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目的 通过对家兔阴茎海绵体平滑肌细胞 (PCSMC)内游离Ca2 + 检测方法的对比 ,以选择更适宜的方法来研究阴茎海绵体平滑肌的舒张机制。方法 采用新型Ca2 + 荧光探针Fluo 3/AM标记家兔PCSMC胞浆内游离Ca2 + ,应用显微分光光度计 (MSP)、流式细胞仪 (FCM)和激光扫描共聚焦显微镜 (LSCM)分别观察硝苯吡啶 (NIF)对高K+ 诱发家兔PCSMC内 [Ca2 + ]i 变化的影响。结果 MSP和FCM检测发现 ,10 μmol/LNIF能明显减弱高K+ (4 0mmol/L)诱发细胞内荧光强度的升高 ,抑制率为 (6 5 .5± 4 .4 ) %和 (6 9.3± 5 .2 ) %。LSCM可见细胞胞浆内游离Ca2 +分布 ,不同细胞胞浆内的基础荧光强度有差别 ,NIF能明显抑制高K+ 诱发的细胞内 [Ca2 + ]i 升高 ,抑制率为 (71.5±6 .8) %。结论 LSCM能直观地看到单细胞内Ca2 + 的分布 ,并能动态观察胞浆内 [Ca2 + ]i 的变化 ,为研究不同药物对PCSMC的影响提供了良好的实验方法。
Objective To compare the detection of free Ca2 + in rabbit corpus cavernosum smooth muscle cells (PCSMC) and select the more appropriate method to study the relaxation mechanism of the smooth muscle of the penis. Methods Free Ca2 + in cytoplasm of rabbit PCSMC was labeled with Fluo 3 / AM, a new Ca2 + fluorescent probe. The expression of free Ca2 + in rabbit cytoplasm was detected by using MSP, FCM and LSCM respectively. To observe the effect of nifedipine (NIF) on [Ca2 +] i in high-K + -induced rabbit PCSMC. Results MSP and FCM showed that 10 μmol / L LNIF could significantly attenuate the increase of intracellular fluorescence intensity induced by high K + (40 mmol / L), with the inhibition rates of (6.55 ± 4. 4)% and (6 9.3 ± 5 .2)%. LSCM showed intracellular free Ca2 + distribution in the cytoplasm, the difference between the basic fluorescence intensity in different cytoplasm, NIF can significantly inhibit high K + -induced intracellular [Ca2 +] i increased, the inhibition rate was (71.5 ± 6.8 )%. Conclusion LSCM can intuitively observe the distribution of intracellular Ca2 +, and can dynamically observe the changes of [Ca2 +] i in the cytoplasm. It provides a good experimental method for studying the influence of different drugs on PCSMC.