大鼠损伤脊髓中microRNA-223的表达及与RhoB的相关性研究
发布时间：2018-07-13 15:45
【摘要】：【目的】观察并分析大鼠急性脊髓损伤后microRNA-223和RhoB基因表达变化及二者间的关系,探索治疗脊髓损伤潜在的靶点,提供理论基础。【方法】采用改良Allen,s法建立大鼠急性脊髓损伤模型;将108只大鼠随机分为假手术组(A组、对照组,18只)、脊髓损伤组(B组、实验组),其中B组分为SCI后6h、12h、24h、3d、7d组,每组各18只;各组分别于相应时相行BBB评分评价神经功能、HE染色观察组织形态学变化、实时荧光定量PCR检测miRNA-223和RhoB基因表达、原位杂交检测miRNA-223和RhoB表达的定位关系。【结果】1、BBB评分:SCI后各时间组较无损伤对照组评分明显降低,差异均有统计学意义。2、HE染色:正常脊髓组织神经元细胞形态结构正常;损伤后6h组脊髓组织广泛出现水肿;12-24h后发展为坏死组织,灰质出现炎性细胞浸润;损伤后24h-7d,毛细血管通透性增加,组织液渗出,神经元细胞广泛溶解、坏死,大量炎症细胞浸润。3、实时荧光定量PCR:SCI后各实验组的脊髓组织中miRNA-223表达均升高,与无损伤对照组相比,差异均有统计学意义(P0.05);SCI后各组RhoB表达逐渐增高,24h时达到高峰,随之下降,至7d时表达仍高于对照组,差异有统计学意义(P0.05)。急性脊髓损伤后miRNA-223、RhoB两种基因的表达呈显著正相关(r=0.89)。4、原位杂交:miRNA-223主要表达于SCI后6h、12h、24h组,而RhoB主要表达于SCI后6h、12h、24h、3d组。【结论】1、脊髓损伤后miRNA-223基因表达显著升高,而RhoB的表达同步明显上调,二者呈显著正相关。2、RhoB基因表达可能受miRNA-223调控。
[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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
本文编号：2119936
[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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
【参考文献】
相关期刊论文 前1条
1 汪建良;余勤;白月双;;脊髓损伤动物模型研究现状[J];现代中西医结合杂志;2009年02期
，本文编号：2119936
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