【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。更多还原

【Abstract】 Objective To investigate the changes of adenosine triphosphatase activity,expression of calmodulin( Ca M),and calmodulin-dependent protein kinase Ⅱ( Ca MK Ⅱ) protein in Ca MK 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 into 4 groups: normal control group,and three spleen-qi deficient model groups( observed on days 7,14 and 21),12 rats ineach group. The spleen-qi deficient rat models were generated by bitter-cold purgatives,exhaustion and fully or poorly feeding food in turn,and their general conditions and ATPase activity were evaluated. Confocal laser scanning microscopy was used to detect cellular [Ca2 +]i concentrations and Western blotting was used to test CaM,CaM K Ⅱ,p-CaM K Ⅱ expression in liver tissue of the spleen-qi deficient rats. Results Compared with the normal group,general condition was poor,activities of Na+-K+-ATPase and Ca2 +-Mg2 +-ATPase were significantly decreased( P < 0. 01,P < 0. 05),and expressions of CaM,CaM K Ⅱ,and p-CaM K Ⅱ in the liver tissue were decreased. A most obvious difference was observed in the spleenqi deficient model day 21 group( P < 0. 01). Conclusions Our results demonstrate that decease of adenosine triphosphatase activities and [Ca2 +]i concentration is involved in the spleen-qi deficiency,and expressions of key molecules of CaM K Ⅱ signaling pathway,CaM,CaM K Ⅱ and p-CaM K Ⅱ proteins,are also decreased.更多还原