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八肽胆囊收缩素激活钙离子通道和酪氨酸激酶诱导豚鼠心肌细胞内的游离钙增高

赵晓云, 凌亦凌, 尚忠林, 李清, 尹京湘, 檀国军

河北省老年医学重点实验室.河北,石家庄 050051；河北医科大学病理生理教研室.河北,石家庄 050017；河北师范大学生命科学学院.河北,石家庄 050017；河北医科大学生理教研室.河北,石家庄 050017

摘要

探讨八肽胆囊收缩素(CCK-8)对豚鼠单个心肌细胞内游离钙浓度([Ca~(2+)]_(i))的影响及其信号转导机制。Fluo 3-AM标记酶消化法分离的单个心室肌细胞,用激光共聚焦显微镜测定细胞内[Ca~(2+)]_(i)的浓度。[Ca~(2+)]_(i)的变化用荧光强度(F_(i))和相对荧光强度(F_(i)/F_(0)%)表示。实验结果如下:(1)在含Ca~(2+) 1.0mmol/L的Tyrode^s液中,CCK-8 (1--10~(4) pmol/L)均可引起[Ca~(2+)]_(i)快速显著上升(P<0.01)。(2)用钙离子螯合剂EGTA(3mmol/L)和钙离子通道阻断剂nisoldipine (0.5#mu#mol/L)预孵育心肌细胞5min,CCK-8(10~(2) pmol/L)仅可引起[Ca~(2+)]_(i)缓慢轻度上升(P<0.01)。(3)用非选择性CCK受体拮抗剂丙谷胺(proglumide 6 #mu#mol/L)或酪氨酸激酶抑制剂genistein(1 #mu#mol/L)预孵育心肌细胞5min,则完全抑制CCK-8诱导的[Ca~(2+)]_(i)升高(P<0.01)。CCK-8可通过激活其受体控制的Ca~(2+)通道,引起Ca~(2+)内流,诱导细胞内Ca~(2+)释放,引起豚鼠单个心肌细胞内[Ca~(2+)]_(i)上升,此作用可能由酪氨酸激酶介导。

关键词： cholecystokinin; cardiomyocyte; fluorescent intensity; Ca2+ channel; confocal microscopy; receptor

Cholecystokinin octapeptide increases free intracellular calcium of guinea pig cardiomyocytes through activation of Ca~(2+) channel and tyrosine kinase

Zhao Xiaoyun, Ling Yiling, Shang Zhonglin, Li Qing, Yin Jingxiang, Tan Guojun

Hebei Provincial Geriatric Key Laboratory.Shijiazhuang 050051,Hebei；China；College of Biological Science,Hebei Normal University.Shijiazhuang 050017,Hebei

Abstract

The results obtained are as follows. (1) In the normal Tyrode^s solution containing 1.0 mmol/L Ca~(2+), CCK-8 (1--10~(4) pmol/L) eicited a rapid and marked increase in [Ca~(2+)]_(i)). (2) When cardiomyocytes were pretreated with the Ca~(2+) chelator EGTA (3 mmol/L) and Ca~(2+) channel antagonist nisoldipine (0.5 #mu#mol/L) for 5 min, CCK-8 (10~(2) pmol/L) caused a slow and small increase in [Ca~(2+)]_(i) (P<0.01). (3) Pretreatment with the nonselected CCK-receptor (CCK-R) antagonist proglumide (6 #mu#mol/L) or the tyrosine kinase inhibitor genistein (1 #mu#mol/L) for 5 min could inhibit the increase of [Ca~(2+)]_(i) induced by CCK-8 (10~(2) pmol/L) (P<0.01). The results suggest that CCK-8 increases the [Ca~(2+)]_(i) via activating the receptor-operated Ca~(2+) chanel and eliciting the influx of Ca~(2+) in isolated guinea pig cardiomyocytes, in which tyrosine kinase may be involved.

Key words: 八肽胆囊收缩素;心肌细胞;荧光强度;钙通道;共聚焦显微镜;受体

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引用本文：

赵晓云, 凌亦凌, 尚忠林, 李清, 尹京湘, 檀国军. 八肽胆囊收缩素激活钙离子通道和酪氨酸激酶诱导豚鼠心肌细胞内的游离钙增高[J]. 生理学报 2004; 56 (1): .

Zhao Xiaoyun, Ling Yiling, Shang Zhonglin, Li Qing, Yin Jingxiang, Tan Guojun. Cholecystokinin octapeptide increases free intracellular calcium of guinea pig cardiomyocytes through activation of Ca~(2+) channel and tyrosine kinase. Acta Physiol Sin 2004; 56 (1): (in Chinese with English abstract).