摘要
目的 探讨M3受体激动对H2 O2 诱导的大鼠培养心肌细胞凋亡的作用 ,进一步阐明其机制。方法 末端标记法 (TUNEL)进行细胞凋亡检测 ;免疫组化方法检测Bcl 2和Fas的表达 ;共聚焦显微镜观察 [Ca2 + ]i荧光强度变化。结果 M3受体激动剂胆碱 (10mmol·L- 1 )可减少H2 O2 诱导的心肌细胞凋亡的数量 ,并可增加心肌Bcl 2的表达 ,减少Fas表达 ,抑制H2 O2 诱导的 [Ca2 + ]i荧光强度的升高。但预先应用 4DAMP (10nmol·L- 1 )阻断M3受体可逆转胆碱作用。结论 激动M3受体对H2 O2 诱导的心肌细胞凋亡有保护作用 ,其机制可能与Bcl 2和Fas表达以及下调[Ca2 + ]i有关。
Aim To observe the effect of activation of M 3 receptor on H 2O 2 induced apoptosis in cultured rat myocytes and to investigate its possible mechanisms. Methods Isolated neonatal cardiomyocytes were cultured. Morphologic changes were observed by microscopy. The apoptosis in cardiomyocyte was detected by terminal deoxynucleotide transferase directed d-UTP nick and end labeling (TUNEL) assay. The expression of apoptosis-related protein in Bcl-2 and Fas was measured by immunohistochemistry assay. [Ca 2+] i in single cardiomyocyte loaded with Fluo 3-AM was measured by confocal microscope. Results H 2O 2-mediated myocyte apoptosis was attenuated by M 3 receptor agonist choline (10 mmol·L -1). Pretreatment of cardiac myocytes with choline also increased Bcl-2, decreased Fas expression, and inhibited the increase in FI value of [Ca 2+] i in H 2O 2-stimulated cardiac myocytes. However, blockade of M 3 receptor by 4DAMP (10 nmol·L -1) completely inhibited the effects of choline on H 2O 2-stimulated cardiac myocytes. Conclusion Activation of M 3 receptor showed protective effect on H 2O 2-induced apoptosis in cultured rat myocytes and this effect might be related to modulating the expression of some genes including Bcl-2 and Fas as well as the downregulation of [Ca 2+] i.
出处
《药学学报》
CAS
CSCD
北大核心
2004年第11期887-891,共5页
Acta Pharmaceutica Sinica
基金
NationalScienceFundationofChina (3 0 2 715 99) .
