臭氧在溶血磷脂酸脱髓鞘神经病理性疼痛中对TNF-α及髓鞘相关糖蛋白作用的研究

Study On The Effect Of Ozone On TNF-α And Myelin Related Glycoproteins In Neuropathic Pain Induced By Demyelinating Of Lysophosphatidic Acid

摘要:

目的：探讨单次椎间孔注射臭氧在溶血磷脂酸（lysophosphatidic acid，LPA）脱髓鞘神经病理性疼痛模型中对背根神经髓鞘相关糖蛋白(myeline-associated glycoprotein MAG）及肿瘤坏死因子α（tumor necrosis factor α，TNF-α）表达的影响。方法：c56雄性小鼠鞘内注射1nmol LPA进行神经病理性疼痛造模，造模后24 小时，将小鼠随机分为人造脑脊液（artificial cerebralspinal fluid，aCSF）（LPA+aCSF组）及臭氧组（LPA+O3组），同节段椎间孔分别单次注射aCSF或臭氧, 在注射LPA前（t0）、注射臭氧或aCSF前（t1）、注射臭氧或aCSF后24小时（t2）、3天（t3）、7天（t4）、14天（t5）时进行疼痛行为学检测，并用蛋白质免疫印迹法在不同时间点（t0-t4）检测背根神经髓鞘蛋白MAG及TNF-α的表达变化。结果：1.与人造脑脊液组对比，臭氧注射后24小时机械痛敏阈值和热痛敏潜伏期显著上升，维持到第7天；2.与人造脑脊液组对比，臭氧注射后24小时TNF-α表达显著下降，3天时最低，并维持到第7天；3.与人造脑脊液组对比，臭氧组注射后24小时、3天MAG的蛋白定量更加稳定，无进一步降解，仅在7天时稍下降。结论：在溶血磷脂酸脱髓鞘神经病理性疼痛中单次椎间孔注射臭氧通过抑制炎症因子释放以及维持MAG稳定性可以明显缓解神经病理性疼痛。

Abstract:

Objective: To investigate the effect of intravertebral injection of single ozone on the expression of myline-associated glycoprotein MAG and tumor necrosis factor α(TNF-α) in the dorsal root nerve in the rational pain model of demyelinating neuropathy by lysophosphatidic acid (LPA).Methods:C56 male mice were injected with 1nmol LPA for neuropathic pain modeling. 24 hours after modeling, mice were randomly divided into artificial cerebralspinal fluid (aCSF) (LPA+aCSF group) and ozone group (LPA+O3 group).With segmental intervertebral foramen aCSF or ozone injection respectively, before the injection of LPA (t0), ozone injection or aCSF (t1), 24 hours after injection of ozone or aCSF (t2), 3 days (t3), 7 days (t4), 14 days (t5) is used to detect the pain behavior, and the protein immunoblot method at different time points (t0 - t4) test dorsal nerve myelin protein MAG and changes the expression of TNF-α.Results: 1. Compared with the artificial cerebrospinal fluid group, the mechanical pain sensitivity threshold and thermal pain sensitivity incubation period increased significantly 24 hours after ozone injection, and remained unchanged until day 7; 2.2. Compared with the artificial cerebrospinal fluid group, the expression of TNF-α was significantly decreased 24 hours after ozone injection, the lowest at 3 days, and maintained until the 7th day;3. Compared with the artificial cerebrospinal fluid group, the protein ration of MAG 24 hours and 3 days after injection in the ozone group was more stable, with no further degradation and only slightly decreased at 7 days.Conclusion: the injection of single ozone into the intervertebral foramen in the demyelinating neuropathic pain of lysophosphatidic acid can significantly relieve the neuropathic pain by inhibiting the release of inflammatory factors and maintaining the stability of MAG.