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.
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2015年第1期40-45,共6页
Acta Laboratorium Animalis Scientia Sinica
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)
关键词
脾气虚证
钙调蛋白
蛋白
肝组织
大鼠
Spleen-qi deficiency
CaM, CaMK II, p-CaMK II
Protein
Liver
Rats
