AIM: To determine the in vivo andin vivo effects of cysteamine (CS) on expression and activity of H+-K+-ATPase of gastric mucosal cells in weaning piglets.METHODS: Eighteen litters of newborn Xinhuai piglets were empl...AIM: To determine the in vivo andin vivo effects of cysteamine (CS) on expression and activity of H+-K+-ATPase of gastric mucosal cells in weaning piglets.METHODS: Eighteen litters of newborn Xinhuai piglets were employed in the in vivo experiment and allocated to control and treatment groups. From 12 d of age (D12), piglets in control group were fed basal diet, while the treatment group received basal diet supplemented with 120 mg/kg CS. Piglets were weaned on D35 in both groups. Six piglets from each group (n = 6) were slaughtered on D28 (one week before weaning), D35(weaning), D36.5, D38, D42, and D45 (36 h, 72 h,one week and 10 d after weaning), respectively. Semiquantitative RT-PCR was performed todetermine the levels of H+-K+-ATPase mRNA in gastric mucosa. H+-K+-ATPase activity in gastric mucosa homogenate was also determined. Gastric mucosal epithelial cells from piglets through primary cultures were used to further elucidate the effect of CS on expression and activity of H+-K+-ATPase in vivo. Cells were treated for 20 h with 0.001,0.01, and 0.1 mg/mL of CS (n = 4), respectively. The mRNA expression of H+-K+-ATPase and somatostatin (SS)as well as the H+-K+-ATPase activity were determined.RESULTS: in vivo, both mRNA expression and activity of H+-K+-ATPase in gastric mucosa of control group exhibited a trend to increase from D28 to D45, reaching a peak on D45, but did not show significant age differences. Furthermore, neither the mRNA expression nor the activity of H+-K+-ATPase was affected significantly by weaning. CS increased the mRNA expression of H+-K+-ATPase by 73%, 53%, 30% and 39% on D28(P = 0.014), D35 (P = 0.017), D42 (P = 0.013) and D45(P = 0.046), respectively. In accordance with the mRNA expression, H+-K+-ATPase activities were significantly higher in treatment group than in control group on D35(P = 0.043) and D45 (P = 0.040). In vivo, CS exhibited a dose-dependent effect on mRNA expression and activity of H+-K+-ATPase. Both H+-K+-ATPase mRNA expression and activity in gastric mucosal epithelial cells were significantly elevated after 20 h of exposure to the moderate (H+-K+-ATPase expression: P=0.03; H+-K+-ATPase activity: P = 0.014) and high concentrations (H+-K+-ATPase expression: P=0.017; H+-K+-ATPase activity:P = 0.022) of CS. Significant increases in SS mRNA expression were observed to accompany the elevation of H+-K+-ATPase expression and activity induced by the moderate (P = 0.024) and high concentrations (P = 0.022) of CS. Low concentration of CS exerted no effects either on expression and activity of H+-K+-ATPase or on SS mRNA expression in cultured gastric mucosal epithelial cells.CONCLUSION: No significant changes are observed in mRNA expression and activity of H+-K+-ATPase in gastric mucosa of piglets around weaning from D28 to D45. CS increases expression and activity of gastric H+-K+-ATPase in vivo and in vivo. SS is involved in mediating the effect of CS on gastric H+-K+-ATPase expression and activity in weaning piglets.展开更多
Plasma membrane （PM） Ca2+-ATPase activity in poplar apical bud meristematic cells during short-day （SD）-induced dormancy development was examined by a cerium precipitation EM-cytochemical method. Ca2+-ATPase activ...Plasma membrane （PM） Ca2+-ATPase activity in poplar apical bud meristematic cells during short-day （SD）-induced dormancy development was examined by a cerium precipitation EM-cytochemical method. Ca2+-ATPase activity, indicated by the status of cerium phosphate precipitated grains, was localized mainly on the interior face （cytoplasmic side） of the PM when plants were grown under long days and reached a deep dormancy. A few reaction products were also observed on the nuclear envelope.When plant buds were developing dormancy after 28 to 42 d of SD exposure, almost no reaction products were present on the interior face of the PM. In contrast, a large number of cerium phosphate precipitated grains were distributed on the exterior face of the PM. After 70 d of SD exposure, when buds had developed a deep dormancy, the reaction products of Ca2+-ATPase activity again appeared on the interior face of the PM. The results seemed suggesting that two kinds of Ca2+-ATPases may be present on the PM during the SD-induced dormancy in poplar.One is the Ca2+-punlping ATPase, which is located on the interior face of the PM, for maintaining and restoring the Ca2+homeostasis. The other might be an ecto-Ca2+-ATPase, which is located on the exterior face of the PM, for the exocytosis of cell wall materials as suggested by the fact of the cell wall thickening during the dormancy developlnent in poplar.展开更多
The effects of ultradry storage on the activity of ATPase and fluidity of plasma membrane in Chinese cabbage (Brassica pekinensis (Lour.) Rupr.) and elm ( Ulmus pumila L.) seeds were investigated. The results indicate...The effects of ultradry storage on the activity of ATPase and fluidity of plasma membrane in Chinese cabbage (Brassica pekinensis (Lour.) Rupr.) and elm ( Ulmus pumila L.) seeds were investigated. The results indicated that no significant differences in the activity of ATPase and the micro- viscosity of plasma membrane of ultradried (UD) seeds could be found as compared to the control seeds stored under -20 degreesC, although there was a little adverse effect on the seeds with extreme dehydration. The results were consistent with higher vigor level of UD seeds. This implied that ultradry seed storage could protect the integrity of the membrane, maintain its physiological function and improve the storability of seeds. The relationship between sugar and desiccation tolerance of UD seeds was analysed using the HPLC. The results showed that the ratio of reducing to nonreducing sugar was lower in UD seeds than that in control. The content of sucrose and stachyose in elm and Chinese cabbage UD seeds was related to their desiccation tolerance, while no stachyose was detected in corn (Zea mays L.) seeds. This could be one of the reasons for its sensitivity to desiccation.展开更多
