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目的观察离体活膀胱肌条铺片中卡哈尔间质细胞(interstitial cells of cajal,ICCs)自发性钙瞬变,并检测胞外钙及胞内钙库钙对ICCs细胞自发性钙瞬变的影响,探讨ICCs细胞自发性钙瞬变产生的机制。方法制作离体膀胱肌条铺片,生理液灌流保持肌条铺片活性。待肌条自发性收缩活动出现后,Fluo-4负载肌条铺片,激光共聚焦显微镜观察其中ICCs细胞自发性钙瞬变,随后观察尼莫地平(Nimodipine)、2-氨基乙基二苯硼酸盐(2-aminoethoxydip henylborate,2-APB)、雷诺丁(Ryanodine)、毒胡萝卜素(Thapsigargin)及无钙、高钙生理液对ICCs细胞自发性钙瞬变的影响,探讨ICCs细胞自发性钙瞬变产生的机制。结果可见离体活膀胱肌条铺片中平滑肌细胞和ICCs细胞均产生节律性自发性钙瞬变,平滑肌细胞钙瞬变频率较快,而ICCs细胞钙瞬变频率相对较慢。尼莫地平能完全抑制平滑肌细胞的自发性钙瞬变,但对ICCs细胞的自发性钙瞬变无显著影响。2-APB、雷诺丁、毒胡萝卜素及无钙生理液均可完全抑制ICCs细胞的自发性钙瞬变,高钙生理液可显著增加ICCs细胞自发性钙瞬变的频率。结论细胞外钙离子通过非L型钙离子通道入胞及细胞内钙库放钙均参与ICCs细胞自发性钙瞬变的产生。
Objective To observe the spontaneous calcium transients in interstitial cells of cajal (ICCs) in vitro and to detect the effects of extracellular calcium and intracellular Ca2 + on the spontaneous calcium transients To investigate the mechanism of spontaneous calcium transients in ICCs. Methods Preparation of bladder muscle strip in vitro, physiological fluid perfusion to keep the activity of muscle strips. After the spontaneous contractions of myoblasts appeared, the spontaneous calcium transients of ICCs were observed by confocal laser scanning microscopy with Fluo-4 loaded strips, followed by the observation of Nimodipine, 2-aminoethyldiphenylboron 2-APB, Ryanodine, Thapsigargin and calcium-free and calcium-rich physiological fluids on the spontaneous calcium transients in ICCs were investigated. The effects of 2-aminoethoxydiphenyborate (2-APB), Ryanodine, Mechanism of transient generation. The results showed that the spontaneous Ca2 + transient changes in rhythmic spontaneous calcium transients were found in both smooth muscle cells and ICCs in vitro, but the frequency of calcium transients in smooth muscle cells was faster, while the frequency of calcium transients in ICCs was relatively slow. Nimodipine completely inhibited spontaneous calcium transients in smooth muscle cells but had no significant effect on spontaneous calcium transients in ICCs. 2-APB, Raynaudine, thapsigargin and calcium-free physiological fluids could completely inhibit the spontaneous calcium transients of ICCs cells. The high calcium physiological fluids could significantly increase the frequency of spontaneous calcium transients in ICCs. CONCLUSION: Extracellular calcium ions are involved in the spontaneous Ca2 + transient translocation in ICCs through non-L-type Ca2 + channel entry and intracellular Ca2 + uptake.