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目的观察不同浓度的丙泊酚对体外培养的兔气管黏膜上皮细胞纤毛运动的影响,探讨一氧化氮(NO)在丙泊酚影响纤毛运动中的作用。方法无菌条件下从健康新西兰兔获取离体气管标本,分离上皮组织培养7～8d后观察。以Hank平衡盐溶液(HBSS)将丙泊酚的终浓度分别稀释为1、10、20、50、100、200μg/ml。将组织块随机分为8组,前7组分别加入HBSS(P0)和上述6种浓度的丙泊酚(P1、P2、P3、P4、P5、P6),第8组(P7)在加入0.1mmol/L的一氧化氮合酶(NOS)总抑制剂——L-NMMA后再以200μg/ml丙泊酚处理。在加药前(T0),加药后1min(T1)、5min(T2)、10min(T3)、20min(T4)、30min(T5),采用高速数码摄像系统采集纤毛摆动图像,通过IPLabV3.65软件分析,计算纤毛摆动频率(ciliarybeatfrequency,CBF),分别进行组内和组间对比。结果①P0组、P1组和P2组的CBF值在各时点与各自的基础值比较均无明显变化;P1组和P2组的CBF值与P0组相应时间点相比无明显变化;②P3组CBF在T1时点比基础值和P0组T1时点值明显升高(P<0.05);在随后的各个观察时点与基础值及P0组相应时间点比较无明显变化;③P4、P5、P6组加药后各个观察时点的CBF均明显高于基础值和P0组各时点值(P<0.05);④P7组加药后各时点的CBF值与基础值相比均无明显变化。结论50μg/ml以上浓度的丙泊酚可促进纤毛摆动,50、100、200μg/ml浓度的丙泊酚可能通过促进NO合成和释放而加速离体纤毛运动。
Objective To observe the effects of different concentrations of propofol on the ciliary motility of rabbit tracheal epithelial cells cultured in vitro and to explore the role of nitric oxide (NO) in the ciliary motility induced by propofol. Methods Exsemporal tracheal samples were obtained from healthy New Zealand rabbits under aseptic conditions. The epithelial tissues were isolated and cultured for 7-8 days. The final concentrations of propofol were respectively diluted to 1, 10, 20, 50, 100, 200 μg / ml with Hank’s Balanced Salt Solution (HBSS). Tissue blocks were randomly divided into 8 groups: HBSS (P0) and propofol (P1, P2, P3, P4, P5 and P6) mmol / L of nitric oxide synthase (NOS) total inhibitor - L-NMMA followed by 200 μg / ml propofol. The ciliary wobble images were collected by high-speed digital camera system before injection (T0), 1min (T1), 5min (T2), 10min (T3), 20min (T4) and 30min (T5) Software analysis, calculation of ciliarybeatfrequency (CBF) frequency, intra-group and inter-group comparison. Results ① There was no significant difference in CBF value between P0 group, P1 group and P2 group at each time point with their respective baseline values. There was no significant difference between CBF value of P1 group and P2 group at the corresponding time point of P0 group. ②P3 (P <0.05), and there was no significant difference between baseline value and the corresponding time points in P0 group at all time points after the observation; ③P4, P5, P6 group The CBF of each observation point after dosing were significantly higher than the baseline value and the time points of P0 group (P <0.05) .④The CBF value of each group after P7 administration did not change significantly compared with the baseline values. Conclusion Propofol at concentrations above 50 μg / ml promotes ciliary wobble. Propofol at concentrations of 50, 100 and 200 μg / ml may accelerate isolated ciliary motility by promoting NO synthesis and release.