Objective: To investigate the changes of proton transportation across the inner mitochondrial membrane (IMM) and H + ATPase of hepatocytes in endotoxic shock rats. Methods: Endotoxin from E.Coil of 5.0 mg/kg or saline...Objective: To investigate the changes of proton transportation across the inner mitochondrial membrane (IMM) and H + ATPase of hepatocytes in endotoxic shock rats. Methods: Endotoxin from E.Coil of 5.0 mg/kg or saline of 1 ml/kg was injected into the femoral vein. The rats were sacrificed pre injection and 1, 3, 5, 8 hours after injection, and plasma and liver tissue samples were collected respectively. The liver tissue samples were used for preparation of mitochondria and submitochondrial particles (SMPs). The proton translocation of SMPs and H + ATPase, phospholipase A 2 (PLA 2) activities and malondialdehyde (MDA) content, membrane fluidities of different level of mitochondria membrane and plasma MDA content were assayed. Results: (1) Five hours after E. Coli. O111B4 injection, the maximum fluorescence quenching ACMA after adding ATP, nicotinamide adenin dinucleoacid hydrogen (NADH), and the succinate were significantly decreased (P< 0.05 ). The time of maximum fluorescent quenching and the half time of fluorescent quenching were significantly prolonged (P< 0.01 ), especially when NADH was used as a substrate. (2) The mitochondrial H + ATPase activity was significantly increased at early stage of endotoxic shock (P< 0.05 ), and significantly decreased at late stage of endotoxic shock (P< 0.01 ). (3) The mitochondrial membrane bound PLA 2 activity, plasmal and mitochondrial MDA content were significantly increased and succinate dehydrogenase (SDH) activity of mitochondria decreased markedly in endotoxic shock rats (P< 0.05 ). (4) The mitochondrial membrane fluidity of different lipid regions was decreased, especially in the head of phospholipid. Conclusions: Proton transportation across IMM and mitochondrial H + ATPase activity are significantly decreased in endotoxic shock.展开更多
基金Supported by the National Natural Science Foundation of China, No. 30270975 National Basic Research Program of China, No. 2004CB117505
文摘AIM: To determine the in vivo andin vivo effects of cysteamine (CS) on expression and activity of H+-K+-ATPase of gastric mucosal cells in weaning piglets.METHODS: Eighteen litters of newborn Xinhuai piglets were employed in the in vivo experiment and allocated to control and treatment groups. From 12 d of age (D12), piglets in control group were fed basal diet, while the treatment group received basal diet supplemented with 120 mg/kg CS. Piglets were weaned on D35 in both groups. Six piglets from each group (n = 6) were slaughtered on D28 (one week before weaning), D35(weaning), D36.5, D38, D42, and D45 (36 h, 72 h,one week and 10 d after weaning), respectively. Semiquantitative RT-PCR was performed todetermine the levels of H+-K+-ATPase mRNA in gastric mucosa. H+-K+-ATPase activity in gastric mucosa homogenate was also determined. Gastric mucosal epithelial cells from piglets through primary cultures were used to further elucidate the effect of CS on expression and activity of H+-K+-ATPase in vivo. Cells were treated for 20 h with 0.001,0.01, and 0.1 mg/mL of CS (n = 4), respectively. The mRNA expression of H+-K+-ATPase and somatostatin (SS)as well as the H+-K+-ATPase activity were determined.RESULTS: in vivo, both mRNA expression and activity of H+-K+-ATPase in gastric mucosa of control group exhibited a trend to increase from D28 to D45, reaching a peak on D45, but did not show significant age differences. Furthermore, neither the mRNA expression nor the activity of H+-K+-ATPase was affected significantly by weaning. CS increased the mRNA expression of H+-K+-ATPase by 73%, 53%, 30% and 39% on D28(P = 0.014), D35 (P = 0.017), D42 (P = 0.013) and D45(P = 0.046), respectively. In accordance with the mRNA expression, H+-K+-ATPase activities were significantly higher in treatment group than in control group on D35(P = 0.043) and D45 (P = 0.040). In vivo, CS exhibited a dose-dependent effect on mRNA expression and activity of H+-K+-ATPase. Both H+-K+-ATPase mRNA expression and activity in gastric mucosal epithelial cells were significantly elevated after 20 h of exposure to the moderate (H+-K+-ATPase expression: P=0.03; H+-K+-ATPase activity: P = 0.014) and high concentrations (H+-K+-ATPase expression: P=0.017; H+-K+-ATPase activity:P = 0.022) of CS. Significant increases in SS mRNA expression were observed to accompany the elevation of H+-K+-ATPase expression and activity induced by the moderate (P = 0.024) and high concentrations (P = 0.022) of CS. Low concentration of CS exerted no effects either on expression and activity of H+-K+-ATPase or on SS mRNA expression in cultured gastric mucosal epithelial cells.CONCLUSION: No significant changes are observed in mRNA expression and activity of H+-K+-ATPase in gastric mucosa of piglets around weaning from D28 to D45. CS increases expression and activity of gastric H+-K+-ATPase in vivo and in vivo. SS is involved in mediating the effect of CS on gastric H+-K+-ATPase expression and activity in weaning piglets.
