本文選題：脊髓損傷 + 去鐵胺��； 參考：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：【目的】脊髓損傷是骨科嚴(yán)重的創(chuàng)傷性疾病,目前無有效的治療方式,給患者、家庭和社會帶來嚴(yán)重負(fù)擔(dān)。脊髓損傷后的炎癥反應(yīng)在繼發(fā)性脊髓損傷過程中起到至關(guān)重要的作用,有研究報道去鐵胺(Deferoxamine,DFO)對大鼠脊髓損傷具有一定的治療作用,但是機(jī)制未完全闡明,本研究旨在探索DFO是否在大鼠脊髓打擊損傷模型中對脊髓損傷后炎癥反應(yīng)起抑制作用而對脊髓損傷(Spinal Cord Injury,SCI)產(chǎn)生治療作用�！痉椒ā�1.動物模型制作本研究采用Impactor Model-Ⅱ型脊髓損傷打擊器(10 g×25 mm)制作大鼠脊髓胸10(T10)節(jié)段脊髓挫傷模型。2.動物實驗分組實驗動物采用8周齡雌性Wistar大鼠54只,實驗動物隨機(jī)均分為三組:Sham組,Injury組,Injury+DFO組。3.DFO治療作用評價3.1.總鐵離子含量檢測于SCI后2 d和14 d 2個時間點對3個實驗組脊髓使用總鐵離子檢測試劑盒檢測局部總鐵離子含量。3.2.炎癥因子表達(dá)檢測于SCI后2 d和14 d 2個時間點對3個實驗組脊髓進(jìn)行取材,Western Blot檢測炎癥因子腫瘤壞死因子(TNF-α)、白介素1-β(IL1-β)和凋亡蛋白Caspase-3的表達(dá)情況。3.3.脊髓病理結(jié)構(gòu)評價于SCI后2 d和14 d 2個時間點對各實驗組脊髓進(jìn)行取材并制備脊髓石蠟切片,進(jìn)行膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein,GFAP)、神經(jīng)元核心抗原(neuronal nuclei,NeuN)、2'3'環(huán)核苷酸磷酸二酯酶3(2'3'cyclic nucleotide3'phosphodiesterase,CNPase)免疫組織化學(xué)染色、蘇木精-伊紅染色(HE染色)和TUNEL法凋亡檢測判斷脊髓局部星形膠質(zhì)細(xì)胞、神經(jīng)元及少突膠質(zhì)細(xì)胞、局部組織結(jié)構(gòu)變化和局部細(xì)胞凋亡變化情況。3.4.大鼠后肢運(yùn)動功能評價造模成功后,各實驗組大鼠于給藥后1 d、7 d、14 d、28 d、42 d、56 d使用BBB Scale對大鼠后肢運(yùn)動功能恢復(fù)評分�！窘Y(jié)果】1.成功建立大鼠脊髓挫傷模型造模后大鼠出現(xiàn)擺尾、后肢抽搐、脊髓打擊部位出現(xiàn)紅腫及出血。BBB評分顯示在損傷后1 d所有實驗組大鼠評分均為0分。2.總鐵離子含量檢測SCI后2 d,Sham組、Injury組和Injury+DFO組的總鐵離子含量分別為8.930±2.124(μmol/gprot)、22.502±3.436(μmol/gprot)和9.141±1.062(μmol/gprot)。SCI 14 d后Sham組、Injury組和Injury+DFO組的總鐵離子含量分別為8.785±2.671(μmol/gprot)、65.677±2.604(μmol/gprot)和13.009±1.272(μmol/gprot)。3.炎癥因子表達(dá)檢測SCI后2d,Sham組、Injury組和Injury+DFO組SCI局部IL1-β的相對表達(dá)水平分別為1±0.231、1.74±0.217和1.01±0.312;Sham組、Injury組和Injury+DFO組SCI局部TNF-α的相對表達(dá)水平分別為1±0.193、1.53±0.245和1.18±0.187;Sham組、Injury組和Injury+DFO組SCI局部Caspase-3相對表達(dá)水平分別為1±0.159、2.89±0.231和1.71±0.224。SCI 14d后,Sham組、Injury組和Injury+DFO組SCI局部IL1-β的相對表達(dá)水平分別為1±0.267、1.73±0.193和0.93±0.287;Sham組、Injury組和Injury+DFO組SCI局部TNFα的相對表達(dá)水平分別為1.02±0.291、1.39±0.177和0.97±0.169;Sham組、Injury組和Injury+DFO組SCI局部Caspase-3的相對表達(dá)水平分別為1±0.179、2.27±0.312和1.20±0.254。4.脊髓病理評價于SCI后2 d和14 d 2個時間點對各實驗組脊髓進(jìn)行取材并制備脊髓切片,進(jìn)行膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein,GFAP)、神經(jīng)元核心抗原(Neuronal nuclei,Neu N)、2'3'環(huán)核苷酸磷酸二酯酶3(2'3'cyclic nucleotide 3'phosphodiesterase,CNPase)免疫組織化學(xué)染色、蘇木精-伊紅染色(HE染色)和TUNEL法凋亡染色,結(jié)果顯示:相對于Injury組,Injury+DFO組明顯的減少GFAP面積和TUNEL陽性細(xì)胞數(shù)量,并且增加CNPase陽性面積和NeuN陽性細(xì)胞數(shù)量,對SCI后脊髓病理結(jié)構(gòu)有明顯的修復(fù)作用。5.大鼠后肢運(yùn)動功能評價Sham組在第0天的BBB評分為21分,而Injury和Injury+DFO組的評分均為0分,表明大鼠SCI模型的有效性。各組評分隨時間逐漸升高,然而,在SCI后7天,Injury和Injury+DFO組之間的BBB評分具有統(tǒng)計學(xué)顯著差異。在SCI后8周,Injury組的BBB評分為14,Injury+DFO組為18。結(jié)果表明,DFO對大鼠SCI后運(yùn)動功能有明顯的恢復(fù)作用�！窘Y(jié)論】DFO可以顯著減少SCI局部鐵離子含量,減少SCI局部TNFα、IL1-β和Caspase-3等因子的表達(dá),保護(hù)局部原有結(jié)構(gòu)細(xì)胞,減少細(xì)胞凋亡,恢復(fù)大鼠后肢運(yùn)動功能,促進(jìn)SCI的修復(fù),為SCI治療提供新的藥物治療選擇,為臨床修復(fù)SCI提供實驗依據(jù)。
