医用臭氧缓解福尔马林炎性疼痛的研究

发布时间：2018-11-11 17:15

【摘要】：目的:明确医用臭氧(O_3)对福尔马林炎性疼痛大鼠脊髓背角小胶质细胞活化以及TNF-α、IL-1β、MCP-1表达的影响,探究臭氧缓解炎性疼痛的机制。方法:成年雄性SD大鼠随机分为4组:(1)对照组(C组):大鼠足底注射100μl生理盐水;(2)福尔马林组(F组):足底注射100μl 2%福尔马林建立炎性疼痛模型,鞘内注射生理盐水10μl;(3)米诺四环素组(MI组):建立炎性疼痛模型后,鞘内注射MI 10μl(5ug/ul);(4)臭氧组(O_3组):建立炎性疼痛模型后,患侧足掌红肿处局部注射O_3 50μl(30 ug/ml),每组10只大鼠。分别测量大鼠建模前(0h)和给药后(1 h、3 h、12 h、1d、3 d和5 d)机械缩足阈值(mechanical withdrawal threshold,MWT)和热缩足潜伏期(thermal withdrawal latency,TWL)。利用Western blot技术观察大鼠脊髓背角小胶质细胞(1d和5d)特异性标记蛋白OX42表达情况,并运用ELISA法检测大鼠脊髓背角TNF-α、IL-1β和MCP-1的表达量。结果:(1)行为学:C组大鼠各时间点MWT和TWL值无明显变化(P0.05);F组、MI组和O_3组MWT和TWL较基础值(0h)显著下降(P0.05);MI组和O_3组MWT和TWL较F组显著升高(P0.05,n=10)。(2)Western blot:与C组比较,F组、MI组和O_3组OX42表达显著增加(P0.05);与F组比较,MI组和O_3组OX42表达明显降低(P0.05,n=10)。(3)ELISA:与C组比较,F组、MI组和O_3组脊髓背角MCP-1、IL-1β和TNF-α的表达水平显著升高(P0.05);与F组比较,MI组和O_3组脊髓背角MCP-1、IL-1β和TNF-α的表达水平显著降低(P0.05,n=10)。结论:(1)脊髓背角小胶质细胞、TNF-α、IL-1β和MCP-1参与调节福尔马林炎性疼痛;(2)足底注射医用臭氧可显著缓解福尔马林诱导的炎性疼痛;(3)医用臭氧可能通过抑制脊髓背角小胶质细胞活化,降低TNF-α、IL-1β和MCP-1表达,从而减轻福尔马林炎性疼痛大鼠的痛觉反应。
[Abstract]:Aim: to investigate the effects of medical ozone (O _ 3) on the activation of microglia and the expression of TNF- 伪, IL-1 尾, MCP-1 in spinal dorsal horn of rats with formalin inflammatory pain, and to explore the mechanism of ozone relieving inflammatory pain. Methods: adult male SD rats were randomly divided into four groups: (1) control group (group C): rats were injected 100 渭 l physiological saline into plantar; (2) in formalin group (F group), the inflammatory pain model was established by injection of 100 渭 l 2% formalin into the plantar and 10 渭 l of normal saline intrathecally. (3) minocycline group (MI group): after inflammatory pain model was established, MI 10 渭 l (5ug/ul) was injected intrathecally; (4) Ozone group: after the inflammatory pain model was established, 10 rats in each group were given local injection of O 350 渭 l (30 ug/ml) at the redness and swelling of the palmar of the affected side. Mechanical foot contraction threshold (mechanical withdrawal threshold,MWT) and thermal contraction latency (thermal withdrawal latency,TWL) were measured before (0 h) and after administration (1 h, 3 h, 12 h, 1 d and 5 d), respectively. The expression of TNF- 伪, IL-1 尾 and MCP-1 in rat spinal dorsal horn microglia (1 d and 5 d) was observed by Western blot technique, and the expression of TNF- 伪, IL-1 尾 and MCP-1 in spinal dorsal horn was detected by ELISA method. Results: (1) behavior: the values of MWT and TWL in group C did not change significantly at each time point (P0.05). The values of MWT and TWL in group MI and group O were significantly lower than those in group C (P0.05). The expression of OX42 in MI group and O3 group was significantly higher than that in F group (P 0.05). Compared with C group, OX42 expression in F group, MI group and O3 group was significantly higher than that in C group (P 0.05). Compared with group F, the expression of OX42 in MI group and O _ S _ 3 group was significantly lower than that in C group (P 0.05). (3), and in F group, MI group and O _ 3 group, MCP-1, expression in spinal dorsal horn was significantly lower than that in C group. The expression of IL-1 尾 and TNF- 伪 increased significantly (P0.05). Compared with group F, the expression levels of MCP-1,IL-1 尾 and TNF- 伪 in the spinal dorsal horn of MI group and Ostex group were significantly decreased (P0.05 nnti10). Conclusion: (1) the spinal dorsal horn microglia, TNF- 伪, IL-1 尾 and MCP-1 are involved in the regulation of formalin inflammatory pain, (2) the injection of medical ozone into the plantar can significantly relieve the formalin induced inflammatory pain. (3) Medical ozone may reduce the expression of TNF- 伪, IL-1 尾 and MCP-1 by inhibiting the activation of microglia in the dorsal horn of spinal cord, so as to alleviate the pain response of formalin inflammatory pain rats.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614

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