconstriction and the phase I vasodilation had no changes (P > 0.05).</p><p><b>CONCLUSION</b>Gly may mediate HHPV via activating ERK1/2 signal transduction pathway.</p>
Assuntos
Animais , Masculino , Ratos , Glibureto , Farmacologia , Hipercapnia , Metabolismo , Hipóxia , Metabolismo , Técnicas In Vitro , Sistema de Sinalização das MAP Quinases , Fisiologia , Artéria Pulmonar , Metabolismo , Fisiologia , Ratos Sprague-Dawley , VasoconstriçãoRESUMO
<p><b>OBJECTIVE</b>To investigate the effects of ischemic postconditioning (IPostC) on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats.</p><p><b>METHODS</b>Adult male SD rats were randomly divided into 3 groups based upon the intervention (n = 8): control group (C), lung ischemic reperfusion group (LIR), LIR+ IPostC group (IPostC). At the end of the experiment, blood specimens drawn from the arteria carotis were tested for the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO); the pneumocyte apoptosis index (AI) was achieved by tennrminal deoxynucleotidyl transferase mediated dUTP nick end abeling (TUNEL); the expression of Bcl-2, Bax protein in lung tissue was accessed by quantitative immunohistochemistry (MHC) and Bcl-2, Bax mRNA by RT-PCR.</p><p><b>RESULTS</b>IPostC could significantly attenuate the MDA level, MPO activity and improve SOD activity in blood serum which was comparable to I/R and significantly reduced the number of TUNEL-positive cells compared with I/R group, expressed as Al (% total nuclei) from (39.0 +/- 3.46) to (8.0 +/- 0.88) (P < 0.01). The protein and mRNA expression of Bcl-2 and Bax showed that IPO significantly attenuated the ischemia/reperfusion-upregulated expression of Bax protein but improved the expression of Bcl-2 that improved the Bcl-2/Bax ratio (P < 0.01) .</p><p><b>CONCLUSION</b>IPostC may attenuate pneumocyte apoptosis in LIRI by up-regulating expression of Bcl-2/Bax ratio and by inhibiting oxidant generation and neutrophils filtration.</p>
Assuntos
Animais , Masculino , Ratos , Células Epiteliais Alveolares , Biologia Celular , Apoptose , Pós-Condicionamento Isquêmico , Pulmão , Metabolismo , Patologia , Lesão Pulmonar , Malondialdeído , Metabolismo , Peroxidase , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Ratos Sprague-Dawley , Traumatismo por Reperfusão , Superóxido Dismutase , Metabolismo , Proteína X Associada a bcl-2 , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of siRNA silencing the role of C-Jun N-terminal Kinase (JNK) gene in excessive endoplasmic reticulum stress on lung ischemia/reperfusion injury.</p><p><b>METHODS</b>Mouse model of pulmonary ischemia reperfusion injury (PIRI) in situ was established with unilateral lung in vivo. Seventy experimental mice were randomly allocated into seven groups (n = 10): Sham group (Sham group), ischemia reperfusion group (I/R), PBS+ Lipofectamine2000TM transfection reagent group (I/R + PBS+ Lipo group), negative control group (I/R+ SCR group), JNK-siRNA group (I/R + siRNA(JNK1), siRNA(JNK2), siRNA(JNK3)). Mice were euthanized after experimental time out, and left lung tissue was extracted. Wet/dry lung weight ratio (W/D) and total lung water content (TLW) were tested. Light microscope, alveolar damage quantitative evaluation index (IQA) and electron microscope were observed. The expression levels of JNK and glucose regulatex protein(GRP78) were detected by RT-PCR and Western blot. Apoptosis of lung tissue was determined by TUNEL.</p><p><b>RESULTS</b>Compared with Sham group, all indicators above of I/R + PBS + Lipo group and I/R + SCR group were significantly increased (P < 0.01), and compared with I/R group, those indicators of the three groups all had no notable difference; those indicators were not statistically different between I/R + PBS + Lipo group and I/R + SCR group, and compared to the three groups, the above indicators in JNK-siRNA group were lower (P < 0.05, P < 0.01) except that the expression levels of GRP78 was not statistically different.</p><p><b>CONCLUSION</b>I/R induces excessive ERS in lung tissue, in which JNK pathway participates in apoptosis, leading to lung tissue injury.</p>
Assuntos
Animais , Camundongos , Apoptose , Estresse do Retículo Endoplasmático , Proteínas de Choque Térmico , Metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno , Genética , Pulmão , Lesão Pulmonar , Genética , Sistema de Sinalização das MAP Quinases , RNA Interferente Pequeno , Traumatismo por Reperfusão , GenéticaRESUMO
<p><b>OBJECTIVE</b>To explore the role of Xuebijing Injection (XBJI) in inhibiting inflammatory factors associated with anoxia/reoxygenation myocardial inflammatory response of rats.</p><p><b>METHODS</b>Totally 36 healthy male Sprague-Dawley rats, 280 ± 30 g were randomly divided into six groups, i.e., the normal control group (N group), the balanced perfusion group (BP group),the model group (M group),the low dose XBJI group (XBJI(L) group), the middle dose XBJI group (XBJI(M) group),and the high dose XBJI group (XBJI(H) group), 6 in each group. The myocardial anoxia/reoxygenation rat model was established by Langendorff isolated heart perfusion. The concentration of TNF-α in the myocardial tissue was detected by ELISA. The expression of nuclear factor kappa B p65 (NF-κB p65) protein and Toll like receptor 4 (TLR4) protein were detected using Western blot. The expression of NF-κB p65 mRNA and TLR4 mRNA was detected by RT-PCR. Ultrastructural changes of anoxia-reoxygenation rats' heart muscle were observed under transmission electron microscope.