促甲状腺激素释放激素结合高压氧对大鼠神经病理性痛治疗作用的研究
发布时间：2018-08-16 18:57
【摘要】：目的:从动物行为学以及神经病理性痛的分子生物标志物出发,研究促甲状腺激素释放激素(thyrotropin-releasing hormone)与高压氧(Hyperbaric Oxygen)联合后对大鼠神经病理性痛的治疗作用,为最终的临床应用提供科学依据。方法:(1)将50只大鼠分为假手术组(Sham组)、Charge Coupled Image模型组(CCI组)、促甲状腺激素释放激素(thyrotropin-releasing hormone)治疗组(CCI+TRH组)、高压氧(Hyperbaric Oxygen)治疗组(CCI+HBO)以及联合治疗组(CCI+HBO+TRH),通过对每个实验组大鼠的50%机械刺激缩足反射阈值和热刺激缩足潜伏期的测定来检测行为的方法,观察神经病理性痛大鼠的热、机械痛阈值在不同治疗方法中的变化。(2)通过免疫组化对本研究各分组中神经病理性痛大鼠脊髓背角浅层(I-II)c-Fos的阳性表达情况以及星形胶质细胞活化状态进行测定;运用western blot法检测在不同时间点中各处理组的神经病理性痛大鼠脊髓GFAP蛋白的表达水平。结果:(1)TRH、HBO和HBO+TRH治疗均能有效缓解CCI大鼠的神经病理性痛,但HBO+TRH治疗的效果更为显著。(2)相较于S组,CCI引起的神经病理性痛大鼠脊髓背角浅层c-Fos的阳性表达明显增多;TRH、HBO和HBO+TRH治疗均能有效减少大鼠脊髓背角浅层c-Fos阳性细胞核数量,但HBO+TRH组c-Fos阳性细胞核数量下降最多;大鼠脊髓背角浅层星形胶质细胞能够被坐骨神经压榨激活,相较于CCI组,TRH、HBO和HBO+TRH治疗均能有效降低CCI大鼠术后第14、28天的脊髓背角浅层的GFAP阳性细胞核数量,但HBO+TRH治疗组抑制率最高。(3)western blot得到的结果同免疫组化是相同的,相较于CCI组,TRH、HBO和HBO+TRH治疗均能有效抑制CCI大鼠脊髓GFAP蛋白的表达,但HBO+TRH治疗组抑制率最高。结论:(1)TRH结合HBO治疗神经病理性痛最为有效,可长时程降低CCI大鼠热痛敏和机械性痛觉超敏的严重程度,缩短病程。(2)TRH结合HBO治疗能更有效的抑制CCI引起的c-Fos表达量增加,同时还能能够有效抑制术侧脊髓背角星形胶质细胞活化,从分子生物标志物水平进一步验证了TRH结合HBO治疗的镇痛作用。
[Abstract]:Aim: to study the therapeutic effect of thyrotropin releasing hormone (thyrotropin-releasing hormone) combined with hyperbaric oxygen (Hyperbaric Oxygen) on neuropathic pain in rats based on animal behavior and molecular biomarkers of neuropathic pain. To provide scientific basis for the final clinical application. Methods: (1) 50 rats were divided into sham-operation group (Sham group) and charge Coupled Image model group (CCI group), thyrotropin-releasing hormone (thyrotropin-releasing hormone) treatment group (CCI TRH group, hyperbaric oxygen (Hyperbaric Oxygen) treatment group (CCI HBO) and combined treatment group (CCI HBO TRH), through the treatment of each solid. A method for measuring behavior of 50% mechanically stimulated foot contraction reflex threshold and thermal stimulation foot contraction latency in the experimental group. To observe the fever of neuropathic pain rats, The changes of mechanical pain threshold in different treatment methods. (2) the positive expression of I-II c-Fos and the activation of astrocytes in the spinal dorsal horn of neuropathic pain rats were determined by immunohistochemistry. The expression of GFAP protein in spinal cord of rats with neuropathic pain was detected by western blot method at different time points. Results: (1) both TRH-HBO and HBO TRH could relieve neuropathic pain in CCI rats. But the effect of HBO TRH treatment was more significant. (2) compared with S group, the positive expression of c-Fos in the superficial layer of spinal dorsal horn was significantly increased in rats with neuropathic pain. Both HBO TRH and HBO TRH treatment could effectively reduce the number of c-Fos positive nuclei in the superficial spinal cord of rats. However, the number of c-Fos positive nuclei in HBO TRH group decreased the most, and the superficial astrocytes in the dorsal horn of spinal cord were activated by sciatic nerve squeezing. Compared with CCI group, HBO and HBO TRH treatment could effectively reduce the number of GFAP positive nuclei in the superficial layer of spinal dorsal horn of CCI rats on the 14th day after operation, but the inhibitory rate of HBO TRH treatment group was the highest. (3) the results of) western blot were the same as those of immunohistochemistry. Compared with CCI group, both HBO and HBO TRH could effectively inhibit the expression of GFAP protein in spinal cord of CCI rats, but HBO TRH treatment group had the highest inhibition rate. Conclusion: (1) TRH combined with HBO is the most effective in the treatment of neuropathic pain, which can reduce the severity of hyperthermia and mechanical hyperalgesia in CCI rats, and shorten the course of disease. (2) TRH combined with HBO can effectively inhibit the increase of c-Fos expression induced by CCI. At the same time, it can effectively inhibit the activation of astrocytes in the dorsal horn of spinal cord, which further proves the analgesic effect of TRH combined with HBO in the level of molecular biomarker.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R614
