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西罗莫司抑制肾移植后AGE诱导动脉粥样硬化形成的作用及其机制

Effect of rapamycin on formation of atherosclerosis induced by AGEs in renal transplantation recipients and mechanism

摘要:

目的 探讨肾移植术后西罗莫司抑制晚期糖基化终末产物(AGE)诱导的动脉粥样硬化(AS)形成的作用及其机制.方法 纳入5例肾移植后确诊AS形成的受者,受者在发生AS前接受过移植肾穿刺活检,确诊AS后则使用西罗莫司抗排斥反应治疗1年以上,观察使用西罗莫司前后移植肾血管组织的结构变化.体外分离和培养SD大鼠主动脉平滑肌细胞(VSMC),并以免疫荧光染色法进行鉴定;以第5代VSMC为体外实验对象,取第5代VSMC,将细胞分为西罗莫司组(经西罗莫司培养)、AGE组(经AGE培养)、实验组(经西罗莫司和AGE共培养)及对照组(经牛血清蛋白培养),收集各组VSMC,采用蛋白质印迹法检测VSMC上α平滑肌肌动蛋白(α-SMA)、骨桥蛋白(OPN)和增殖细胞核抗原(PCNA)的表达.结果 肾移植后AS形成受者的血管中膜明显增厚,而使用西罗莫司处理(13.0±1.8)个月后,中膜厚度则显著降低.体外实验表明,正常SD大鼠第2、5代VSMC中α-SMA均高度表达;经AGE作用后可诱导VSMC中α-SMA表达量明显减少(P<0.05);而OPN和PCNA的表达明显升高(P<0.05).此外,与AGE组相比,实验组VSMC中PCNA和OPN的表达明显下降,而α-SMA表达量明显上升,差异均有统计学意义(P<0.05).结论 西罗莫司可能通过抑制AGE诱导的血管平滑肌细胞增殖及其表型的转分化进而抑制AGE引起的肾移植后AS的进展.

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abstracts:

Objective +o investigate the inhibitory effect of sirolimus on formation of atherosclerosis (AS) induced by advanced glycation end products (AGE) in kidney transplantation recipients and mechanism.Method Five cases of definite diagnosis of AS were included.Structural changes of renal transplant vascular tissues were observed before and after the application of sirolimus for at least over 1 year once the definite diagnosis of AS was made by transplant kidney aspiration biopsy.SD rat aortic smooth muscle cells (VSMC) were isolated and cultured in vitro and identified by immunofluorescence staining.Sirolimus group (sirolimus culture),AGE group (AGE culture),experimental group (sirolimus and AGE co-culture) and control group (bovine serum albumin culture) were established.VSMCs of the 5th generation from each group were collected and the expression levels of α-smooth muscle actin (α-SMA),osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) were detected by Western blotting.Result Significant thickness was found in the media layer of vessels in allograft kidney from recipients with AS comparing with significant attenuation after the administration of sirolimus for 13.0 + 1.8 months.+he in vitro experiments showed that α-SMA was overexpressed in both 2 nd and 5 th generation VSMCs of normal SD rats; in contrast,low expression of α-SMA and high expression of OPN and PCNA were detected after the exposure of AGE (P< 0.05).Moreover,as compared with AGE group,PCNA and OPN were significantly decreased,and α-SMA was significantly increased in experimental group (P<0.05).Conclusion Sirolimus can inhibit the progression of post-transplant AS induced by AGE possibly through suppressing the proliferation of VSMCs and transdifferentiation of their phenotypes.

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作者: 刘绪忠 [1] 刘超 [1] 汪俊 [1] 韩志坚 [1] 陶俊 [1] 鲁佩 [1] 殷长军 [1] 谭若芸 [1] 顾民 [1]
栏目名称: 临床研究
DOI: 10.3760/cma.j.issn.0254-1785.2014.04.005
发布时间: 2014-05-28
基金项目:
国家自然科学基金青年基金项目 江苏省“333高层次人才培养工程”项目 江苏省“六大人才高峰”资助项目 江苏省“科教兴卫”工程医学重点人才培养资助项目 江苏省卫生厅面上项目
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