本文選題：sesn + 1　； 參考：《南昌大學(xué)》2015年碩士論文

【摘要】：目的:在兔自體移植靜脈再狹窄模型的基礎(chǔ)上,采用多種研究方法,探究移植靜脈再狹窄過程中調(diào)控這一過程的關(guān)鍵蛋白,為闡明移植靜脈再狹窄的具體機(jī)制打下前期基礎(chǔ)。方法:將42只健康純種新西蘭大耳白兔隨機(jī)分為7組,建立兔頸外靜脈頸總動脈旁路移植術(shù)模型,分別將假手術(shù)組正常靜脈、術(shù)后第1、3、7、14、28、90天移植靜脈取出,通過對其進(jìn)行狹窄程度、PCNA蛋白水平、凋亡等相關(guān)檢測,確立PCNA蛋白表達(dá)差異的時間點后,建立相應(yīng)時間點的動物模型,并取移植靜脈行全基因組表達(dá)譜芯片及q RT-PCR等檢測。結(jié)果:T1組的內(nèi)膜厚度為4.64±0.7μm;T2組內(nèi)膜厚度開始有所下降1.46±0.3μm,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);T3組開始內(nèi)膜厚度持續(xù)上升,T3組3.40±0.3μm,T4組內(nèi)膜厚度達(dá)18.2±2.6μm;T1組的中膜+外膜厚度為6.3±0.8μm;術(shù)后持續(xù)升高。T2組8.3±1.3μm,T3組15.7±1.8μm,T4組45.8±5.1μm,均高于T1組,差異有統(tǒng)計學(xué)意義(P0.05)。Western Blot示:T1組PCNA目的條帶與內(nèi)參條帶灰度百分比為(40.4±2.6)%;T2組百分比為(36.4±5.2)%,較T1組低,差異有統(tǒng)計學(xué)意義(P0.05);隨后比值開始上升,T3組百分比為(43.6±4.3)%,較T1組稍高,差異有統(tǒng)計學(xué)意義(P0.05);于T4組達(dá)到最高峰(78.0±7.3)%,明顯高于T1組,差異有統(tǒng)計學(xué)意義(P0.05);之后比值緩慢下降。TUNEL法示:T1組中凋亡細(xì)胞表達(dá)量極低,為(1.46±0.3)%,但在T2組凋亡表達(dá)即達(dá)高峰(18.06±1.8)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);T3組下降較快,為(7.26±0.5)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);T4組凋亡降至術(shù)后最低水平(2.05±0.6)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);之后凋亡維持在較低水平。免疫組化示:T1組PCNA陽性細(xì)胞百分比為(6.03±1.0)%;T2組有所下降(5.01±0.9)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);T3組開始比值開始增高(11.29±1.4)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);于T4組達(dá)到最高峰(31.5±3.2)%,與T1組比較差異有統(tǒng)計學(xué)意義(P0.05);之后百分比逐漸下降�；蛐酒瑱z測分析發(fā)現(xiàn),sesn 1基因在靜脈移植術(shù)后比正常靜脈表達(dá)量低,尤其是術(shù)后第1天凋亡高峰期時表達(dá)量更低。q RT-PCR顯示T2組sesn 1的m RNA的表達(dá)水平相比于T1組顯著下調(diào),差異有統(tǒng)計學(xué)意義(P㩳0.05);T4組sesn 1的m RNA的表達(dá)水平相比T2組顯著上調(diào),差異有統(tǒng)計學(xué)意義(P㩳0.05);與基因芯片檢測結(jié)果中的基因sesn 1的表達(dá)趨勢相符。結(jié)論:sesn 1基因在靜脈移植術(shù)后比正常靜脈表達(dá)量低,尤其是術(shù)后第1天凋亡高峰期時表達(dá)量更低。
[Abstract]:Objective: on the basis of the autograft vein restenosis model in rabbits, the key proteins involved in the process of vein graft restenosis were investigated by various research methods, which laid a foundation for elucidating the mechanism of vein graft restenosis. Methods: Forty-two healthy New Zealand white rabbits were randomly divided into 7 groups. The common jugular artery bypass grafting model was established in rabbits. The normal veins in the sham-operated group were removed on the 1st day after operation. By detecting the level of PCNA protein and apoptosis, we established the time point of the difference of PCNA protein expression, established the animal model of the corresponding time point, and took the vein grafts to detect the whole genome expression microarray and Q RT-PCR. Results the intimal thickness of T _ 1 group was 4.64 鹵0.7 渭 m ~ (2) 渭 m ~ (-1), and the intimal thickness of T _ 3 group was 1.46 鹵0.3 渭 m ~ (3) 渭 m, which was significantly higher than that of T _ (1) group (P < 0.05). The intimal thickness of T _ 3 group (3.40 鹵0.3 渭 m) T _ (4) group was 18.2 鹵2.6 渭 m ~ (-1) 渭 m ~ (-1) and the intimal thickness of T _ 3 group was 6.3 鹵0.8 渭 m ~ (-1). The continuous increase of T _ 2 group (8.3 鹵1.3 渭 m) T _ 3 group (15.7 鹵1.8 渭 m) T _ 4 group (45.8 鹵5.1 渭 m) was higher than that of T _ 1 group (P < 0.05). The difference was statistically significant (P0.05). Western Blot showed that the grayscale percentage of PCNA target band and internal reference band was (40.4 鹵2.6) and (36.4 鹵5.2) in the W T1 group, which was lower than that in the T1 group (P0.05), then the percentage of T _ 3 group was (43.6 鹵4.3), which was slightly higher than that in the T1 group. The difference was statistically significant (P0.05), and reached the peak in T4 group (78.0 鹵7.3), which was significantly higher than that in T1 group (P0.05). (1.46 鹵0.3), but the apoptotic expression reached the peak in T2 group (18.06 鹵1.8). There was a significant difference between T1 group and T1 group (P0.05). Compared with T1 group, apoptosis in T4 group decreased to the lowest level (2.05 鹵0.6), and there was significant difference compared with T1 group (P0.05). Immunohistochemical staining showed that the percentage of PCNA positive cells decreased (6.03 鹵1.0) in T _ 2 group and decreased (5.01 鹵0.9) in T _ 2 group, which was significantly higher than that in T _ 1 group (P 0.05). The ratio of PCNA positive cells in T _ 3 group began to increase (11.29 鹵1.4), which was significantly higher than that in T _ 1 group (P 0.05), and reached the peak in T _ 4 group (31.5 鹵3.2). Compared with T1 group, the difference was statistically significant (P0.05), and then the percentage gradually decreased. The results of microarray analysis showed that the expression of sesn1 gene was lower than that of normal vein after vein transplantation, especially at the peak of apoptosis on the first day after transplantation. Q RT-PCR showed that the mRNA expression of sesn 1 in T2 group was significantly lower than that in T1 group. The mRNA expression level of sesn 1 in T4 group was significantly higher than that in T2 group (P0. 05), which was consistent with the trend of gene sesn 1 expression in gene microarray. Conclusion the expression of 1% sesn1 gene was lower after vein transplantation than that in normal vein, especially at the peak of apoptosis on the first day after operation.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R654

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 萬力;王文俊;曹園平;王群;劉季春;;纖維蛋白膠外支架轉(zhuǎn)染PCNA基因反義寡核苷酸預(yù)防移植靜脈再狹窄[J];中國組織工程研究與臨床康復(fù);2011年38期

2 魏翔;吳黎;李軍;周雁榮;張毅;徐利軍;潘鐵成;朱學(xué)海;;靜脈移植血管的線粒體氧化損傷機(jī)制研究[J];中華實驗外科雜志;2010年05期

3 王明松;鄒良建;黃盛東;徐志云;鄒榕江;;自體移植靜脈內(nèi)皮細(xì)胞的缺血安全時限[J];中國組織工程研究與臨床康復(fù);2007年08期

，

本文編號：2053624