RCAN1及CnA在下肢动脉硬化闭塞支架术后再狭窄组织中表达及意义

发布时间：2018-06-01 18:03

本文选题：下肢动脉硬化闭塞症 + 支架内再狭窄　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:观察RCAN1、钙调神经磷酸酶A(CnA)在下肢动脉硬化支架术后支架内再狭窄组织中的表达,探讨其与再狭窄的作用及相关性。方法:收集2013年9月至2016年6月重庆医科大学附属第一医院血管外科中心下肢闭塞性动脉硬化股浅动支架术后再狭窄截肢患者血管标本15例,取支架段、支架近端,支架远端血管组织,采用HE及Masson染色观察血管组织形态学变化;Real-time PCR检测RCAN1mRNA,CN mRNA在各组中的表达;Western blotting检测RCAN1、CnA、增殖细胞核抗原(PCNA)在支架段、支架近端,支架远端血管组织的表达;免疫组化检测RCAN1、CnA、a-SMA蛋白的表达,免疫荧光检测RCAN1、CnA蛋白分布;免疫共沉淀检测RCAN与CnA蛋白之间的互相作用。结果:形态学观察示:支架术后3-6月支架狭窄段血管组织形态变化明显,管腔闭塞,支架腔内长满新生组织;支架近端血管内膜组织显著增厚,血管平滑肌细胞排列紊乱;支架远端组织血管内膜稍增厚,血管平滑肌组织排列整齐。Western blotting及Real-time PCR结果显示RCAN1在支架远端组织的表达水平显著高于支架近端组织和支架狭窄段组织的表达水平(P0.05),RCAN1在支架近端组织的表达水平显著高于支架狭窄段组织(P0.05);CnA在支架狭窄段组织的表达显著高于支架近端组织(P0.05),其在支架狭窄段组织的表达显著高于支架近端组织(P0.05)。PCNA在狭窄组织中的表达显著高于支架近端及支架远端组织。免疫组化结果示a-SMA在支架狭窄段新生组织中表达,RCAN1和CnA主要在细胞质中表达;免疫荧光结果显示RCAN1和CnA与a-SMA共表达。免疫共沉淀示RCAN1与CnA蛋白在血管组织中存在相互作用。结论:RCAN1蛋白、CnA在人体动脉支架术后血管组织中异常表达;RCAN1的表达水平降低与CnA的表达水平增高可能与支架内再狭窄发生有关;RCAN1-CN信号通路可能通过调节VSMC的增殖进而参与再狭窄的病理过程。
[Abstract]:Aim: to investigate the expression of RCAN1, calmodulin C _ (nA) in restenosis tissue after lower extremity arteriosclerotic stenting, and to explore its role and correlation with restenosis. Methods: from September 2013 to June 2016, 15 patients with restenosis after shallow-femoral stenting were collected from vascular surgery center of the first affiliated Hospital of Chongqing Medical University. The expression of RCAN1mRNA-CN mRNA and proliferating cell nuclear antigen (PCNA) in the stents, proximal stents and distal stents were detected by HE and Masson staining. The expression of RCAN1mRNA-CN mRNA in each group was detected by real-time PCR. The proliferating cell nuclear antigen (PCNA) was detected in the stent segment, the proximal end of the stent and the distal vascular tissue of the stent. Immunohistochemistry was used to detect the expression of RCAN1CnAfa-SMA protein, immunofluorescence was used to detect the distribution of RCAN1CnA protein, and the interaction between RCAN and CnA protein was detected by immunoprecipitation. Results: morphological observation showed that the morphology of vascular tissue in the stenotic segment of stent was obvious in 3-6 months after stenting, the lumen was occluded, the stent cavity was filled with new tissue, the intimal tissue of proximal end of stent was significantly thickened, and the vascular smooth muscle cells were arranged in disorder. The intima of the distal tissue of the stent was slightly thicker. The results of neatly arranged vascular smooth muscle tissue. Western blotting and Real-time PCR showed that the expression level of RCAN1 in distal stents was significantly higher than that in proximal stents and stenotic stents. The expression of P0.05CnA in the stenotic stents was significantly higher than that in the proximal stents, and the expression of PCNA in the stenotic stents was significantly higher than that in the proximal stents. Proximal and distal tissue of scaffold. Immunohistochemical results showed that a-SMA was mainly expressed in cytoplasm and RCAN1 and CnA co-expressed with a-SMA in the neoplasm of stenosed stents. Immunoprecipitation showed that RCAN1 and CnA protein interacted with each other in vascular tissues. Conclusion the decreased expression level of RCAN1 and the increased expression of CnA may be related to the development of restenosis in the stents, which may be related to the RCAN1-CN signaling pathway by regulating the proliferation of VSMC. Conclusion the abnormal expression of RCAN1 protein in vascular tissue after stenting may be related to the decrease of RCAN1 expression level and the increase of CnA expression level. And participate in the pathological process of restenosis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R654.4

【参考文献】