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  本文選題：早期生長反應因子-1 + 基質金屬蛋白酶-9��； 參考：《中國醫(yī)科大學》2010年碩士論文

【摘要】： 目的 通過轉染針對早期生長反應因子-1(early growth response factor-1,Egr-1)的誘騙寡核苷酸(Egr-1 decoy ODNs)到大鼠動脈血管平滑肌細胞中,檢測各組早期生長反應因子-1、基質金屬蛋白酶-9(Matrix metalloproteinases-9,MMP-9)表達變化及測量各組平滑肌細胞(vascular smooth muscle cell, VSMC)的遷移,探討針對早期生長反應因子-1的誘騙寡核苷酸對體外培養(yǎng)的大鼠平滑肌細胞遷移的影響和作用機制。 方法 通過組織塊貼壁法進行原代大鼠動脈血管平滑肌細胞培養(yǎng)、傳代培養(yǎng)和鑒定,設計合成Egr-1誘騙、誘騙對照寡核苷酸。轉染人工合成的針對Egr-1的誘騙寡核苷酸(Egr-1 decoy ODN),將培養(yǎng)的平滑肌細胞分為對照組(正常培養(yǎng)細胞)、誘騙組(轉染Egr-1的誘騙寡核苷酸)、誘騙對照組(轉染Egr-1誘騙雜碼寡核苷酸)。免疫組織化學染色法檢測各組不同時間Egr-1及基質金屬蛋白酶-9(MMP-9)蛋白的表達,進行圖像分析。對各組采用劃痕法測定平滑肌細胞的遷移距離。所有數(shù)據(jù)均采用SPSS13.0統(tǒng)計軟件進行統(tǒng)計分析,計量資料以均值±標準差(x±S)表示,各組間比較應用單因素方差分析,P0.05有統(tǒng)計學意義。 結果 成功進行原代VSMC培養(yǎng),經(jīng)細胞鑒定培養(yǎng)細胞為高純度的血管平滑肌細胞。轉染Egr-1誘騙寡核苷酸后,用免疫組織化學法分別觀察各組不同時間點的Egr-1和MMP-9蛋白的表達情況。實驗發(fā)現(xiàn)：Egr-1誘騙寡核苷酸轉染后誘騙組的遷移距離明顯低于誘騙對照組和對照組(P0.05),誘騙對照組和對照組沒有明顯的差異。同時發(fā)現(xiàn)Egr-1誘騙寡核苷酸轉染后Egr-1及MMP-9蛋白在誘騙組的表達較低,與誘騙對照組和對照組比較具有統(tǒng)計學差異。 結論 本實驗通過體外轉染Egr-1的誘騙寡核苷酸在一定程度上能夠通過抑制Egr-1蛋白的表達,抑制其下游MMP-9基因的表達,減少血清誘導的平滑肌細胞的遷移,從而達到抑制細胞遷移的目的。
[Abstract]:Objective to transfect Egr-1 decoy ODNs targeting early growth response factor-1 (Egr-1) into rat vascular smooth muscle cells. The expression of early growth response factor-1, matrix metalloproteinases-9 (MMP-9) and the migration of smooth muscle cells (vascular smooth muscle cell, VSMC) in each group were measured. To investigate the effect and mechanism of decoy oligonucleotides targeting early growth response factor-1 on the migration of rat smooth muscle cells in vitro. Methods the primary vascular smooth muscle cells of rat artery were cultured, cultured and identified by tissue mass adherence. Egr-1 decoy was designed and synthesized, and the control oligonucleotides were decoded. The cultured smooth muscle cells were divided into control group (normal cultured cells), decoy group (Egr-1 decoy oligonucleotide) and control group (transfected Egr-1 decoy oligonucleotide). The expression of Egr-1 and matrix metalloproteinase-9 (MMP-9) were detected by immunohistochemical staining. The migration distance of smooth muscle cells was measured by scratch method. All the data were analyzed by SPSS 13.0 statistical software. The measurement data were expressed as mean 鹵standard deviation (x 鹵S). Results the primary VSMC was successfully cultured and the cultured cells were identified as high purity vascular smooth muscle cells. The expression of Egr-1 and MMP-9 were observed by immunohistochemical method after transfection of Egr-1 decoded oligonucleotides. It was found that the migration distance of the decoy group was significantly lower than that of the decoy control group and the control group after transfection (P0.05), but there was no significant difference between the decoy control group and the control group. It was also found that the expression of Egr-1 and MMP-9 protein was lower in the decoy group after transfection with Egr-1 decoy oligodeoxynucleotides, which was significantly different from that in the decoy control group and the control group. Conclusion Egr-1 induced oligonucleotides can inhibit the expression of MMP-9 gene in the downstream of Egr-1 and reduce the migration of smooth muscle cells induced by serum to a certain extent by inhibiting the expression of Egr-1 protein and the expression of MMP-9 gene in the downstream of Egr-1. In order to achieve the purpose of inhibiting cell migration.
【學位授予單位】：中國醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R363

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本文編號：2108725