AIM: To purify and identify heme oxygenase (HO) isomers which exist in rat liver, spleen and brain treated with hematin and phenylhydrazine and in untreated rat liver and to investigate the characteristics of HO isome...AIM: To purify and identify heme oxygenase (HO) isomers which exist in rat liver, spleen and brain treated with hematin and phenylhydrazine and in untreated rat liver and to investigate the characteristics of HO isomers, to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey kidney cells (COS-1 cells), to prepare the rat heme oxygenase-1 (HO-1) mutant and to detect inhibition of HO-1 mutated enzyme.METHODS: First, rat liver, spleen and brain microsomal fi-actions were purified by DEAE-Sephacel and hydroxylapatite. The characteristics including activity, immunity and inducibility of two isomers (HO-1 and HO-2), and their apparent molecular weight were measured by detecting enzymatic activities, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis, respectively. Second, plasmid pcDNA3HO1 containing native rat HO-1 cDNA and pcDNA3HO1D25 carrying mutated rat HO-1 cDNA (His25Ala) were constructed by site-directed mutagenesis. COS-1 cells transfected with pcDNA3HO1 and pcDNA3HO1D25 were collected and disrupted by sonication, the microsomes were prepared by ultracentrifugation. Third, the inhibition of rat HO-1 mutant was analyzed.RESULTS: Two isomers were purified and identified in treated rat liver, spleen, brain and untreated rat liver. HO-1 was the predominant form with a ratio of 2.0:1 and 3.2:1 of HO-1 and HO-2 in liver and spleen, respectively, but only the activity of HO-2 in the brain and untreated liver could be detected. The apparent molecular weights of HO-1 and HO-2 were about Mr 30 000 and Mr 36 000 under reducing conditions, respectively. The antiserum against liver HO-2 was employed in Western blotting analysis, the reactivity of HO-1 in the liver was not observed. The plasmid pcDNA3HO1 was highly expressed in endoplasmic reticulum of transfected COS-1 cells. The specific activity was ≈5-fold higher than that of the control. However, the enzyme activity of mutated HO-1 declined. While an equal amount of mutant was added to the enzyme reaction system, the levels of bilirubin decreased 42 %.CONCLUSION: The studies suggest that HO-1 and HO-2 exist in the hematin and phenylhydrazine treated rat liver and spleen, but only HO-2 in the brain and untreated liver.Two constitutive forms are different in molecular weight, inducibility and immunochemical properties. The activity of expressed HO-1 in COS-1 cells is higher than that of purified enzyme from rat spleen tissue. It suggests that this clone has an insert of 1030 base-pairs encodes HO-1. His25Ala mutant reduced the formation of bilirubin and it suggests that the mutant could competely bind the heme with native enzyme.展开更多
Objective:To study the pharmacological effect of Xiaoke Granule (XKG, 消渴冲剂) on blood sugar and blood rheological property in the diabetic animals.Methods: Alloxan induced diabetic mice or rats were grouped randoml...Objective:To study the pharmacological effect of Xiaoke Granule (XKG, 消渴冲剂) on blood sugar and blood rheological property in the diabetic animals.Methods: Alloxan induced diabetic mice or rats were grouped randomly. The effects of XKG on blood sugar, appetite, capacity of drinking, glucose tolerance, blood lipid and blood rheological property were observed and compared among groups.Results:XKG showed a trend in reducing the appetite and capacity of drinking, increasing the body weight, and significantly inhibiting the increase of blood sugar coused by ectogenic glucose in mice, and could improve the blood lipid and blood rheological property in rats.Conclusion:XKG is effective in reducing serum total cholesterol, lowering the blood viscosity, improving the blood rheological property of alloxan induced diabetic animals. Therefore, it might effective in treating and preventing the occurrence of diabetes mellitus and the complications of blood stasis.展开更多