[Abstract]:[Objective] spinal cord injury is a serious traumatic disease in the Department of orthopedics. There is no effective treatment for patients, family and society. The inflammatory reaction after spinal cord injury plays a vital role in secondary spinal cord injury. It is reported that Deferoxamine (DFO) has a certain effect on spinal cord injury in rats. The therapeutic effect, but the mechanism is not fully elucidated, the purpose of this study is to explore the effect of DFO on spinal cord injury (Spinal Cord Injury, SCI) in the spinal cord injury injury model in rats. [Methods] 1. animal models were made to use Impactor Model- II type spinal cord injury. The rat spinal cord thoracic 10 (T10) (T10) segment spinal contusion model was made by the attack device (10 G x 25 mm) in the animal experiment group of.2. animal experiment group and 54 female Wistar rats of 8 weeks of age were used. The experimental animals were randomly divided into three groups: Sham group, Injury group and Injury+DFO group.3.DFO therapeutic effect evaluation 3.1. total iron content of 3.1. was detected in 3 after SCI 2 D and 14 times 2 time points. In the experimental group, the total iron ion content of the spinal cord was detected by the total iron ion detection kit and the expression of.3.2. inflammatory factor was detected in the spinal cord of 3 experimental groups after SCI 2 D and 14 d. Western Blot was used to detect the inflammatory factor tumor necrosis factor (TNF- alpha), and the expression of interleukin 1- beta (IL1- beta) and apoptotic protein Caspase-3 was.3.3.. The spinal cord pathological structure was evaluated at the 2 time points of SCI 2 D and 14 d to prepare the spinal cord and prepare the spinal paraffin section for glial fibrillary acidic protein (glial fibrillary acidic protein, GFAP), neuron core antigen (neuronal nuclei, NeuN), and 2'3'cyclic nucleotide phosphodiesterase 3. Erase, CNPase) immuno histochemical staining, hematoxylin eosin staining (HE staining) and TUNEL apoptosis detection to determine the local astrocytes, neurons and oligodendrocytes in the spinal cord, the changes of local tissue structure and the changes of local cell apoptosis in the.3.4. rat hind limb movement function evaluation, the rats in the experimental group were given the drug after administration. 1 D, 7 d, 14 d, 28 d, 42 d, 56 D used BBB Scale to evaluate the motor function recovery of the hind limbs. [results] 1. successfully established the rat spinal cord contusion model, the rats appeared the tail, the hind limbs twitching, the swelling of the spinal cord and the bleeding.BBB score showed that the scores of the rats in all the experimental groups of the 1 D were 0 fractions and.2. total iron ions after the injury. The total iron content in the 2 D, Sham, Injury