本文選題：天麻素 + 脊髓損傷��； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]脊髓損傷(spinal cord injury,SCI)是一種嚴(yán)重的創(chuàng)傷性疾病,目前尚缺乏有效的治療手段。SCI后星形膠質(zhì)細(xì)胞活化增生形成膠質(zhì)瘢痕,維持損傷部位脊髓結(jié)構(gòu)的完整性。但過度增生的膠質(zhì)瘢痕會阻礙軸突生長,同時分泌神經(jīng)抑制因子影響神經(jīng)元存活,阻礙神經(jīng)再生。本研究用天麻素(GAS)對鉗夾型急性大鼠脊髓損傷模型進(jìn)行干預(yù),探討SCI后天麻素干預(yù)對膠質(zhì)瘢痕形成的影響以及對脊髓神經(jīng)功能恢復(fù)的作用。[方法]1、80只清潔級成年雄性SD大鼠,體重220g-240g,隨機(jī)分為5組:①假手術(shù)組(Sham組,n=16),②損傷組(SCI組,n=16),③低劑量天麻素治療組(SCI+GAS(L)組,50mg/Kg,n=16),④中劑量天麻素治療組(SCI+GAS(M)組,100mg/Kg,n=16),⑤高劑量天麻素治療組(SCI+GAS(H)組,150mg/Kg,n=16)。建立鉗夾型大鼠SCI模型。其中Sham組僅切除椎板,SCI組和各治療組采用70g動脈瘤夾對大鼠T10節(jié)段脊髓進(jìn)行30秒鉗夾損傷。術(shù)后各治療組分別給予不同濃度天麻素連續(xù)干預(yù)7天,Sham組和SCI組給予等量生理鹽水。分別于術(shù)后1d、3d、7d、14d、21d、28d 采用 Basso Beattie Bresnahan 運(yùn)動功能評分(BBB 運(yùn)動功能評分)和Rivlin斜板實(shí)驗(yàn)評估脊髓神經(jīng)功能恢復(fù)情況。2、損傷后4w進(jìn)行灌注、取材。觀察損傷節(jié)段脊髓形態(tài)變化,常規(guī)脫水、包埋,進(jìn)行冰凍切片。免疫熒光染色后,對圖像進(jìn)行定量分析,測定星形膠質(zhì)細(xì)胞特殊標(biāo)記物膠質(zhì)纖維酸性蛋白(Glial Fibrillary Acidic Protein,GFAP)表達(dá)以及膠質(zhì)瘢痕面積。3、損傷后4w行HE染色檢測脊髓形態(tài)和脊髓空洞面積;行尼氏(Nissl)染色檢測神經(jīng)元尼氏小體數(shù)量、形態(tài)。4、損傷后2w于各組中各隨機(jī)抽取4只大鼠,立體定位下注射熒光素標(biāo)記BDA神經(jīng)纖維示蹤劑。損傷后4w行冰凍切片后在熒光顯微鏡下觀察神經(jīng)軸突數(shù)量,評估脊髓神經(jīng)傳導(dǎo)通路恢復(fù)情況。[結(jié)果]1、術(shù)后各組BBB評分和Rivlin斜板實(shí)驗(yàn)評分評分均下降,隨著術(shù)后時間增長各組評分逐漸升高。Sham組評分于各時相點(diǎn)均高于其余各組(P0.05),各GAS治療組在術(shù)后3d及其后各時相點(diǎn)評分均明顯均高于SCI組(P0.05),SCI+GAS(M)組評分均高于其余兩個天麻素治療組(P0.05)。2、損傷后4w各GAS治療組和SCI組GFAP表達(dá)較Sham組均顯著增高(P0.05),各GAS治療組GFAP表達(dá)明顯低于SCI組,SCI+GAS(M)組GFAP抑制最為明顯,(P0.05)。各GAS治療組膠質(zhì)瘢痕陽性染色面積明顯減小,SCI+GAS(M)組膠質(zhì)瘢痕面積明顯低于SCI組,(P0.05)。3、損傷后4w,SCI組脊髓組織結(jié)構(gòu)排列混亂,脊髓空洞形成,各GAS治療組脊髓結(jié)構(gòu)更為清晰,空洞面積減小,SCI+GAS(M)組脊髓空洞面積明顯低于SCI 組,(P0.05)。SCI+GAS(M)組 Nissl 小體數(shù)量明顯多于 SCI 組,(P0.05)。4、損傷4周后Sham組脊髓BDA陽性染色神經(jīng)纖維最多,各GAS治療組損傷平面陽性染色的神經(jīng)纖維明顯多于SCI組,其中SCI+GAS(M)組陽性染色纖維多于其余兩個治療組,(P0.05)。[結(jié)論]1、GAS可以下調(diào)SCI后損傷區(qū)域星形膠質(zhì)細(xì)胞GFAP表達(dá)水平,抑制星形膠質(zhì)細(xì)胞活化,減輕膠質(zhì)瘢痕的形成。2、GAS可以保護(hù)SCI后損傷區(qū)域神經(jīng)元,減輕脊髓空洞形成,促進(jìn)軸突再生,促使脊髓神經(jīng)傳導(dǎo)通路修復(fù)。3、GAS可以提高大鼠SCI后行為學(xué)評分,促進(jìn)神經(jīng)功能恢復(fù),其治療大鼠SCI效果的最佳給藥濃度范圍可能在100mg/Kg左右。
[Abstract]:[Objective] spinal cord injury (SCI) is a serious traumatic disease. At present, there is still a lack of effective treatment of astrocytes by activation of astrocytes to form glial scar and maintain the integrity of the spinal structure in the injured part of the spinal cord. However, hyperplastic glial scar will impede the growth of axon and secrete the neuroinhibitory factor. In this study, the effects of gastrodin on the formation of glial scar and the function of spinal nerve function recovery after SCI were investigated with gastrodin (GAS) and the effect of gastrodin intervention on the formation of glial scar after SCI. [methods only 5 groups of male SD rats, 220g-240g, were randomly divided into 5 groups: (1) the sham operation group (group Sham, n=16), the injury group (group SCI, n=16), the low dose gastrodin group (SCI+GAS (L), 50mg/Kg, n=16), (SCI+GAS (M), 100mg/Kg, n=16) in the middle dose group of gastrodin (SCI+GAS (M), 100mg/Kg, n=16). Group and treatment group were treated with 70g aneurysm clips for 30 seconds of clamp injury to the T10 segment of the spinal cord of rats. The treatment groups were given different concentrations of gastrodin for 7 days, and group Sham and group SCI were given equal amount of saline. 