</p><p><b>RESULTS</b>Compared with the M group,the TNF-α concentration, expression levels of NF-κB p65 protein and mRNA, TLR4 protein and mRNA decreased to various degrees in the XBJI(L) group, the XBJI(M) group, and the XBJI(H) group. The TNF-α expression level decreased most significantly in the XBJI(L), group (P < 0.01), while other indices decreased most obviously in the XBJI(M) group (P < 0.01, P < 0.05). Expression levels of NF-κB p65 and TLR4 protein were obviously lower in the XBJI(M) group than in the XBJI(L) group (P < 0.05). There was no statistical difference in other indices among the three XBJI groups (P > 0.05). Myocardial fibers were loose and broken with disappearance of transverse striation, and mitochondrial cristae was dissolved and severely damaged in the M group. The aforesaid condition was improved after treated by XBJI, with the most obvious effect obtained in the XBJI(M) group.</p><p><b>CONCLUSIONS</b>Different doses of XBJI could attenuate inflammatory reactions after myocardial anoxia/reoxygenation rats' heart muscle through inhibiting TLR4-NF-κB-TNF-α signal transduction pathway. The best effect could be obtained by 4 mL/100 mL XBJI.</p>
Assuntos
Animais , Masculino , Ratos , Medicamentos de Ervas Chinesas , Farmacologia , Usos Terapêuticos , Hipóxia , Miocárdio , Metabolismo , Miócitos Cardíacos , NF-kappa B , Metabolismo , Oxigênio , Metabolismo , RNA Mensageiro , Ratos Sprague-Dawley , Transdução de Sinais , Receptor 4 Toll-Like , Metabolismo , Fator de Transcrição RelA , Fator de Necrose Tumoral alfa , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To explore the effect of curcumin (CUR) on cycteinyl aspirate specific protease-12 (Caspase-12) and pneumocyte apoptosis in pulmonary ischemia/reperfusion (I/R) injury mice.</p><p><b>METHODS</b>The in vivo unilateral in situ pulmonary I/R injury mouse model was established in C57BL/6J mice. Sixty experimental mice were randomly divided into six groups by random digit table, i. e., the sham-operation group (Sham), the I/R group, the I/R + dimethyl sulfoxide group (I/R + DMSO), the I/R + low dose CUR pre-treated group (I/R + CUR-100), the I/R + middle dose CUR pre-treated group (I/R + CUR-150), the I/R + high dose CUR pre-treated group (I/R + CUR-200), 10 in each group. Mice were euthanized and their left lungs were excised. Wet lung weight to dry lung weight (W/D) and the total lung water content (TLW) were tested. The morphological changes of the lung tissue were observed and index of quantitative evaluation for alveolar damage (IQA) detected under light microscope. The ultra-microstructure of the lung tissue was observed under electron microscope. The mRNA and protein expression levels of Caspase-12 and glucose regulated protein (GRP78) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Apoptosis index (AI) of the lung tissue was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>Compared with the Sham group, expression levels of Caspase-12, GRP78 mRNA and protein all significantly increased in the I/R group (P < 0.05); W/D, TLW, IQA, and AI were all notably higher (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were notably observed in I/R group. Compared with the I/R + DMSO group, expression levels of GRP78 mRNA and protein were increasingly higher in the I/R + CUR-100 group, the I/R + CUR-150 group, and the I/R +CUR-200 group (P < 0.05), expression levels of Caspase-12 mRNA and protein were lower (P < 0.05); W/D, TLW, IQA, and AI also decreased (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were gradually alleviated in the I/R + CUR groups.</p><p><b>CONCLUSION</b>CUR had better effect on the lung protection against I/R injury, which might be related to inhibition for pneumocyte apoptosis associated with Caspase-12 in excessive unfolded protein response (UPR).</p>
Assuntos
Animais , Masculino , Camundongos , Apoptose , Caspase 12 , Metabolismo , Curcumina , Farmacologia , Proteínas de Choque Térmico , Metabolismo , Pulmão , Camundongos Endogâmicos C57BL , Traumatismo por Reperfusão , Metabolismo , PatologiaRESUMO
OBJECTIVE: To investigate the effects of P38MAPK on erythropoietin (EPO) postconditioning attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI) in rats. METHODS: Adult male Sprague-Dawley rats were randomly divided into 5 groups based upon the intervention (n=8); control group (C), LIRI group (I/R), LIRI+EPO group (EPO), EPO+solution countrl group (D), EPO+SB203580 group (SB). At the end of the experiment, blood specimens drawn from the arteria carotis were tested for the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO). The histological structure of the left lung was observed under light microscope, and scored by alveolar damage index of quantitative assessment (IQA). The pneumocyte apoptosis index (AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end a-beling (TUNEL). RESULTS: Compared with C group, in I/R group IQA, AI and MDA level, MPO activity were significantly increased, SOD activity was reduced (P<0.05 or P<0.01), and morphological abnormality occurred in lung tissue. Compared with 1/R group, IQA, AI and MDA level, MPO activity were significantly decreased, SOD activity was improved (P<0.05 or P<0.01), and morphological abnormality in lung tissue was obviously reduced in EPO, D and SB groups. There were no significant difference in all of the indexes between D and EPO groups (P≤0.05). However, compared with EPO group, SOD activity was increased (P<0.05 or P<0.01), the other indexes were obviously reduced, and the abnormal changes of the morphology in I/R were also improved markedly in SB group. CONCLUSION: EPO may attenuate pneumocyte apoptosis in LIRI by reducing oxidant generation, neutrophils filtration, then inhibiting activation of P38MAPK.
