【摘要】：【目的】觀察并分析大鼠急性脊髓損傷后microRNA-223和RhoB基因表達變化及二者間的關(guān)系,探索治療脊髓損傷潛在的靶點,提供理論基礎(chǔ)。【方法】采用改良Allen,s法建立大鼠急性脊髓損傷模型;將108只大鼠隨機分為假手術(shù)組(A組、對照組,18只)、脊髓損傷組(B組、實驗組),其中B組分為SCI后6h、12h、24h、3d、7d組,每組各18只;各組分別于相應(yīng)時相行BBB評分評價神經(jīng)功能、HE染色觀察組織形態(tài)學變化、實時熒光定量PCR檢測miRNA-223和RhoB基因表達、原位雜交檢測miRNA-223和RhoB表達的定位關(guān)系。【結(jié)果】1、BBB評分:SCI后各時間組較無損傷對照組評分明顯降低,差異均有統(tǒng)計學意義。2、HE染色:正常脊髓組織神經(jīng)元細胞形態(tài)結(jié)構(gòu)正常;損傷后6h組脊髓組織廣泛出現(xiàn)水腫;12-24h后發(fā)展為壞死組織,灰質(zhì)出現(xiàn)炎性細胞浸潤;損傷后24h-7d,毛細血管通透性增加,組織液滲出,神經(jīng)元細胞廣泛溶解、壞死,大量炎癥細胞浸潤。3、實時熒光定量PCR:SCI后各實驗組的脊髓組織中miRNA-223表達均升高,與無損傷對照組相比,差異均有統(tǒng)計學意義(P0.05);SCI后各組RhoB表達逐漸增高,24h時達到高峰,隨之下降,至7d時表達仍高于對照組,差異有統(tǒng)計學意義(P0.05)。急性脊髓損傷后miRNA-223、RhoB兩種基因的表達呈顯著正相關(guān)(r=0.89)。4、原位雜交:miRNA-223主要表達于SCI后6h、12h、24h組,而RhoB主要表達于SCI后6h、12h、24h、3d組。【結(jié)論】1、脊髓損傷后miRNA-223基因表達顯著升高,而RhoB的表達同步明顯上調(diào),二者呈顯著正相關(guān)。2、RhoB基因表達可能受miRNA-223調(diào)控。
[Abstract]:[objective] to observe and analyze the changes of microRNA-223 and RhoB gene expression and the relationship between microRNA-223 and RhoB gene after acute spinal cord injury in rats, and to explore the potential targets for the treatment of spinal cord injury. [methods] the model of acute spinal cord injury was established by modified Alleners method, 108 rats were randomly divided into sham operation group (group A, control group, n = 18) and spinal cord injury group (group B, experimental group). 18 rats in each group were evaluated with BBB score at the same time, and the histomorphologic changes were observed by HE staining, and the expression of miRNA-223 and RhoB genes were detected by real-time fluorescence quantitative PCR. In situ hybridization was used to detect the localization of miRNA-223 and RhoB expression. [results] 1BBB score was significantly lower in each time group than that in the control group (P < 0.01). The difference was statistically significant. 2 he staining showed that the morphology and structure of neurons in normal spinal cord were normal. 6 h after injury, the spinal cord tissue developed into necrotic tissue and inflammatory cell infiltration in gray matter after 12 to 24 hours of injury, capillary permeability increased, tissue fluid exudated, neuronal cells dissolved and necrotized widely at 24 h after injury. The expression of miRNA-223 in the spinal cord of each experimental group was increased after real-time fluorescence quantitative PCR: sci infiltration. Compared with the control group, the difference was statistically significant (P0.05) after sci, the expression of RhoB reached its peak at 24 hours after sci, and then decreased. At 7 days, the expression was still higher than the control group, the difference was statistically significant (P0.05). After acute spinal cord injury, the expression of miRNA-223hRhoB gene was positively correlated (r: 0.89) .4. In situ hybridization, miRNA-223 was mainly expressed in 24 h group after sci, while RhoB was mainly expressed in group 6 h after sci and 24 h after sci. [conclusion] 1, miRNA-223 gene expression was significantly increased after spinal cord injury. The expression of RhoB was significantly up-regulated, and the expression of RhoB gene was positively correlated with that of miRNA-223.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.2

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相關(guān)期刊論文 前1條

1 汪建良;余勤;白月雙;;脊髓損傷動物模型研究現(xiàn)狀[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2009年02期

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本文編號：2119936