[Abstract]:Aim: to study the therapeutic effect of thyrotropin releasing hormone (thyrotropin-releasing hormone) combined with hyperbaric oxygen (Hyperbaric Oxygen) on neuropathic pain in rats based on animal behavior and molecular biomarkers of neuropathic pain. To provide scientific basis for the final clinical application. Methods: (1) 50 rats were divided into sham-operation group (Sham group) and charge Coupled Image model group (CCI group), thyrotropin-releasing hormone (thyrotropin-releasing hormone) treatment group (CCI TRH group, hyperbaric oxygen (Hyperbaric Oxygen) treatment group (CCI HBO) and combined treatment group (CCI HBO TRH), through the treatment of each solid. A method for measuring behavior of 50% mechanically stimulated foot contraction reflex threshold and thermal stimulation foot contraction latency in the experimental group. To observe the fever of neuropathic pain rats, The changes of mechanical pain threshold in different treatment methods. (2) the positive expression of I-II c-Fos and the activation of astrocytes in the spinal dorsal horn of neuropathic pain rats were determined by immunohistochemistry. The expression of GFAP protein in spinal cord of rats with neuropathic pain was detected by western blot method at different time points. Results: (1) both TRH-HBO and HBO TRH could relieve neuropathic pain in CCI rats. But the effect of HBO TRH treatment was more significant. (2) compared with S group, the positive expression of c-Fos in the superficial layer of spinal dorsal horn was significantly increased in rats with neuropathic pain. Both HBO TRH and HBO TRH treatment could effectively reduce the number of c-Fos positive nuclei in the superficial spinal cord of rats. However, the number of c-Fos positive nuclei in HBO TRH group decreased the most, and the superficial astrocytes in the dorsal horn of spinal cord were activated by sciatic nerve squeezing. Compared with CCI group, HBO and HBO TRH treatment could effectively reduce the number of GFAP positive nuclei in the superficial layer of spinal dorsal horn of CCI rats on the 14th day after operation, but the inhibitory rate of HBO TRH treatment group was the highest. (3) the results of) western blot were the same as those of immunohistochemistry. Compared with CCI group, both HBO and HBO TRH could effectively inhibit the expression of GFAP protein in spinal cord of CCI rats, but HBO TRH treatment group had the highest inhibition rate. Conclusion: (1) TRH combined with HBO is the most effective in the treatment of neuropathic pain, which can reduce the severity of hyperthermia and mechanical hyperalgesia in CCI rats, and shorten the course of disease. (2) TRH combined with HBO can effectively inhibit the increase of c-Fos expression induced by CCI. At the same time, it can effectively inhibit the activation of astrocytes in the dorsal horn of spinal cord, which further proves the analgesic effect of TRH combined with HBO in the level of molecular biomarker.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R614
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