Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspa...Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspase-3) in CCl4-induced hepatic fibrosis in rats. And also it is to explore the mechanism of DSHX in anti-fibrosis. Methods: Eighty male Wistar rats were randomly divided into the normal control group (A), the model group (B), the un-treated model group (C), the low-dose-DSHX treated group (D) and the high-dose-DSHX treated group (E). Except those in Group A, all the other rats were made into hepatic fibrotic models by comprehensive processes including subcutaneous injecting of CCl4, and feeding them with alcoholic high-fat and low-protein diet for 8 weeks. Then the two DSHX-treated groups were treated respectively with low dose (0.5 g/kg) and high dose (1.0 g/kg) DSHX capsule by gastrogavage everyday for 8 weeks. At the end of the experiment, the liver index, levels of hyaluronic acid (HA) and alanine amin-otransferase (ALT) in serum, degree of hepatic fibrosis, and urinary excretion of hydroxyproline (Hyp) were measured, and the expression of Col Ⅰ , Col Ⅲ and caspase-3 in liver tissues were detected respectively by immunohistochemistric technique. Results: Compared with those in Group B and C, the two DSHX treated groups showed that the liver index, levels of serum HA and ALT and severity of hepatic fibrosis were all significantly lower, the urinary excretion of Hyp was significantly higher; the Col Ⅰ and Col-Ⅲ expression was lower (Col Ⅰ :1. 23±1.14,1. 07±0. 96 vs 4.18±2. 26, 3. 22±1. 44, P<0. 01;Col Ⅲ : 1. 31±0. 69, , 1. 09± 0.58 vs 3.04±0.62,2.23±0.58, P<0.05). At the same time, the expression of caspase-3 in Group E was fewer than Group B and C in hepatocytes (3. 09±0. 65 vs 9. 60±2. 32, 8. 82 ±1. 45, P<0.01),but it was extensively expressed in fibrous septal cells(4.52±0.87 vs 1.69±0.23,2.98±0.36, P<0.01). Conclusion: DSHX capsule shows certain therapeutic effect on hepatic fibrosis in rats, and the mechanism might be related with reducing Col Ⅰ and Col Ⅲ deposition, inhibiting hepatocyte apoptosis and promoting fibrous septal cells (mainly the activated hepatic stellate cells) apoptosis.展开更多
Smoking during pregnancy is a major source of fetal exposure to numerous harmful agents present in tobacco smoke. Lung development involves complex biochemical processes resulting in dramatic changes which continue ev...Smoking during pregnancy is a major source of fetal exposure to numerous harmful agents present in tobacco smoke. Lung development involves complex biochemical processes resulting in dramatic changes which continue even after birth. In addition to type I cells which form the blood-air barrier, type II alveolar epithelial (AE) cells have important and diverse functions related to immunological protection and stabilization of the alveolus through synthesis and secretion of the pulmonary surfactant. Apoptosis or programmed cells death is an important physiological process during lung embryogenesis and for the proper maintenance of homeostasis. Caspases are proteases that play important roles in regulating apoptosis. Caspase-3 is the key executioner caspase in the cascade of events leading to cell death by apoptosis. We explored the hypothesis that cigarette smoke extract (CSE) induces apoptosis in fetal rat lung type II AE cells by activation of caspase-3. To analyze these factors, isolated fetal rat lung type II AE cells were used. The cells were exposed to different concentrations of CSE (5%, 10% or 15%) (v/v) for 60 min. The results of the present study showed that CSE induced apoptosis in fetal rat lung type II AE cells with a significant increase (p 0.05) in caspase-3 activity and decrease in cell proliferation at CSE concentrations of 10% and 15% (v/v). These observations indicate that cigarette smoke extract induces apoptosis by activation of caspase-3 in fetal rat lung type II AE cells in a dose-dependent manner and may potentially alter the regulated development of the lung and the appearance of the surfactant-producing type II alveolar cells which are critical for the establishment of adequate gas exchange at birth.展开更多
Enterovirus A71(EV-A71) is the major pathogen responsible for the severe hand, foot and mouth disease worldwide, for which few effective antiviral drugs are presently available. Interferon-a(IFN-a) has been used in an...Enterovirus A71(EV-A71) is the major pathogen responsible for the severe hand, foot and mouth disease worldwide, for which few effective antiviral drugs are presently available. Interferon-a(IFN-a) has been used in antiviral therapy for decades;it has been reported that EV-A71 antagonizes the antiviral activity of IFN-a based on viral 2 Apro-mediated reduction of the interferon-alpha receptor 1(IFNAR1);however, the mechanism remains unknown. Here, we showed a significant increase in IFNAR1 protein induced by IFN-a in RD cells, whereas EV-A71 infection caused obvious downregulation of the IFNAR1 protein and blockage of IFN-a signaling. Subsequently, we observed that EV-A71 2 Apro inhibited IFNAR1 translation by cleavage of the eukaryotic initiation factor 4 GI(eIF4GI), without affecting IFNAR1 m RNA levels induced by IFN-a. The inhibition of IFNAR1 translation also occurred in puromycin-induced apoptotic cells when caspase-3 cleaved e IF4 GI. Importantly, we verified that 2 Aprocould activate cellular caspase-3, which was subsequently involved in e IF4 GI cleavage mediated by 2 Apro. Furthermore, inhibition of caspase-3 activation resulted in the partial restoration of IFNAR1 in cells transfected with 2 A or infected with EV-A71, suggesting the pivotal role of both viral 2 Aproand caspase-3 activation in the disturbance of IFN-a signaling. Collectively, we elucidate a novel mechanism by which cellular caspase-3 contributes to viral 2 Apro-mediated down-regulation of IFNAR1 at the translation level during EV-A71 infection, indicating that caspase-3 inhibition could be a potential complementary strategy to improve clinical anti-EV-A71 therapy with IFN-a.展开更多
