本文關(guān)鍵詞： 氯化血紅素 腦紅蛋白 脊髓損傷 大鼠　出處：《中國(guó)人民解放軍醫(yī)學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：制作大鼠Allen's脊髓損傷(Spinal Cord Injury, SCI)模型,腹腔注射氯化血紅素(Hemin)溶液。分析Hemin干預(yù)脊髓損傷后大鼠行為學(xué)評(píng)分的變化和不同分組實(shí)驗(yàn)后一定時(shí)間點(diǎn)評(píng)分的差異,Hemin處理后大鼠脊髓內(nèi)腦紅蛋白(Neuroglobin, Ngb)表達(dá)空間和數(shù)量的規(guī)律,以及Hemin處理引起的脊髓Ngb高表達(dá)對(duì)于脊髓損傷后脊髓組織結(jié)構(gòu)、細(xì)胞水平、病理進(jìn)程影響,試為臨床采用Hemin干預(yù)脊髓損傷提供可靠的實(shí)驗(yàn)室理論基礎(chǔ)。方法：健康雄性SD大鼠90只,體重180g-200g,由軍事醫(yī)學(xué)科學(xué)院提供,隨機(jī)分為三組：對(duì)照組(假手術(shù)組)、手術(shù)組(僅造模)、Hemin處理組(造模后腹腔注射Hemin溶液)。假手術(shù)組僅進(jìn)行椎板卸下后還納,不進(jìn)行脊髓打擊,造模后腹腔注射相同量生理鹽水;手術(shù)組采用Allen's法制作大鼠損傷模型,造模后腹腔注射相同量生理鹽水;Hemin處理組進(jìn)行造模和相同量的Hemin溶液腹腔注射。采用大鼠后肢功能BBB評(píng)分、蘇木素伊紅染色、尼氏染色、免疫組化等方法評(píng)價(jià)各組大鼠實(shí)驗(yàn)后1d、3d、7d、14d、28d、56d時(shí)大鼠運(yùn)動(dòng)功能差異、脊髓結(jié)構(gòu)改變、Ngb表達(dá)含量和分布的差異等。實(shí)驗(yàn)相關(guān)結(jié)果數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析。結(jié)果：(1)BBB評(píng)分結(jié)果：對(duì)照組Hemin處理組手術(shù)組。(2)HE染色結(jié)果：對(duì)照組脊髓組織完整,無(wú)明顯的紅細(xì)胞積聚及空隙,無(wú)炎性細(xì)胞浸潤(rùn),神經(jīng)細(xì)胞未出現(xiàn)病理改變;手術(shù)組脊髓具有明顯的紅細(xì)胞積聚及空隙,炎性細(xì)胞浸潤(rùn)明顯,噬神經(jīng)現(xiàn)象等病理改變嚴(yán)重;Hemin處理組損傷重于對(duì)照組,但明顯輕于手術(shù)組,有少量出血和空腔,神經(jīng)組織炎性細(xì)胞浸潤(rùn)現(xiàn)象較少,神經(jīng)細(xì)胞水腫變性較少。(3)尼氏染色結(jié)果：對(duì)照組神經(jīng)元形態(tài)正常,尼氏體圍繞胞核呈顆粒狀,顏色較深切且分布較多,由內(nèi)向外顏色逐漸遞減;Hemin處理組染色結(jié)果中水腫及顏色變淺、數(shù)量減少重于對(duì)照組但輕于手術(shù)組;手術(shù)組神經(jīng)元病理變化最重,除上述情況外,更有尼氏體溶解的現(xiàn)象。(4)免疫組化結(jié)果：Hemin處理組陽(yáng)性細(xì)胞最多,手術(shù)組次之,假手術(shù)組最少。結(jié)論：本實(shí)驗(yàn)通過(guò)采用Hemin腹腔注射干預(yù)脊髓損傷Allen's造模后的SD大鼠,對(duì)脊髓損傷引起脊髓組織Ngb高表達(dá)及Hemin誘導(dǎo)脊髓組織Ngb高表達(dá)的作用進(jìn)行了驗(yàn)證。對(duì)Hemin誘導(dǎo)的脊髓組織Ngb高表達(dá)在脊髓損傷過(guò)程中的作用進(jìn)行了研究和探討。本實(shí)驗(yàn)的結(jié)果提示利用Hemin誘導(dǎo)脊髓Ngb高表達(dá)可以在提高大鼠對(duì)脊髓損傷的抗性。Ngb的神經(jīng)保護(hù)作用涉及多途徑、多環(huán)節(jié),利用Hemin可以增強(qiáng)內(nèi)源性Ngb的表達(dá),從而提高其神經(jīng)保護(hù)作用,這將為SCI的臨床干預(yù)新的思路和藥物選擇提供可靠的實(shí)驗(yàn)室基礎(chǔ)。
[Abstract]:Objective: to establish Allen's rat spinal cord injury (Spinal Cord, Injury, SCI) model, intraperitoneal injection of hemin (Hemin) solution. The difference analysis of the changes of Hemin intervention in rats after spinal cord injury behavior score and different experimental groups after a certain time point score, after the treatment of Hemin in spinal cord of rat Neuroglobin (Neuroglobin Ngb) expression space and number of rules, and Hemin induced high expression of Ngb in spinal cord tissue structure, cell level after spinal cord injury, pathological process, Hemin intervention of spinal cord injury provided theoretical basis for the reliable laboratory for clinical use. Methods: 90 healthy male SD rats, weight 180g-200g, from the Military Medical Science Academy of the PLA provide, randomly divided into three groups: control group (sham operation group), operation group (only model), Hemin group (model after intraperitoneal injection of Hemin solution). The sham operation group underwent laminectomy