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目的观察大鼠离体活膀胱肌条铺片中平滑肌细胞自发性钙瞬变,并探讨平滑肌细胞自发性钙瞬变产生的机制。方法制作大鼠离体膀胱肌条铺片,生理液灌流保持肌条铺片活性。采用Fluo-4负载肌条铺片,激光共聚焦显微镜法观察其中平滑肌细胞自发性钙瞬变,并观察尼莫地平(Nimodipine)、2-氨基乙基二苯硼酸盐(2-aminoethoxydiphenylbo-rate,2-APB)、雷诺丁(Ryanodine)、毒胡萝卜素(Thapsigargin)对平滑肌细胞自发性钙瞬变的影响,探讨平滑肌细胞自发性钙瞬变产生的机制。结果可见大鼠离体活膀胱肌条铺片中平滑肌细胞产生节律性自发性钙瞬变,自发性钙瞬变幅度(F/F0)和频率分别为(2.20±0.13)和(10.31±2.74)次/min。L型钙离子通道阻断剂Nimodipine能完全抑制平滑肌细胞的自发性钙瞬变,2-APB、Ryanodine可将钙瞬变幅度分别降低至(1.98±0.14)、(1.81±0.11),而相应的钙瞬变频率则被减少为(8.18±1.97)(P<0.05,n=18)、(7.59±2.16)次/min(P<0.01,n=17)。Thapsigargin可抑制平滑肌自发性钙瞬变幅度至(1.76±0.11)(P<0.01,n=20),但对钙瞬变频率却无显著影响。结论细胞外钙离子通过L型钙离子通道入胞在平滑肌细胞自发性钙瞬变产生中起主要作用,而细胞内钙库中的钙释放入细胞质可能起辅助作用。
Objective To observe the spontaneous calcium translocation of smooth muscle cells in vitro and to explore the mechanism of spontaneous calcium translocation in smooth muscle cells. Methods The isolated bladder myoblasts of rats were made, and the physiological fluid was infused to keep the activity of muscle strips. The spontaneous calcium transients of smooth muscle cells were observed by laser scanning confocal microscopy with Fluo-4 loaded muscle strips, and the effects of Nimodipine, 2-aminoethoxydiphenylbo-rate , 2-APB, Ryanodine and Thapsigargin on spontaneous calcium transients of smooth muscle cells, and to explore the mechanism of spontaneous calcium transients in smooth muscle cells. The results showed that the spontaneous Ca2 + transient amplitude (F / F0) and the frequency of spontaneous Ca2 + translocation were (2.20 ± 0.13) and (10.31 ± 2.74), respectively, in smooth muscle cells of rat bladder myocytes. Times / min. Lim-type calcium channel blocker Nimodipine can completely inhibit the spontaneous calcium transients of smooth muscle cells. The 2-APB and Ryanodine can reduce the amplitude of calcium transient to (1.98 ± 0.14) and (1.81 ± 0.11) respectively, while the corresponding Calcium transient frequency was reduced to (8.18 ± 1.97) (P <0.05, n = 18), (7.59 ± 2.16) times / min (P <0.01, n = 17). Thapsigargin inhibited smooth muscle spontaneous calcium transient amplitude (1.76 ± 0.11) (P <0.01, n = 20), but had no significant effect on the frequency of calcium transients. Conclusion Extracellular calcium plays a major role in the spontaneous Ca2 + transient translocation via L-type Ca2 + channel, and Ca2 + release into the cytoplasm may play an auxiliary role.