咪康唑对OPIDN的影响及作用机制研究

发布时间：2018-06-01 02:27

本文选题：咪康唑 + OPIDN　；参考：《河北大学》2017年硕士论文

【摘要】：一些有机磷化合物(organophosphorus compound,OP)能够引发一种迟发性的神经病症,称为有机磷引起的迟发性神经病(organophosphate-induced delayed neuropathy,OPIDN)。虽然OPIDN事件报道以来已经过了一个多世纪,但是其作用机理仍然不清楚,更没有有效的治疗方法。本研究旨在研究咪康唑对OPIDN的影响及作用机理。我们以研究OPIDN常用的成年鸡为研究对象,用有机磷化合物磷酸三邻甲苯酯(tri-ocresyl phosphate,TOCP)为诱导剂,从症状得分、组织病理变化和维持髓鞘相关的信号蛋白等多方面探索咪康唑对OPIDN的影响及相关机理。首先将成年鸡分为三组(对照组;TOCP组;TOCP+咪康唑组),对照组口服空胶囊,TOCP组和TOCP+咪康唑组给予TOCP(750 mg/kg,po)处理,TOCP+咪康唑组从TOCP染毒第7天开始直到第21天连续腹腔注射咪康唑(3.5 mg/kg/days)。观察TOCP单独处理组发现从第8天开始成年鸡出现轻微的共济失调,随着时间的推移毒性症状加重(从中度而明显的不协调到严重性、经常性的站立困难),到最后完全瘫痪,根据症状观察可以得出OPIDN模型诱导成功。与TOCP单独处理相比,TOCP处理后再给予咪康唑能够明显改善TOCP引起的体重降低和毒性症状;对脊髓运动神经元尼氏体检测发现,与对照组相比,TOCP单独处理组尼氏体降低约40%,与TOCP单独处理组相比,TOCP+咪康唑组恢复到接近对照水平;对坐骨神经髓鞘特异性蛋白S100β进行免疫荧光染色,与对照组相比,TOCP单独处理组引起坐骨神经S100β明显减少,与TOCP单独处理组相比,TOCP+咪康唑组恢复到接近对照水平;超薄切片观察脊髓和坐骨神经髓鞘的完整性发现,与对照组相比较,TOCP单独处理组坐骨神经髓鞘损伤明显,而咪康唑能够明显改善TOCP引起的这种损伤。为了进一步探索咪康唑如何改善OPIDN,已知ErbB3/Akt和JNK/c-Jun和p38对于维持髓鞘的稳定至关重要。因此,我们运用蛋白免疫印迹的方法检测坐骨神经和脊髓组织ErbB3/Akt、JNK/c-Jun和p38以及维持髓鞘稳定的结构蛋白P0和MBP的变化。结果表明,与对照组相比,单独给予TOCP处理引起脊髓和坐骨神经ErbB3/Akt信号通路激活,脊髓组织JNK激活而坐骨神经JNK抑制,并引起髓磷脂碱性蛋白(myelin basic protein,MBP)的明显下降。咪康唑可以抑制TOCP引起的ErbB3/Akt信号通路激活,能够恢复TOCP引起的脊髓组织JNK激活和坐骨神经JNK抑制,并且能恢复MBP表达水平。我们以sNF96.2雪旺氏细胞为模型进一步探讨咪康唑缓解TOCP引起的迟发性神经毒性作用机理,实验分为四组:DMSO溶剂对照组;咪康唑单独处理组;TOCP单独处理组;TOCP+咪康唑组。用免疫印迹的方法检测各个处理组ErbB3/Akt信号通路以及MBP的变化,结果显示,与对照组相比,单独给予TOCP处理引起雪旺氏细胞ErbB3/Akt信号通路激活以及MBP表达量明显下降,TOCP与咪康唑共同处理可以抑制TOCP引起的ErbB3/Akt信号通路激活,且能够恢复MBP表达水平。综上所述,可以得出咪康唑显著改善了由TOCP引起的成年鸡脊髓和坐骨神经的毒性症状和组织病理损伤。咪康唑可能通过调控ErbB3/Akt信号通路和JNK表达量来改善OPIDN。
[Abstract]:Some organophosphorus compounds (organophosphorus compound, OP) can cause a delayed neuropathy, known as the organophosphate-induced delayed neuropathy (OPIDN) caused by organophosphorus (OPIDN). Although the OPIDN event has been reported for more than a century, its mechanism is still unclear and less effective. The purpose of this study was to study the effect and mechanism of miconazole on OPIDN. We studied the adult chicken in OPIDN, using organophosphorus phosphate three o toluene (tri-ocresyl phosphate, TOCP) as an inducer, from the symptom score, the tissue disease change and the maintenance of the myelin related signal protein. To explore the effect of miconazole on OPIDN and the related mechanism. First, the adult chickens were divided into three groups (control group, TOCP group, TOCP+ miconazole group), the control group was taken orally empty capsules, the TOCP group and TOCP+ miconazole group were treated with TOCP (750 mg/kg, PO), and the miconazole group began to intraperitoneally injected with midazolazole (3.5 mg/kg/days) from TOCP for seventh days from TOCP. A single treatment group of TOCP found a slight ataxia in the adult chicken from eighth days, and with the passage of time the toxic symptoms increased (from moderate to severe incongruity to severity, frequent standing difficulties), to the final complete paralysis, and the OPIDN model could be induced to be successful according to the symptom observation. Compared