摘要
目的 :探讨甲状腺素对新生大鼠心脏细胞中蛋白激酶C(proteinkinaseC ,PKC)信号途径的影响。 方法 :培养新生大鼠心肌细胞及成纤维细胞 ,用 1%血清培养基或血管紧张素Ⅱ(angiotensinⅡ ,AngⅡ)处理细胞 2 4h后 ,加入甲状腺素(三碘甲状腺素原氨酸 ,triiodothyronine,T3 )继续培养 4 8h后 ,用PKC活性检测试剂盒检测细胞中PKC活性 ,用West ernblot的方法检测细胞中PKCα及PKCε的表达。结果 :在 1%血清培养基中 ,T3 能明显抑制心肌细胞中PKC活性 ,使心肌细胞中PKCε表达下降 ,对PKCα的表达却没有显著的影响 ;在心肌成纤维细胞中 ,无论是PKC活性还是PKCα及PKCε的表达均未观察到T3 的调控作用。预先用AngⅡ处理 2 4h后 ,心肌细胞及心肌成纤维细胞中PKC活性明显增加 ,PKCε的表达显著增加 ,随后用T3 处理后 ,心肌细胞中PKC活性及PKCε的表达明显降低 ;而心肌成纤维细胞中PKC活性没有发生显著性的变化。结论 :甲状腺素能明显抑制心肌细胞中PKC活性及PKCε亚型的表达 ,其对心肌细胞中PKC信号途径的调控作用可能在心肌的多种病理生理过程中起着重要的作用。
Aim:To study the effect of thyroid hormone on protein kinase C activity and isoprotein expressions in cardiac myocytes and fibroblasts of rats in vitro. Methods:Cardiac myocytes and fibroblasts were cultured according to the method of Simpson. Cells were pretreated with 1% newborn calf serum(NCS) or AngiotensinⅡ(AngⅡ) for 24 hours,then Triiodothyronine(T 3) was added to the culture medium and the culture was kept for another 48 hours. The protein kinase C activation were measured by PepTag non-radioactive PKC assay,and the expressions of PKC α and PKC ε were detected by Western blot method. Results:At the condition of 1% NCS culture medium,T 3 could inhibit PKC activity and PKC ε expression in cardiac myocytes significantly,but the expression of PKC α in cardiac myocytes was not influenced by T 3. In cardiac fibroblasts,neither PKC activity nor PKC α and PKC ε expressions was influenced by T 3. When cells were pretreated with AngⅡ for 24 hours,PKC activities in cardiac myocytes and fibroblasts were increased significantly,and PKC ε expressions in cardiac myocytes were also markedly increased. Following a T 3 treatment,PKC activity and PKC ε expression in cardiac myocytes were markedly decreased,but PKC activity in cardiac fibroblasts was not changed. Conclusion:Whether at the condition of 1% NCS medium or in a pretreatment with AngⅡ,thyroid hormone could inhibit the PKC activity and PKC ε expression in cardiac myocytes. The influence of thyroid hormone on the PKC signal pathway in cardiac myocyte may be involved in many pathophysiological progress of myocardium.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2004年第1期38-41,共4页
Chinese Journal of Applied Physiology
基金
湖北省自然科学基金 (980 91)
