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目的:探讨H_2O_2对幼年斑马鱼侧线毛细胞发育及损伤后再生的影响。方法:选取受精后5d(5dpf)斑马鱼,随机分为对照组:系统水培养;H_2O_2组:10μmol/L、20μmol/L H_2O_2溶液培养;新霉素组:200μmol/L硫酸新霉素处理1h后用系统水培养;新霉素+H_2O_2组:200μmol/L硫酸新霉素处理1h后用20μmol/L H_2O_2溶液培养;顺铂组:1 000μmol/L顺铂处理3h后用系统水培养;顺铂+H_2O_2组:1 000μmol/L顺铂处理3h后用20μmol/L H_2O_2溶液培养。各组在H_2O_2处理0、24、48h时用免疫荧光法标记侧线毛细胞并在荧光显微镜下选取P1、P7、P8三个神经丘计数。实验重复3次。结果:5~7dpf斑马鱼P1、P7、P8三个神经丘毛细胞数量分别为(9.364±0.901,n=11),(9.645±0.598,n=15)和(9.922±0.862,n=13),两两之间无明显差异(P>0.05);10μmol/L、20μmol/L H_2O_2处理5dpf斑马鱼48h后,侧线毛细胞数量分别为11.540±0.741、11.905±0.607,与对照组(10.841±0.389)比较差异均有统计学意义(P<0.05);新霉素+H_2O_248h组与新霉素48h组毛细胞数分别为10.600±0.689、8.767±0.603,2组比较差异有统计学意义(P<0.01);顺铂+H_2O_248h组与顺铂48h组毛细胞数分别为5.967±1.086、5.633±1.548,2组比较差异无统计学意义(P>0.05)。结论:20μmol/L H_2O_2促进斑马鱼侧线毛细胞的正常生理发育,同时亦可促进斑马鱼侧线毛细胞新霉素损伤后再生,但对顺铂损伤后毛细胞再生则无促进作用。
Objective: To investigate the effect of H_2O_2 on the development of hair cells and the regeneration after injury in the lateral line of zebrafish. Methods: The zebrafish 5 days after fertilization were randomly divided into control group, water culture system, H 2 O 2 group, 10 μmol / L and 20 μmol / L H 2 O 2 solution, neomycin group, 200 μmol / L neomycin sulfate The cells were cultured with system water. Neomycin + H 2 O 2 group was treated with 200 μmol / L neomycin sulfate for 1 h and then cultured with 20 μmol / L H 2 O 2 solution. After cisplatin treatment with 1 000 μmol / L cisplatin for 3 h, Platinum + H 2 O 2 group: treated with 1 000 μmol / L cisplatin for 3 h, cultured with 20 μmol / L H 2 O 2 solution. At 0, 24, and 48 h after H 2 O 2 treatment, the lateral hair cells were labeled by immunofluorescence and the counts of three neuructs, P1, P7 and P8, were selected under the fluorescence microscope. The experiment was repeated three times. Results: The numbers of neuronal hair cells of P1, P7 and P8 in 5-7dpf zebrafish were (9.364 ± 0.901, n = 11), (9.645 ± 0.598, n = 15) and (9.922 ± 0.862, (P> 0.05). After treated with 10μmol / L and 20μmol / L H 2 O 2 for 5 days, the numbers of lateral hair cells were 11.540 ± 0.741 and 11.905 ± 0.607, respectively. Compared with the control group (10.841 ± 0.389 ) (P <0.05). The number of hair cells in neomycin + H_2O_248h group and neomycin 48h group were 10.600 ± 0.689 and 8.767 ± 0.603, respectively, there was significant difference between the two groups (P < 0.01). The number of hair cells in cisplatin + H_2O_248h group and cisplatin 48h group was 5.967 ± 1.086,5.633 ± 1.548, respectively. There was no significant difference between the two groups (P> 0.05). CONCLUSION: 20μmol / L H 2 O 2 can promote the normal physiological development of hair cells on the lateral line of zebrafish, and also promote the regeneration of neomycin in the lateral line of zebrafish, but not on the regeneration of hair cells after cisplatin injury.