RESUMO
<p><b>OBJECTIVE</b>To investigate the effects of curcumin (CUR) on pneumocyte apoptosis and CCAAT/enhancer binding protein homologous protein (CHOP) in pulmonary ischemia/reperfusion injury (PIRI) in mice.</p><p><b>METHODS</b>Sixty C57BL/6J mice were randomly allocated into six groups (n = 10): Sham operation group (Sham group), ischemia/reperfusion group (I/R group), ischemia/reperfusion + dimethyl sulfoxide group (DMSO group), ischemia/reperfusion + curcumin pre-treated with respectively 100 mg/kg, 150 mg/kg and 200 mg/kg groups (CUR-100 group, CUR-150 group and CUR-200 group). Left lung tissue of each group was excised after reperfusion for 3 h. Wet lung weight to dry lung weight (W/D) and total lung water content (TLW) were tested. The morphological and ultrastructural changes of lung tissue were observed under light microscope and electron microscope, and index of quantitative evaluation for alveolar damage (IQA) was calculated. The expression levels of CHOP and glucose regulated protein 78 (GRP78) were detected by RT-PCR and Western Blot. Apoptosis index (AI) of lung tissue was determined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>Compared with Sham group, the expression levels of CHOP, GRP78 mRNA and protein were all significantly increased (P < 0.05) in I/R group and DMSO group, W/D, TLW, IQA and AI were all notably higher (P < 0.01); morphological and ultrastructural injury in lung tissue were notably observed in I/R group. Compared with DMSO group, the expression levels of GRP78 mRNA and protein were increased higher (P < 0. 05) in CUR-100 group, CUR-150 group, and CUR-200 group, but the expression levels of CHOP mRNA and protein were decreased lower (P < 0.05), W/D, TLW, IQA and AI were also decreased (P < 0.05, P < 0.01); morphological and ultrastructural injury in lung tissue were gradually alleviated in CUR groups.</p><p><b>CONCLUSION</b>I/R induces excessive unfolded protein response (UPR) in lung tissue, in which CHOP participates in pneumocyte apoptosis, leading to lung injury; CUR has notable effects on lung protection against I/R injury, which may be related to inhibition of apoptosis mediated by CHOP in excessive UPR.</p>
Assuntos
Animais , Masculino , Camundongos , Células Epiteliais Alveolares , Metabolismo , Apoptose , Curcumina , Farmacologia , Proteínas de Choque Térmico , Metabolismo , Pulmão , Metabolismo , Patologia , Camundongos Endogâmicos C57BL , Traumatismo por Reperfusão , Metabolismo , Patologia , Fator de Transcrição CHOP , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of diabetic autonomic neuropathy on the seminal vesicle and search for the theoretical evidence for the prevention and treatment of diabetic infertility by observing changes in the contents of the nerve growth factor (NGF) and muscarinic M3 receptor in the seminal vesicle of diabetic rats.</p><p><b>METHODS</b>Diabetic models were established in 10 of the 15 male adult SD rats by intraperitoneal injection of streptozotocin (STZ), and the other 5 were included in a normal control group. Eight weeks after modeling, seminal vesicles were collected from the rats for HE and immunohistochemical staining.</p><p><b>RESULTS</b>Compared with the normal controls, the diabetic models showed a decreased number of smooth muscle cells, thinner cytoplasm of glandular epithelial cells and disordered structure in the seminal vesicle. The intensity of NGF-positive staining was significantly enhanced, but that of M3 markedly reduced in the diabetic group. There were statistically significant differences in the mean integrated optical density (IA) of muscarinic M3 receptors and NGF between the control and diabetic groups (0.0187 +/- 0.0024 vs 0.0100 +/- 0.0015 and 0.0209 +/- 0.0085 vs 0.0412 +/- 0.0117, P<0.01).</p><p><b>CONCLUSION</b>The changes in the expressions of NGF and M3 receptors in the seminal vesicle of diabetic rats suggest that diabetes mellitus may induce autonomic neuropathy of the seminal vesicle.</p>