with TOCP alone, TOC When treated with P, imidazole could significantly improve the weight loss and toxic symptoms caused by TOCP. The Nissl body detection of spinal motor neurons found that, compared with the control group, the Nissl body decreased by about 40% in the TOCP alone group. Compared with the TOCP alone group, the TOCP+ miconazole group recovered to the control level, and the myelin sheath specificity of the sciatic nerve was specific. Protein S100 beta was stained with immunofluorescence. Compared with the control group, the S100 beta of the sciatic nerve decreased significantly in the TOCP alone group. Compared with the TOCP alone group, the TOCP+ miconazole group recovered to the control level, and the ultrathin section observed the integrity of the myelin myelin sheath and the sciatic nerve. Compared with the control group, TOCP alone treated the sciatic group. In order to further explore how miconazole improves the OPIDN, the known ErbB3/Akt and JNK/c-Jun and p38 are essential for maintaining the stability of the myelin sheath. Therefore, we use the method of protein immunoblotting to detect the ErbB3/Akt in the sciatic and spinal cord tissue, JNK/c-Ju, JNK/c-Ju, JNK/c-Ju, JNK/c-Ju, and p38. The changes in N and p38 and the maintenance of myelin stable structural protein P0 and MBP showed that, compared with the control group, the ErbB3/Akt signaling pathway of the spinal cord and sciatic nerve was activated by TOCP treatment alone, the JNK activation of the spinal cord and the inhibition of the JNK of the sciatic nerve, and the obvious decrease of the myelin alkaline egg white (myelin basic protein, MBP). Azole can inhibit the activation of the ErbB3/Akt signaling pathway induced by TOCP, restore the JNK activation of the spinal cord and the JNK inhibition of the sciatic nerve caused by TOCP, and restore the MBP expression level. We use sNF96.2 Schwann's cell as a model to further explore the mechanism of delayed neurotoxicity induced by miconazole in TOCP, and the experiment is divided into four groups: DMSO The solvent control group, miconazole alone treatment group, TOCP alone treatment group and TOCP+ miconazole group. The changes of ErbB3/Akt signaling pathway and MBP in each treatment group were detected by immunoblotting. The results showed that the ErbB3/Akt signaling pathway activation in Schwann cells and the MBP expression decreased significantly by TOCP treatment compared with the control group, TO. CP combined with miconazole can inhibit the activation of the ErbB3/Akt signaling pathway induced by TOCP and can restore the level of MBP expression. In summary, miconazole can significantly improve the toxic symptoms and histopathological damage of the spinal cord and sciatic nerve of adult chickens caused by TOCP. Miconazole may regulate the ErbB3/Akt signaling pathway and the JNK table. Amount to improve OPIDN.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R747.9

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