移植静脉再狭窄进程中Sesn 1蛋白的初步研究

发布时间：2018-06-22 17:39

本文选题：sesn + 1　；参考：《南昌大学》2015年硕士论文

【摘要】：目的:在兔自体移植静脉再狭窄模型的基础上,采用多种研究方法,探究移植静脉再狭窄过程中调控这一过程的关键蛋白,为阐明移植静脉再狭窄的具体机制打下前期基础。方法:将42只健康纯种新西兰大耳白兔随机分为7组,建立兔颈外静脉颈总动脉旁路移植术模型,分别将假手术组正常静脉、术后第1、3、7、14、28、90天移植静脉取出,通过对其进行狭窄程度、PCNA蛋白水平、凋亡等相关检测,确立PCNA蛋白表达差异的时间点后,建立相应时间点的动物模型,并取移植静脉行全基因组表达谱芯片及q RT-PCR等检测。结果:T1组的内膜厚度为4.64±0.7μm;T2组内膜厚度开始有所下降1.46±0.3μm,与T1组比较差异有统计学意义(P0.05);T3组开始内膜厚度持续上升,T3组3.40±0.3μm,T4组内膜厚度达18.2±2.6μm;T1组的中膜+外膜厚度为6.3±0.8μm;术后持续升高。T2组8.3±1.3μm,T3组15.7±1.8μm,T4组45.8±5.1μm,均高于T1组,差异有统计学意义(P0.05)。Western Blot示:T1组PCNA目的条带与内参条带灰度百分比为(40.4±2.6)%;T2组百分比为(36.4±5.2)%,较T1组低,差异有统计学意义(P0.05);随后比值开始上升,T3组百分比为(43.6±4.3)%,较T1组稍高,差异有统计学意义(P0.05);于T4组达到最高峰(78.0±7.3)%,明显高于T1组,差异有统计学意义(P0.05);之后比值缓慢下降。TUNEL法示:T1组中凋亡细胞表达量极低,为(1.46±0.3)%,但在T2组凋亡表达即达高峰(18.06±1.8)%,与T1组比较差异有统计学意义(P0.05);T3组下降较快,为(7.26±0.5)%,与T1组比较差异有统计学意义(P0.05);T4组凋亡降至术后最低水平(2.05±0.6)%,与T1组比较差异有统计学意义(P0.05);之后凋亡维持在较低水平。免疫组化示:T1组PCNA阳性细胞百分比为(6.03±1.0)%;T2组有所下降(5.01±0.9)%,与T1组比较差异有统计学意义(P0.05);T3组开始比值开始增高(11.29±1.4)%,与T1组比较差异有统计学意义(P0.05);于T4组达到最高峰(31.5±3.2)%,与T1组比较差异有统计学意义(P0.05);之后百分比逐渐下降。基因芯片检测分析发现,sesn 1基因在静脉移植术后比正常静脉表达量低,尤其是术后第1天凋亡高峰期时表达量更低。q RT-PCR显示T2组sesn 1的m RNA的表达水平相比于T1组显著下调,差异有统计学意义(P㩳0.05);T4组sesn 1的m RNA的表达水平相比T2组显著上调,差异有统计学意义(P㩳0.05);与基因芯片检测结果中的基因sesn 1的表达趋势相符。结论:sesn 1基因在静脉移植术后比正常静脉表达量低,尤其是术后第1天凋亡高峰期时表达量更低。
[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 卤0.7 渭 m ~ (2) 渭 m ~ (-1), and the intimal thickness of T _ 3 group was 1.46 卤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 卤0.3 渭 m) T _ (4) group was 18.2 卤2.6 渭 m ~ (-1) 渭 m ~ (-1) and the intimal thickness of T _ 3 group was 6.3 卤0.8 渭 m ~ (-1). The continuous increase of T _ 2 group (8.3 卤1.3 渭 m) T _ 3 group (15.7 卤1.8 渭 m) T _ 4 group (45.8 卤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 卤2.6) and (36.4 卤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 卤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 卤7.3), which was significantly higher than that in T1 group (P0.05). (1.46 卤0.3), but the apoptotic expression reached the peak in T2 group (18.06 卤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 卤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 卤1.0) in T _ 2 group and decreased (5.01 卤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 卤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 卤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.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R654

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