after discharge Accept, without spinal cord injury, rats after intraperitoneal injection of the same amount of normal saline; operation group by the method of Allen's injury model of rat model after intraperitoneal injection of the same amount of saline; Hemin treatment groups were modeled and the same amount of Hemin solution by intraperitoneal injection. Rat limb function after hematoxylin BBB score. With red staining, Nissl staining, immunohistochemistry methods were evaluated in rats after 1D, 3D, 7d, 14d, 28d, 56d between the motor function of rats, spinal cord structure change, the difference of Ngb expression content and distribution. The experimental results related data were statistically analyzed. Results: (1) results of BBB: control group Hemin group operation group. (2) the results of HE staining: control group of spinal cord tissue integrity, no accumulation of red cells and the gap is obvious, no infiltration of inflammatory cells, nerve cells did not appear pathological changes; spinal surgery group has obvious and accumulation of red blood cells Void, inflammatory cell infiltration was pathological changes serious neuronophagia phenomenon; Hemin treatment group injury in control group, but was lower in the operation group, a small amount of bleeding and cavity, less nerve tissue infiltration of inflammatory cells, nerve cell edema less. (3) Nissl staining results: the control group of neurons normal morphology, Nissl bodies around the nucleus is granular and color is deep and the distribution is more, from the inside to the outside color gradually decreased; Hemin staining results in the treatment group the edema and pale, heavier than the control group but lighter than the operation group to reduce the number of operation group; pathological changes of the neurons, in addition to the above, more chromatolysis phenomenon. (4) the results of immunohistochemistry: Hemin positive cells in treatment group, operation group and sham operation group at least. Conclusion: this experiment by using SD rats by intraperitoneal injection of Hemin intervention for spinal cord injury after Allen's, To cause the high expression of Ngb in spinal cord induced high expression of Ngb and Hemin in spinal cord injury was verified. On Hemin induced spinal cord tissue high expression of Ngb in spinal cord injury in rats was investigated and discussed. The experimental results suggest that the high expression of Ngb in rat spinal cord can improve the neuroprotective effect on rats the resistance of.Ngb spinal cord injury involving multi way, multi link using Hemin, using Hemin can enhance the expression of endogenous Ngb, so as to improve its neuroprotective effect, which will provide SCI clinical intervention ideas and new drugs provide reliable laboratory basis.

【學(xué)位授予單位】：中國(guó)人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2

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本文編號(hào)：1464280