1D, 3D, 7d, 14d, 21d and 28d were evaluated by Basso Beattie exercise function after the operation. Scores) and Rivlin oblique plate test were used to evaluate the recovery of nerve function of the spinal cord,.2, and 4W was perfused after injury. The morphological changes of the spinal cord were observed. Dehydration, embedding, and frozen section were observed. After immunofluorescence staining, the images were quantified and the glial fibrillary acidic protein (Glial Fibri), a special marker of astrocytes, was measured. The expression of llary Acidic Protein, GFAP) and the area of glial scar were.3. The morphology of spinal cord and the cavity area of spinal cord were detected by HE staining after injury. Nissl's (Nissl) staining was used to detect the number of Nissl corpuscles and.4. 4 rats were randomly selected from each group after injury, and the fluorescein labeled BDA nerve tracer was injected under stereotyping. After the frozen section of 4W, the number of axon was observed under the fluorescence microscope, and the recovery of spinal nerve conduction pathway was evaluated. [results]1, the BBB score and the Rivlin oblique plate test score were all decreased after the operation. With the increase of postoperative time, the scores of each group were gradually higher than those of the other groups (P0.05), each G was higher than the other groups (P0.05), each G The score of AS in the treatment group was higher than that of the SCI group (P0.05), and the score of SCI+GAS (M) group was higher than that of the other two gastrodin group (P0.05).2, and the GFAP expression of 4W each GAS treatment group and the SCI group increased significantly after the injury, and the expression of the GFAP group was significantly lower than that of the group. (P0.05). The positive staining area of glial scar in each GAS treatment group decreased obviously, and the area of glial scar in SCI+GAS (M) group was obviously lower than that of group SCI, (P0.05).3, 4W in the group after injury and the formation of spinal cord in SCI group, the formation of spinal cord cavity, the spinal cord structure of each GAS treatment group was clearer, the cavity area decreased, and the cavity area of the SCI+GAS (M) group was clear. The number of Nissl corpuscles in group.SCI+GAS (P0.05).SCI+GAS (M) was significantly more than that of group SCI, (P0.05).4. The majority of BDA positive staining nerve fibers in the spinal cord of Sham group after 4 weeks of injury were more than those in the SCI group, and the positive staining fibers in the group were more than the other two groups. On]1, GAS can reduce the GFAP expression level of astrocytes in the injured region of SCI, inhibit astrocyte activation and reduce the formation of.2 in glial scar. GAS can protect the injured region neurons after SCI, reduce the formation of the spinal cord, promote the regeneration of axon, promote the spinal nerve conduction pathway to repair.3, GAS can improve the behavior of rats after SCI. The score could promote the recovery of nerve function, and the optimal concentration range of SCI for treating rats could be around 100mg/Kg.
【學(xué)位授予單位】：昆明醫(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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相關(guān)期刊論文 前5條

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5 Changwei Song;Shiqiang Fang;Gang Lv;Xifan Mei;;Gastrodin promotes the secretion of brain-derived neurotrophic factor in the injured spinal cord[J];Neural Regeneration Research;2013年15期

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本文編號：1984001