每年，世界顶级的大师和品牌都会带着他们的最新作品来到MAISON＆OBJET，为我们搭建起美轮美奂的舞台，共同庆祝最新创意的诞生。每一届展会上，都有新的艺术家获得专业人士的肯定并被荣幸地评为各个主题展的年度设计师。当然，巴黎家...每年，世界顶级的大师和品牌都会带着他们的最新作品来到MAISON＆OBJET，为我们搭建起美轮美奂的舞台，共同庆祝最新创意的诞生。每一届展会上，都有新的艺术家获得专业人士的肯定并被荣幸地评为各个主题展的年度设计师。当然，巴黎家居装饰博览会也无可比拟地成为了年轻人才们跨越成功的跳板。不论是Scènes d’intérieur’s主题展上的“Talents à la Carte”亦或是Atelier d’Artde France的青年设计师竞赛都为明日之星的诞生提供最佳平台。展开更多
基金National Natural Scientific Foundation of China,No.39600159
文摘AIM: To purify and identify heme oxygenase (HO) isomers which exist in rat liver, spleen and brain treated with hematin and phenylhydrazine and in untreated rat liver and to investigate the characteristics of HO isomers, to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey kidney cells (COS-1 cells), to prepare the rat heme oxygenase-1 (HO-1) mutant and to detect inhibition of HO-1 mutated enzyme.METHODS: First, rat liver, spleen and brain microsomal fi-actions were purified by DEAE-Sephacel and hydroxylapatite. The characteristics including activity, immunity and inducibility of two isomers (HO-1 and HO-2), and their apparent molecular weight were measured by detecting enzymatic activities, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis, respectively. Second, plasmid pcDNA3HO1 containing native rat HO-1 cDNA and pcDNA3HO1D25 carrying mutated rat HO-1 cDNA (His25Ala) were constructed by site-directed mutagenesis. COS-1 cells transfected with pcDNA3HO1 and pcDNA3HO1D25 were collected and disrupted by sonication, the microsomes were prepared by ultracentrifugation. Third, the inhibition of rat HO-1 mutant was analyzed.RESULTS: Two isomers were purified and identified in treated rat liver, spleen, brain and untreated rat liver. HO-1 was the predominant form with a ratio of 2.0:1 and 3.2:1 of HO-1 and HO-2 in liver and spleen, respectively, but only the activity of HO-2 in the brain and untreated liver could be detected. The apparent molecular weights of HO-1 and HO-2 were about Mr 30 000 and Mr 36 000 under reducing conditions, respectively. The antiserum against liver HO-2 was employed in Western blotting analysis, the reactivity of HO-1 in the liver was not observed. The plasmid pcDNA3HO1 was highly expressed in endoplasmic reticulum of transfected COS-1 cells. The specific activity was ≈5-fold higher than that of the control. However, the enzyme activity of mutated HO-1 declined. While an equal amount of mutant was added to the enzyme reaction system, the levels of bilirubin decreased 42 %.CONCLUSION: The studies suggest that HO-1 and HO-2 exist in the hematin and phenylhydrazine treated rat liver and spleen, but only HO-2 in the brain and untreated liver.Two constitutive forms are different in molecular weight, inducibility and immunochemical properties. The activity of expressed HO-1 in COS-1 cells is higher than that of purified enzyme from rat spleen tissue. It suggests that this clone has an insert of 1030 base-pairs encodes HO-1. His25Ala mutant reduced the formation of bilirubin and it suggests that the mutant could competely bind the heme with native enzyme.
文摘Objective:To study the pharmacological effect of Xiaoke Granule (XKG, 消渴冲剂) on blood sugar and blood rheological property in the diabetic animals.Methods: Alloxan induced diabetic mice or rats were grouped randomly. The effects of XKG on blood sugar, appetite, capacity of drinking, glucose tolerance, blood lipid and blood rheological property were observed and compared among groups.Results:XKG showed a trend in reducing the appetite and capacity of drinking, increasing the body weight, and significantly inhibiting the increase of blood sugar coused by ectogenic glucose in mice, and could improve the blood lipid and blood rheological property in rats.Conclusion:XKG is effective in reducing serum total cholesterol, lowering the blood viscosity, improving the blood rheological property of alloxan induced diabetic animals. Therefore, it might effective in treating and preventing the occurrence of diabetes mellitus and the complications of blood stasis.
文摘每年，世界顶级的大师和品牌都会带着他们的最新作品来到MAISON＆OBJET，为我们搭建起美轮美奂的舞台，共同庆祝最新创意的诞生。每一届展会上，都有新的艺术家获得专业人士的肯定并被荣幸地评为各个主题展的年度设计师。当然，巴黎家居装饰博览会也无可比拟地成为了年轻人才们跨越成功的跳板。不论是Scènes d’intérieur’s主题展上的“Talents à la Carte”亦或是Atelier d’Artde France的青年设计师竞赛都为明日之星的诞生提供最佳平台。
