【摘要】：目的:明確醫(yī)用臭氧(O_3)對福爾馬林炎性疼痛大鼠脊髓背角小膠質細胞活化以及TNF-α、IL-1β、MCP-1表達的影響,探究臭氧緩解炎性疼痛的機制。方法:成年雄性SD大鼠隨機分為4組:(1)對照組(C組):大鼠足底注射100μl生理鹽水;(2)福爾馬林組(F組):足底注射100μl 2%福爾馬林建立炎性疼痛模型,鞘內注射生理鹽水10μl;(3)米諾四環(huán)素組(MI組):建立炎性疼痛模型后,鞘內注射MI 10μl(5ug/ul);(4)臭氧組(O_3組):建立炎性疼痛模型后,患側足掌紅腫處局部注射O_3 50μl(30 ug/ml),每組10只大鼠。分別測量大鼠建模前(0h)和給藥后(1 h、3 h、12 h、1d、3 d和5 d)機械縮足閾值(mechanical withdrawal threshold,MWT)和熱縮足潛伏期(thermal withdrawal latency,TWL)。利用Western blot技術觀察大鼠脊髓背角小膠質細胞(1d和5d)特異性標記蛋白OX42表達情況,并運用ELISA法檢測大鼠脊髓背角TNF-α、IL-1β和MCP-1的表達量。結果:(1)行為學:C組大鼠各時間點MWT和TWL值無明顯變化(P0.05);F組、MI組和O_3組MWT和TWL較基礎值(0h)顯著下降(P0.05);MI組和O_3組MWT和TWL較F組顯著升高(P0.05,n=10)。(2)Western blot:與C組比較,F組、MI組和O_3組OX42表達顯著增加(P0.05);與F組比較,MI組和O_3組OX42表達明顯降低(P0.05,n=10)。(3)ELISA:與C組比較,F組、MI組和O_3組脊髓背角MCP-1、IL-1β和TNF-α的表達水平顯著升高(P0.05);與F組比較,MI組和O_3組脊髓背角MCP-1、IL-1β和TNF-α的表達水平顯著降低(P0.05,n=10)。結論:(1)脊髓背角小膠質細胞、TNF-α、IL-1β和MCP-1參與調節(jié)福爾馬林炎性疼痛;(2)足底注射醫(yī)用臭氧可顯著緩解福爾馬林誘導的炎性疼痛;(3)醫(yī)用臭氧可能通過抑制脊髓背角小膠質細胞活化,降低TNF-α、IL-1β和MCP-1表達,從而減輕福爾馬林炎性疼痛大鼠的痛覺反應。
[Abstract]:Aim: to investigate the effects of medical ozone (O _ 3) on the activation of microglia and the expression of TNF- 偽, 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- 偽, 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- 偽, 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- 偽 increased significantly (P0.05). Compared with group F, the expression levels of MCP-1,IL-1 尾 and TNF- 偽 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- 偽, 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- 偽, 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.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R614

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