甲胺磷对母鸡坐骨神经中神经丝蛋白的影响

Alterations of neurofilament proteins in sciatic nerve of hens induced the delayed neurotoxicity by methamidophos

目的观察甲胺磷诱导母鸡迟发性神经毒性时坐骨神经中神经丝蛋白的变化。方法皮下注射甲胺磷30mg/kg,连续15d。分别在母鸡呈现迟发性神经毒性症状的第2天、第10天和第23天,采用免疫印记法检测坐骨神经沉淀和上清液中神经丝亚单位高分子量神经丝(NF H)、中分子量神经丝(NF M)和低分子量神经丝(NF L)的含量。结果坐骨神经沉淀中,NF H积分密度(IOD)值在呈现症状第2天为145117±17038,比对照组的78875±22569升高了84%,呈现症状第10天和第23天分别为55917±17333和45038±6662,与对照组比较,分别降低了29%和43%;NF M的IOD值在呈现症状第2天时为31493±4625,与对照组的27925±2660比较,差异无统计学意义;呈现症状第10天和第23天分别为19367±2746和6612±1119,明显低于对照组;NF L的IOD值在呈现症状第2天时为39211±3800,明显高于对照组的28749±9319;第10天和第23天分别为27974±3611和21507±2286,与对照组比较差异无统计学意义。3个时间点上清液中NF H的IOD值分别为4709±1739、12337±3205和16745±931,均明显低于对照组的44083±6895;NF M含量明显降低,呈现症状第2天的IOD值为3196±269,比对照组的17243±3232降低了81%,第10天时仅检测到微量,第23天时的IOD值为5206±1292,比对照组降低了70%;

Objective To investigate the dynamic alterations of neurofilament subunits (NF) in sciatic nerve of hens with organophosphorus ester induced the delayed neurotoxicity or neuropathy (OPIDN).Methods Hens with OPIDN were produced by giving 30 mg/kg methamidophos subcutaneously to the 10-month-old Roman hens daily for 15 days, and sacrificed after manifesting neurotoxic clinical signs on the 2nd, 10th, and 23rd day respectively. The sciatic nerves were dissected, homogenized and centrifuged. The levels of NF in supernatant and pellet of sciatic nerves were examined by Western blotting respectively at different time from 2 to 23 days.Results Integrated optional density (IOD) of high molecular weight neurofilament (NF-H) in sciatic nerve pellet of hens on the day 2,10, 23 after appearance of OPIDN were 145 117±17 038, 55 917±17 333 and 45 038±6 662 respectively. As compared with the control group (78 875±22 569), the contents of NF-H in pellet were increased by 84% on day 2, and decreased by 29% and 43% on day 10 and 23 respectively. IOD of NF-H in supernatant of sciatic nerves were 4 709±1 739, 12 337±3 205 and 16 745±931, which were reduced significantly as compared with the control (44 083±6 895)at three different times. There was no significant difference in IOD of middle molecular weight neurofilament (NF-M) between control group (27 925±2 660) and on day 2 (31 493±4 625) in pellet. Those were 19 367±2 746 and 6 612±1 119 respectively on day 10 and day 23 in pellet of hen’s sciatic nerve, which were much less than that in control. Little were detected in supernatant on day 10, and the IOD of NF-M were 3 196±269 and 5 206±1 292 on day 2 and day 23 respectively, which were lessened by 81% and 70% as compared with the control (17 243±3 232).In sciatic nerve pellet of hens, IOD of low molecular weight neurofilament (NF-L) on day 2 was 39 211±3 800, which was much higher than that in the control (28 749±9 319). There were no significant differences between IOD on day 10 ( 27 974± 3 611), day 23 (21 507±2 286) and the control. There was no detection both on day 2 and 10 in supernatant of sciatic nerve, and IOD of NF-L were 5 962±1 929 on day 23,which were reduced significantly compared with the control (11 897±352).Conclusion The alterations of NF in sciatic nerve might contribute to the occurrence and development of OPIDN.