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目的:研究酪氨酸激酶是否参与α_(1A)肾上腺素受体介导的人胚胎肾细胞(HEK293)浆游离钙浓度调节。方法:用Fura-2/AM荧光法测定细胞游离钙浓度,底物反应法测定酪氨酸激酶活性。结果:去甲肾上腺素(NE)可引起的HEK293细胞内游离钙浓度升高至(371±31)nmol·L~(-1),酪氨酸激酶抑制剂檞皮素和tyrphostin可抑制NE引起的细胞内游离钙浓度升高,但不抑制NE引起磷酸肌醇的生成,NE可引起HEK293细胞浆酪氨酸激酶升高1.73±0.72倍,这种作用可被PKC的抑制剂calphostin C和细胞内钙耗竭所抑制。结论:酪氨酸激酶参与α_1A肾上腺素受体介导的人胚胎肾细胞内游离钙浓度升高。
AIM: To investigate whether tyrosine kinase is involved in the regulation of free calcium concentration in α_ (1A) adrenergic receptor-mediated human embryonic kidney cell line (HEK293). Methods: Fura-2 / AM fluorescence method was used to determine the cell free calcium concentration, and substrate reaction method was used to determine tyrosine kinase activity. Results: The intracellular free calcium concentration in HEK293 cells induced by norepinephrine (NE) increased to (371 ± 31) nmol·L -1, while the tyrosine kinase inhibitors nobiletin and tyrphostin inhibited the NE-induced Of intracellular free calcium concentration, but did not inhibit NE-induced phosphoinositide production. NE induced a 1.73 ± 0.72-fold increase of tyrosine kinase in HEK293 cells. This effect was enhanced by PKC inhibitor calphostin C and cells Inhibition of calcium depletion. CONCLUSION: Tyrosine kinase is involved in the elevation of intracellular free calcium concentration in human embryonic kidney cells mediated by α_1A adrenergic receptor.