本文關鍵詞： 氯化血紅素 腦紅蛋白 脊髓損傷 大鼠　出處：《中國人民解放軍醫(yī)學院》2015年碩士論文　論文類型：學位論文

【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干預脊髓損傷后大鼠行為學評分的變化和不同分組實驗后一定時間點評分的差異,Hemin處理后大鼠脊髓內(nèi)腦紅蛋白(Neuroglobin, Ngb)表達空間和數(shù)量的規(guī)律,以及Hemin處理引起的脊髓Ngb高表達對于脊髓損傷后脊髓組織結構、細胞水平、病理進程影響,試為臨床采用Hemin干預脊髓損傷提供可靠的實驗室理論基礎。方法：健康雄性SD大鼠90只,體重180g-200g,由軍事醫(yī)學科學院提供,隨機分為三組：對照組(假手術組)、手術組(僅造模)、Hemin處理組(造模后腹腔注射Hemin溶液)。假手術組僅進行椎板卸下后還納,不進行脊髓打擊,造模后腹腔注射相同量生理鹽水;手術組采用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評分、蘇木素伊紅染色、尼氏染色、免疫組化等方法評價各組大鼠實驗后1d、3d、7d、14d、28d、56d時大鼠運動功能差異、脊髓結構改變、Ngb表達含量和分布的差異等。實驗相關結果數(shù)據(jù)進行統(tǒng)計學分析。結果：(1)BBB評分結果：對照組Hemin處理組手術組。(2)HE染色結果：對照組脊髓組織完整,無明顯的紅細胞積聚及空隙,無炎性細胞浸潤,神經(jīng)細胞未出現(xiàn)病理改變;手術組脊髓具有明顯的紅細胞積聚及空隙,炎性細胞浸潤明顯,噬神經(jīng)現(xiàn)象等病理改變嚴重;Hemin處理組損傷重于對照組,但明顯輕于手術組,有少量出血和空腔,神經(jīng)組織炎性細胞浸潤現(xiàn)象較少,神經(jīng)細胞水腫變性較少。(3)尼氏染色結果：對照組神經(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處理組染色結果中水腫及顏色變淺、數(shù)量減少重于對照組但輕于手術組;手術組神經(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)免疫組化結果：Hemin處理組陽性細胞最多,手術組次之,假手術組最少。結論：本實驗通過采用Hemin腹腔注射干預脊髓損傷Allen's造模后的SD大鼠,對脊髓損傷引起脊髓組織Ngb高表達及Hemin誘導脊髓組織Ngb高表達的作用進行了驗證。對Hemin誘導的脊髓組織Ngb高表達在脊髓損傷過程中的作用進行了研究和探討。本實驗的結果提示利用Hemin誘導脊髓Ngb高表達可以在提高大鼠對脊髓損傷的抗性。Ngb的神經(jīng)保護作用涉及多途徑、多環(huán)節(jié),利用Hemin可以增強內(nèi)源性Ngb的表達,從而提高其神經(jīng)保護作用,這將為SCI的臨床干預新的思路和藥物選擇提供可靠的實驗室基礎。
[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.

【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.2

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本文編號：1464280