目的:研究孕酮与钙离子载体A23187诱导顶体反应(Acrosom e reaction,AR)特征。方法:10例健康生育男性精液,采用Percoll密度梯度分离法优选精子,采用FITC-PSA荧光染色法分析AR。结果:A23187诱导AR类型主要为III型和IV型,孕酮诱导AR类型...目的:研究孕酮与钙离子载体A23187诱导顶体反应(Acrosom e reaction,AR)特征。方法:10例健康生育男性精液,采用Percoll密度梯度分离法优选精子,采用FITC-PSA荧光染色法分析AR。结果:A23187诱导AR类型主要为III型和IV型,孕酮诱导AR类型主要为II型。精子获能4、6、8和16 h时,A23187和孕酮诱导AR率与获能0 h组相比较,均明显增加,两组比较差异均有显著性(P<0.05),同一获能时间A 23187和孕酮诱导的AR率比较,差异均无显著性(P>0.05)。A23187诱导15 m in、30 m in和60 m in时,AR率均明显高于诱导前AR率(P<0.005)。孕酮诱导15 m in时,AR率与诱导前组比较没有明显差异(P>0.05),孕酮诱导30 m in和60 m in时,AR率均明显高于诱导前组(P<0.005),同一诱导时间,A23187诱导AR率均明显高于孕酮诱导的AR率,差异均有显著性(P<0.05)。结论:孕酮可诱导精子发生AR,诱导AR类型主要为II型,可比A23187诱导的AR更接近生理过程。展开更多
The aim of the study was to prepare berberine hydrochloride long-circulating liposomes and optimize the formulation and process parameters,and investigate the influence of different factors on the encapsulation effici...The aim of the study was to prepare berberine hydrochloride long-circulating liposomes and optimize the formulation and process parameters,and investigate the influence of different factors on the encapsulation efficiency.Berberine hydrochloride liposomes were prepared in response to a transmembrane ion gradient that was established by ionophore A23187.Free and liposomal drug were separated by cation exchange resin,and then the amount of intraliposomal berberine hydrochloride was determined by UV spectrophotometry.The optimized encapsulation efficiency of berberine hydrochloride liposomes was 94.3%2.1%when the drug-to-lipid ratio was 1:20,and the mean diameter was 146.9 nm3.2 nm.As a result,the ionophore A23187-mediated ZnSO_(4)gradient method was suitable for the preparation of berberine hydrochloride liposomes that we could get the desired encapsulation efficiency and drug loading.展开更多
Effects of procainamide (PA) on human platelet aggregation and the cytosolic free-Ca2+ concentration ([Ca2+]) were investigated in vitro. PA at doses of 8.5 , 34 and 136 μmol·L-1 could inhibit the human platelet...Effects of procainamide (PA) on human platelet aggregation and the cytosolic free-Ca2+ concentration ([Ca2+]) were investigated in vitro. PA at doses of 8.5 , 34 and 136 μmol·L-1 could inhibit the human platelet aggregation induced by 0. 5 μmol · L-1 A23187 with a good concentration -effect relationship. One min and maximal aggregation rates were also inhibited ( P<0. 01 ,vs control). The [Ca2+], was decreased in the presence of PA ,and the changes of [Ca2+], showed a significant linear correlation with 1 min or maximal aggregation rate (P<0.05). These results suggest that the mechanisms of PA inhibiting platelet aggregation relate to the decrease of [Ca2+].展开更多
Objective To investigate the effects of endothelial microvesicles(EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes. Methods Human umbilical vein endothelial cells(HUVECs) were treated with 10 μmol/L A...Objective To investigate the effects of endothelial microvesicles(EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes. Methods Human umbilical vein endothelial cells(HUVECs) were treated with 10 μmol/L A23187 for 30 min. EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. EMVs were characterized using 1 and 2 μm latex beads and antiPE-CD144 antibody by flow cytometry. For functional research, EMVs at different concentrations were cocultured with H9c2 cardiomyocytes for 6 h. Cell viability of H9c2 cells and the activity of LDH leaked from H9c2 cells were tested by colorimetry. Moreover, apoptosis of H9c2 cells was observed through Hoechst 33258 staining and tested by FITC-Annexin V/PI double staining. Results EMVs were induced by A23187 on HUVECs, and isolated by ultracentrifugation. We identified the membrane vesicles(< 1 μm) induced by A23187 were CD144 positive. In addition, the EMVs could significantly reduce the viability of H9c2 cells, and increase LDH leakage from H9c2 cells in a dose dependent manner(P<0.05). Condensed nuclei could be observed with the increasing concentrations of EMVs through Hoechst 33258 staining. Furthermore, increased apoptosis rates of H9c2 cells could be assessed through FITC-Annexin V/PI double staining by flow cytometry. Conclusion Microvesicles could be released from HUVECs after induced by A23187 through calcium influx, and these EMVs exerted a pro-apoptotic effect on H9c2 cells by induction of apoptosis.展开更多
