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脾气虚大鼠肝组织三磷酸腺苷酶活性和CaMKⅡ通路CaM、CaMKⅡ蛋白表达的变化     被引量:3

Changes of adenosine triphosphatase activity and expression of CaM and CaMK Ⅱ protein in CaMK Ⅱ signaling pathways in the liver tissue of rats with spleen-qi deficiency

文献类型:期刊文献

中文题名:脾气虚大鼠肝组织三磷酸腺苷酶活性和CaMKⅡ通路CaM、CaMKⅡ蛋白表达的变化

英文题名:Changes of adenosine triphosphatase activity and expression of CaM and CaMK Ⅱ protein in CaMK Ⅱ signaling pathways in the liver tissue of rats with spleen-qi deficiency

作者:成映霞[1];段永强[1,2];梁玉杰[1,3];杨晓轶[1,3];杜娟[1];刘靓[1];程卫东[2];朱立鸣[1];安耀荣[1];高建德[1]

第一作者:成映霞

机构:[1]甘肃中医学院、中医方药创新工程重点实验室,兰州730020;[2]兰州大学基础医学院中西医结合研究所,兰州730020;[3]甘肃中医学院、甘肃省中药药理与毒理学重点实验室,兰州730020

第一机构:甘肃中医药大学

年份:2015

卷号:23

期号:1

起止页码:40

中文期刊名:中国实验动物学报

外文期刊名:Acta Laboratorium Animalis Scientia Sinica

收录:CSTPCD;;北大核心:【北大核心2014】;CSCD:【CSCD_E2015_2016】;

基金:国家自然科学基金(81160420);甘肃省自然科学基金(1010RJZA148);甘肃省教育厅基金(1006-05)

语种:中文

中文关键词:脾气虚证;钙调蛋白;蛋白;肝组织;大鼠

外文关键词:Spleen-qi deficiency; CaM, CaMK II, p-CaMK II; Protein; Liver; Rats

摘要:目的观察脾气虚大鼠肝组织三磷酸腺苷酶活性和CaMKⅡ通路关键分子[Ca2+]i以及CaM、CaMKⅡ、p-Ca MKⅡ蛋白表达水平的变化。方法受试动物随机分为4组,即空白对照组,脾气虚模型7 d、14 d、21d组,每组12只。除空白对照组外,其余受试动物采用复合法(苦寒破气法、游泳力竭法及饥饱失常法)成功建立脾气虚证大鼠模型,在观测各组大鼠一般生存状态和肝组织ATP酶活性的基础上,采用激光共聚焦技术检测肝组织细胞内[Ca2+]i浓度,蛋白免疫印迹技术检测肝组织CaM、CaMKⅡ和p-CaMKⅡ的表达变化。结果与空白组比较,脾气虚大鼠随着造模时间的延长,一般生存状况渐差,肝组织Na+-K+-ATPase和Ca2+-Mg2+-ATPase活性均降低(P<0.05,P<0.01);肝组织[Ca2+]i浓度和CaM、CaMKⅡ、p-Ca MK Ⅱ蛋白表达量显著降低(P<0.01);且脾气虚模型7 d、14 d、21 d组之间比较,以脾气虚21 d组变化更为显著。结论脾气虚大鼠肝组织Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性和[Ca2+]i浓度降低,并且CaMKⅡ信号通路关键分子CaM、CaMKⅡ和p-CaMKⅡ蛋白表现为低表达。
Objective To investigate the changes of adenosine triphosphatase activity, expression of cahnodulin (CaM) , and ealmodulin-dependent protein kinase II (CaMK II) protein in CaMK II signaling pathways in the liver tissue of rats with spleen-qi deficiency. Methods 3-month-old SPF Wistar rats ( male : female = 1 : 1 ) were randomly divided in- to 4 groups : normal control group, and three spleen-qi deficient model groups ( observed on days 7, 14 and 21 ) , 12 rats in each group. The spleen-qi deficient rat models were generated by bitter-cold purgatives, exhaustion and fully or poorly feed- ing food in turn, and their general conditions and ATPase activity were evaluated. Confocal laser scanning microscopy was used to detect cellular [ Ca^2+ ] i concentrations and Western blotting was used to test CaM, CaMK II, p-CaMK II expression in liver tissue of the spleen-qi deficient rats. Results Compared with the normal group, general condition was poor, activ- ities of Na^+ -K^+ -ATPase and Ca^2 + -Mg^2 + -ATPase were significantly decreased ( P 〈 0.01 , P 〈 0. 05 ) , and expressions of CaM, CaMK II, and p-CaMK II in the liver tissue were decreased. A most obvious difference was observed in the spleen- qi deficient model day 21 group ( P 〈 0.01 ). Conclusions Our results demonstrate that decease of adenosine triphos- phatase activities and [ Ca^2+ ] i concentration is involved in the spleen-qi deficiency, and expressions of key molecules of CaMK II signaling pathway, CaM, CaMK Ⅲ and p-CaMK II proteins, are also decreased.

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