本文選題：下肢動(dòng)脈硬化閉塞癥 + 支架內(nèi)再狹窄�。� 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:觀察RCAN1、鈣調(diào)神經(jīng)磷酸酶A(CnA)在下肢動(dòng)脈硬化支架術(shù)后支架內(nèi)再狹窄組織中的表達(dá),探討其與再狹窄的作用及相關(guān)性。方法:收集2013年9月至2016年6月重慶醫(yī)科大學(xué)附屬第一醫(yī)院血管外科中心下肢閉塞性動(dòng)脈硬化股淺動(dòng)支架術(shù)后再狹窄截肢患者血管標(biāo)本15例,取支架段、支架近端,支架遠(yuǎn)端血管組織,采用HE及Masson染色觀察血管組織形態(tài)學(xué)變化;Real-time PCR檢測(cè)RCAN1mRNA,CN mRNA在各組中的表達(dá);Western blotting檢測(cè)RCAN1、CnA、增殖細(xì)胞核抗原(PCNA)在支架段、支架近端,支架遠(yuǎn)端血管組織的表達(dá);免疫組化檢測(cè)RCAN1、CnA、a-SMA蛋白的表達(dá),免疫熒光檢測(cè)RCAN1、CnA蛋白分布;免疫共沉淀檢測(cè)RCAN與CnA蛋白之間的互相作用。結(jié)果:形態(tài)學(xué)觀察示:支架術(shù)后3-6月支架狹窄段血管組織形態(tài)變化明顯,管腔閉塞,支架腔內(nèi)長(zhǎng)滿新生組織;支架近端血管內(nèi)膜組織顯著增厚,血管平滑肌細(xì)胞排列紊亂;支架遠(yuǎn)端組織血管內(nèi)膜稍增厚,血管平滑肌組織排列整齊。Western blotting及Real-time PCR結(jié)果顯示RCAN1在支架遠(yuǎn)端組織的表達(dá)水平顯著高于支架近端組織和支架狹窄段組織的表達(dá)水平(P0.05),RCAN1在支架近端組織的表達(dá)水平顯著高于支架狹窄段組織(P0.05);CnA在支架狹窄段組織的表達(dá)顯著高于支架近端組織(P0.05),其在支架狹窄段組織的表達(dá)顯著高于支架近端組織(P0.05)。PCNA在狹窄組織中的表達(dá)顯著高于支架近端及支架遠(yuǎn)端組織。免疫組化結(jié)果示a-SMA在支架狹窄段新生組織中表達(dá),RCAN1和CnA主要在細(xì)胞質(zhì)中表達(dá);免疫熒光結(jié)果顯示RCAN1和CnA與a-SMA共表達(dá)。免疫共沉淀示RCAN1與CnA蛋白在血管組織中存在相互作用。結(jié)論:RCAN1蛋白、CnA在人體動(dòng)脈支架術(shù)后血管組織中異常表達(dá);RCAN1的表達(dá)水平降低與CnA的表達(dá)水平增高可能與支架內(nèi)再狹窄發(fā)生有關(guān);RCAN1-CN信號(hào)通路可能通過(guò)調(diào)節(jié)VSMC的增殖進(jìn)而參與再狹窄的病理過(guò)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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R654.4

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本文編號(hào)：1965135