促甲狀腺激素釋放激素結合高壓氧對大鼠神經病理性痛治療作用的研究
發(fā)布時間：2018-08-16 18:57
【摘要】：目的:從動物行為學以及神經病理性痛的分子生物標志物出發(fā),研究促甲狀腺激素釋放激素(thyrotropin-releasing hormone)與高壓氧(Hyperbaric Oxygen)聯(lián)合后對大鼠神經病理性痛的治療作用,為最終的臨床應用提供科學依據(jù)。方法:(1)將50只大鼠分為假手術組(Sham組)、Charge Coupled Image模型組(CCI組)、促甲狀腺激素釋放激素(thyrotropin-releasing hormone)治療組(CCI+TRH組)、高壓氧(Hyperbaric Oxygen)治療組(CCI+HBO)以及聯(lián)合治療組(CCI+HBO+TRH),通過對每個實驗組大鼠的50%機械刺激縮足反射閾值和熱刺激縮足潛伏期的測定來檢測行為的方法,觀察神經病理性痛大鼠的熱、機械痛閾值在不同治療方法中的變化。(2)通過免疫組化對本研究各分組中神經病理性痛大鼠脊髓背角淺層(I-II)c-Fos的陽性表達情況以及星形膠質細胞活化狀態(tài)進行測定;運用western blot法檢測在不同時間點中各處理組的神經病理性痛大鼠脊髓GFAP蛋白的表達水平。結果:(1)TRH、HBO和HBO+TRH治療均能有效緩解CCI大鼠的神經病理性痛,但HBO+TRH治療的效果更為顯著。(2)相較于S組,CCI引起的神經病理性痛大鼠脊髓背角淺層c-Fos的陽性表達明顯增多;TRH、HBO和HBO+TRH治療均能有效減少大鼠脊髓背角淺層c-Fos陽性細胞核數(shù)量,但HBO+TRH組c-Fos陽性細胞核數(shù)量下降最多;大鼠脊髓背角淺層星形膠質細胞能夠被坐骨神經壓榨激活,相較于CCI組,TRH、HBO和HBO+TRH治療均能有效降低CCI大鼠術后第14、28天的脊髓背角淺層的GFAP陽性細胞核數(shù)量,但HBO+TRH治療組抑制率最高。(3)western blot得到的結果同免疫組化是相同的,相較于CCI組,TRH、HBO和HBO+TRH治療均能有效抑制CCI大鼠脊髓GFAP蛋白的表達,但HBO+TRH治療組抑制率最高。結論:(1)TRH結合HBO治療神經病理性痛最為有效,可長時程降低CCI大鼠熱痛敏和機械性痛覺超敏的嚴重程度,縮短病程。(2)TRH結合HBO治療能更有效的抑制CCI引起的c-Fos表達量增加,同時還能能夠有效抑制術側脊髓背角星形膠質細胞活化,從分子生物標志物水平進一步驗證了TRH結合HBO治療的鎮(zhèn)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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R614
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