文摘目的:研究孕酮与钙离子载体A23187诱导顶体反应(Acrosom e reaction,AR)特征。方法:10例健康生育男性精液,采用Percoll密度梯度分离法优选精子,采用FITC-PSA荧光染色法分析AR。结果:A23187诱导AR类型主要为III型和IV型,孕酮诱导AR类型主要为II型。精子获能4、6、8和16 h时,A23187和孕酮诱导AR率与获能0 h组相比较,均明显增加,两组比较差异均有显著性(P<0.05),同一获能时间A 23187和孕酮诱导的AR率比较,差异均无显著性(P>0.05)。A23187诱导15 m in、30 m in和60 m in时,AR率均明显高于诱导前AR率(P<0.005)。孕酮诱导15 m in时,AR率与诱导前组比较没有明显差异(P>0.05),孕酮诱导30 m in和60 m in时,AR率均明显高于诱导前组(P<0.005),同一诱导时间,A23187诱导AR率均明显高于孕酮诱导的AR率,差异均有显著性(P<0.05)。结论:孕酮可诱导精子发生AR,诱导AR类型主要为II型,可比A23187诱导的AR更接近生理过程。
文摘The aim of the study was to prepare berberine hydrochloride long-circulating liposomes and optimize the formulation and process parameters,and investigate the influence of different factors on the encapsulation efficiency.Berberine hydrochloride liposomes were prepared in response to a transmembrane ion gradient that was established by ionophore A23187.Free and liposomal drug were separated by cation exchange resin,and then the amount of intraliposomal berberine hydrochloride was determined by UV spectrophotometry.The optimized encapsulation efficiency of berberine hydrochloride liposomes was 94.3%2.1%when the drug-to-lipid ratio was 1:20,and the mean diameter was 146.9 nm3.2 nm.As a result,the ionophore A23187-mediated ZnSO_(4)gradient method was suitable for the preparation of berberine hydrochloride liposomes that we could get the desired encapsulation efficiency and drug loading.
文摘Effects of procainamide (PA) on human platelet aggregation and the cytosolic free-Ca2+ concentration ([Ca2+]) were investigated in vitro. PA at doses of 8.5 , 34 and 136 μmol·L-1 could inhibit the human platelet aggregation induced by 0. 5 μmol · L-1 A23187 with a good concentration -effect relationship. One min and maximal aggregation rates were also inhibited ( P<0. 01 ,vs control). The [Ca2+], was decreased in the presence of PA ,and the changes of [Ca2+], showed a significant linear correlation with 1 min or maximal aggregation rate (P<0.05). These results suggest that the mechanisms of PA inhibiting platelet aggregation relate to the decrease of [Ca2+].
基金supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China(20101202110005)the Natural Science Foundation of Tianjin(11JCZDJC18300)+1 种基金the Research Foundation of Tianjin Municipal Education Commission(20110106)the National Key Basic Research Program of China(973 Program,2011CB933100)
文摘Objective To investigate the effects of endothelial microvesicles(EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes. Methods Human umbilical vein endothelial cells(HUVECs) were treated with 10 μmol/L A23187 for 30 min. EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. EMVs were characterized using 1 and 2 μm latex beads and antiPE-CD144 antibody by flow cytometry. For functional research, EMVs at different concentrations were cocultured with H9c2 cardiomyocytes for 6 h. Cell viability of H9c2 cells and the activity of LDH leaked from H9c2 cells were tested by colorimetry. Moreover, apoptosis of H9c2 cells was observed through Hoechst 33258 staining and tested by FITC-Annexin V/PI double staining. Results EMVs were induced by A23187 on HUVECs, and isolated by ultracentrifugation. We identified the membrane vesicles(< 1 μm) induced by A23187 were CD144 positive. In addition, the EMVs could significantly reduce the viability of H9c2 cells, and increase LDH leakage from H9c2 cells in a dose dependent manner(P<0.05). Condensed nuclei could be observed with the increasing concentrations of EMVs through Hoechst 33258 staining. Furthermore, increased apoptosis rates of H9c2 cells could be assessed through FITC-Annexin V/PI double staining by flow cytometry. Conclusion Microvesicles could be released from HUVECs after induced by A23187 through calcium influx, and these EMVs exerted a pro-apoptotic effect on H9c2 cells by induction of apoptosis.