and Injury+DFO groups were 8.930 + 2.124 (mu mol/gprot), 22.502 + 3.436 (mu mol/gprot) and 9.141 + 1.062 (mu mol/gprot).SCI 14 d, respectively. The total iron content of the Injury group and the group was 8.785 + 2.671 (MU), 65.677 + 2.604 (MU) and 13.009, respectively. The relative expression level of SCI local IL1- beta in 2D, Sham, Injury and Injury+DFO groups was 1 + 0.231,1.74 + 0.217 and 1.01 + 0.312, respectively, and the relative expression level of the Sham group and Injury+DFO group was 1 + 1 + 0.245 and 1.18 + 0.187 in the Sham group and the Injury+DFO group, respectively. The relative expression level of local Caspase-3 in group SCI and Injury+DFO group was 1 + 0.159,2.89 + 0.231 and 1.71 + 0.224.SCI 14d respectively. The relative expression level of SCI partial IL1- beta in Sham group, Injury group and Injury+DFO group was 1 + 0.193 and 0.93 + 0.287, respectively. The relative expression level of SCI local Caspase-3 in group Sham, Injury and Injury+DFO was 1 + 0.179,2.27 + 0.312 and 1.20 + 0.254.4. in the spinal cord pathology evaluation at 2 time points of 2 D and 14 d after SCI, and the spinal cord sections were prepared and prepared for the spinal cord, and the glial fibrillary acidic protein (glial) was carried out. Brillary acidic protein, GFAP), neuron core antigen (Neuronal nuclei, Neu N), 2'3'cyclic nucleotide phosphodiesterase 3 (2'3'cyclic nucleotide 3'phosphodiesterase), immunohistochemical staining, hematoxylin eosin staining and dying dying staining. The number of FAP area and TUNEL positive cells increased the positive area of CNPase and the number of NeuN positive cells. After SCI, the pathological structure of the spinal cord was obviously repaired in the.5. rat hind limb motor function. The BBB score of the Sham group in the Sham group was 21 at zeroth days, while the scores of the Injury and Injury+DFO groups were all 0 points, indicating the effectiveness of the SCI model of the rats. The score increased with time, however, at 7 days after SCI, the BBB score between the Injury and Injury+DFO groups was statistically significant. At 8 weeks after SCI, the BBB score of group Injury was 14, and Injury+DFO group 18. showed that DFO had a significant recovery of the motor function after SCI in rats. [Conclusion] DFO can significantly reduce the part of the iron ions containing SCI. Quantity, reduce the expression of SCI local TNF alpha, IL1- beta and Caspase-3, protect local original structure cells, reduce cell apoptosis, restore the motor function of the hind limbs of rats, promote the repair of SCI, provide new drug treatment options for the treatment of SCI, and provide experimental basis for the clinical repair of SCI.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R651.2

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