文摘Plasma membrane （PM） Ca2+-ATPase activity in poplar apical bud meristematic cells during short-day （SD）-induced dormancy development was examined by a cerium precipitation EM-cytochemical method. Ca2+-ATPase activity, indicated by the status of cerium phosphate precipitated grains, was localized mainly on the interior face （cytoplasmic side） of the PM when plants were grown under long days and reached a deep dormancy. A few reaction products were also observed on the nuclear envelope.When plant buds were developing dormancy after 28 to 42 d of SD exposure, almost no reaction products were present on the interior face of the PM. In contrast, a large number of cerium phosphate precipitated grains were distributed on the exterior face of the PM. After 70 d of SD exposure, when buds had developed a deep dormancy, the reaction products of Ca2+-ATPase activity again appeared on the interior face of the PM. The results seemed suggesting that two kinds of Ca2+-ATPases may be present on the PM during the SD-induced dormancy in poplar.One is the Ca2+-punlping ATPase, which is located on the interior face of the PM, for maintaining and restoring the Ca2+homeostasis. The other might be an ecto-Ca2+-ATPase, which is located on the exterior face of the PM, for the exocytosis of cell wall materials as suggested by the fact of the cell wall thickening during the dormancy developlnent in poplar.
文摘The effects of ultradry storage on the activity of ATPase and fluidity of plasma membrane in Chinese cabbage (Brassica pekinensis (Lour.) Rupr.) and elm ( Ulmus pumila L.) seeds were investigated. The results indicated that no significant differences in the activity of ATPase and the micro- viscosity of plasma membrane of ultradried (UD) seeds could be found as compared to the control seeds stored under -20 degreesC, although there was a little adverse effect on the seeds with extreme dehydration. The results were consistent with higher vigor level of UD seeds. This implied that ultradry seed storage could protect the integrity of the membrane, maintain its physiological function and improve the storability of seeds. The relationship between sugar and desiccation tolerance of UD seeds was analysed using the HPLC. The results showed that the ratio of reducing to nonreducing sugar was lower in UD seeds than that in control. The content of sucrose and stachyose in elm and Chinese cabbage UD seeds was related to their desiccation tolerance, while no stachyose was detected in corn (Zea mays L.) seeds. This could be one of the reasons for its sensitivity to desiccation.
文摘Objective: To investigate the changes of proton transportation across the inner mitochondrial membrane (IMM) and H + ATPase of hepatocytes in endotoxic shock rats. Methods: Endotoxin from E.Coil of 5.0 mg/kg or saline of 1 ml/kg was injected into the femoral vein. The rats were sacrificed pre injection and 1, 3, 5, 8 hours after injection, and plasma and liver tissue samples were collected respectively. The liver tissue samples were used for preparation of mitochondria and submitochondrial particles (SMPs). The proton translocation of SMPs and H + ATPase, phospholipase A 2 (PLA 2) activities and malondialdehyde (MDA) content, membrane fluidities of different level of mitochondria membrane and plasma MDA content were assayed. Results: (1) Five hours after E. Coli. O111B4 injection, the maximum fluorescence quenching ACMA after adding ATP, nicotinamide adenin dinucleoacid hydrogen (NADH), and the succinate were significantly decreased (P< 0.05 ). The time of maximum fluorescent quenching and the half time of fluorescent quenching were significantly prolonged (P< 0.01 ), especially when NADH was used as a substrate. (2) The mitochondrial H + ATPase activity was significantly increased at early stage of endotoxic shock (P< 0.05 ), and significantly decreased at late stage of endotoxic shock (P< 0.01 ). (3) The mitochondrial membrane bound PLA 2 activity, plasmal and mitochondrial MDA content were significantly increased and succinate dehydrogenase (SDH) activity of mitochondria decreased markedly in endotoxic shock rats (P< 0.05 ). (4) The mitochondrial membrane fluidity of different lipid regions was decreased, especially in the head of phospholipid. Conclusions: Proton transportation across IMM and mitochondrial H + ATPase activity are significantly decreased in endotoxic shock.