文摘Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspase-3) in CCl4-induced hepatic fibrosis in rats. And also it is to explore the mechanism of DSHX in anti-fibrosis. Methods: Eighty male Wistar rats were randomly divided into the normal control group (A), the model group (B), the un-treated model group (C), the low-dose-DSHX treated group (D) and the high-dose-DSHX treated group (E). Except those in Group A, all the other rats were made into hepatic fibrotic models by comprehensive processes including subcutaneous injecting of CCl4, and feeding them with alcoholic high-fat and low-protein diet for 8 weeks. Then the two DSHX-treated groups were treated respectively with low dose (0.5 g/kg) and high dose (1.0 g/kg) DSHX capsule by gastrogavage everyday for 8 weeks. At the end of the experiment, the liver index, levels of hyaluronic acid (HA) and alanine amin-otransferase (ALT) in serum, degree of hepatic fibrosis, and urinary excretion of hydroxyproline (Hyp) were measured, and the expression of Col Ⅰ , Col Ⅲ and caspase-3 in liver tissues were detected respectively by immunohistochemistric technique. Results: Compared with those in Group B and C, the two DSHX treated groups showed that the liver index, levels of serum HA and ALT and severity of hepatic fibrosis were all significantly lower, the urinary excretion of Hyp was significantly higher; the Col Ⅰ and Col-Ⅲ expression was lower (Col Ⅰ :1. 23±1.14,1. 07±0. 96 vs 4.18±2. 26, 3. 22±1. 44, P<0. 01;Col Ⅲ : 1. 31±0. 69, , 1. 09± 0.58 vs 3.04±0.62,2.23±0.58, P<0.05). At the same time, the expression of caspase-3 in Group E was fewer than Group B and C in hepatocytes (3. 09±0. 65 vs 9. 60±2. 32, 8. 82 ±1. 45, P<0.01),but it was extensively expressed in fibrous septal cells(4.52±0.87 vs 1.69±0.23,2.98±0.36, P<0.01). Conclusion: DSHX capsule shows certain therapeutic effect on hepatic fibrosis in rats, and the mechanism might be related with reducing Col Ⅰ and Col Ⅲ deposition, inhibiting hepatocyte apoptosis and promoting fibrous septal cells (mainly the activated hepatic stellate cells) apoptosis.
文摘Smoking during pregnancy is a major source of fetal exposure to numerous harmful agents present in tobacco smoke. Lung development involves complex biochemical processes resulting in dramatic changes which continue even after birth. In addition to type I cells which form the blood-air barrier, type II alveolar epithelial (AE) cells have important and diverse functions related to immunological protection and stabilization of the alveolus through synthesis and secretion of the pulmonary surfactant. Apoptosis or programmed cells death is an important physiological process during lung embryogenesis and for the proper maintenance of homeostasis. Caspases are proteases that play important roles in regulating apoptosis. Caspase-3 is the key executioner caspase in the cascade of events leading to cell death by apoptosis. We explored the hypothesis that cigarette smoke extract (CSE) induces apoptosis in fetal rat lung type II AE cells by activation of caspase-3. To analyze these factors, isolated fetal rat lung type II AE cells were used. The cells were exposed to different concentrations of CSE (5%, 10% or 15%) (v/v) for 60 min. The results of the present study showed that CSE induced apoptosis in fetal rat lung type II AE cells with a significant increase (p 0.05) in caspase-3 activity and decrease in cell proliferation at CSE concentrations of 10% and 15% (v/v). These observations indicate that cigarette smoke extract induces apoptosis by activation of caspase-3 in fetal rat lung type II AE cells in a dose-dependent manner and may potentially alter the regulated development of the lung and the appearance of the surfactant-producing type II alveolar cells which are critical for the establishment of adequate gas exchange at birth.
基金grants from Beijing Natural Science Foundation(No.19G10290)National Natural Science Foundation of China(No.81772184).
文摘Enterovirus A71(EV-A71) is the major pathogen responsible for the severe hand, foot and mouth disease worldwide, for which few effective antiviral drugs are presently available. Interferon-a(IFN-a) has been used in antiviral therapy for decades;it has been reported that EV-A71 antagonizes the antiviral activity of IFN-a based on viral 2 Apro-mediated reduction of the interferon-alpha receptor 1(IFNAR1);however, the mechanism remains unknown. Here, we showed a significant increase in IFNAR1 protein induced by IFN-a in RD cells, whereas EV-A71 infection caused obvious downregulation of the IFNAR1 protein and blockage of IFN-a signaling. Subsequently, we observed that EV-A71 2 Apro inhibited IFNAR1 translation by cleavage of the eukaryotic initiation factor 4 GI(eIF4GI), without affecting IFNAR1 m RNA levels induced by IFN-a. The inhibition of IFNAR1 translation also occurred in puromycin-induced apoptotic cells when caspase-3 cleaved e IF4 GI. Importantly, we verified that 2 Aprocould activate cellular caspase-3, which was subsequently involved in e IF4 GI cleavage mediated by 2 Apro. Furthermore, inhibition of caspase-3 activation resulted in the partial restoration of IFNAR1 in cells transfected with 2 A or infected with EV-A71, suggesting the pivotal role of both viral 2 Aproand caspase-3 activation in the disturbance of IFN-a signaling. Collectively, we elucidate a novel mechanism by which cellular caspase-3 contributes to viral 2 Apro-mediated down-regulation of IFNAR1 at the translation level during EV-A71 infection, indicating that caspase-3 inhibition could be a potential complementary strategy to improve clinical anti-EV-A71 therapy with IFN-a.
