腺病毒载体介导hRAMP1调节兔颈动脉球囊损伤后炎症细胞因子表达对增生内膜的影响  

Effect of inflammatory cytokine expression in adenovirus vector-mediated human receptor activity modifying protein-1 after carotid artery saccule injury on proliferative intima

作　　者：龙仙萍[1] 石蓓[1] 赵然尊[1] 许官学[1] 崔璨[1] 喻田[2] 

机构地区：[1]遵义医学院第一附属医院心血管内科,贵州遵义563003 [2]遵义医学院医学与分子生物学研究中心,贵州遵义563003

出　　处：《第三军医大学学报》2012年第14期1384-1387,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81060014);麻醉与器官保护基础研究重点实验室专项资金(黔科合技工字[2011]4006)~~

摘　　要：目的探讨腺病毒载体介导人受体活性修饰蛋白-1(receptor activity modifying protein-1,hRAMP1)基因对兔颈动脉粥样硬化并球囊成形术后炎性细胞因子表达的影响。方法建立兔动脉粥样硬化狭窄模型并行球囊损伤血管(简称血管成形术),随机抽样分为RAMP1组(n=18)和对照组(n=18),经球囊局部注射携带hRAMP1基因腺病毒载体(pAd2-GFP-RAMP1)或PBS,于注射后7、14 d和28 d,应用ELISA法检测肿瘤坏死因子-α(TNF-α)和C-反应蛋白(CRP)表达水平;Western blot检测局部hRAMP1目的基因表达;免疫组织化学染色测定血管局部TNF-α表达,HE染色检测血管形态学。结果血管成形术后不同时间点TNF-α表达增加[7 d:(74.13±4.99),14 d:(93.40±6.69),28 d:(67.46±6.57)],外源hRAMP1注射后TNF-α表达下降[7 d:(64.95±6.77),14 d:(75.29±4.73),28 d:(45.08±5.00),P<0.05],病毒注射后7 d和14 d RAMP1组CRP水平[7 d:(29.27±1.57),14 d:(9.68±1.60)]与对照组比较[7 d:(43.96±7.88),14 d:(13.51±1.68)]显著下降(P<0.05),28 d 2组间无差异性;腺病毒注射后28 d损伤血管局部仍检测到hRAMP1蛋白表达,同时RAMP1组局部TNF-α表达与对照组比较明显下降,HE染色显示:RAMP1组新生内膜面积[7 d:(0.07±0.18),14 d:(0.15±0.05),28 d:(0.35±0.05)]与对照组[7 d:(0.14±0.02),14 d:(0.39±0.09),28 d:(0.56±0.05]比较明显降低(P<0.05)。结论外源hRAMP1基因调节兔动脉粥样硬化并血管成形术后CRP和TNF-α的表达,抑制血管成形术后再狭窄。Objective To study the effect of adenovirus vector-mediated human receptor activity modi- fying protein-1 （ hRAMP1 ） on expression of inflammatory cytokines in rabbits after carotid artery saceule plastic operation for carotid atherosclerosis. Methods A rabbit atheroselerotic carotid artery stenosis model was estab- lished and underwent angioplasty. Thirty-six rabbits were randomly divided into RAMP1 group（ n = 18 ）and con- trol group（n = 18）. On days 7, 14 and 28 after they were injected with pAd2-eGFP-RAMP1 or phosphate buff- ered saline（PBS） , serum levels of TNF-α and CRP were measured by ELISA. Expression of hRAMP1 target gene in carotid artery was detected by Western blotting and expression of TNF-α in carotid artery was detected with immunohistochemical staining. Morphology of carotid artery was observed with HE staining. Results The expression level of TNF-α was 74.13 ± 4.99, 93.40 ± 6.69 and 67.46± 6.57, respectively, on days 7, 14 and 28 after angioplasty, and 64.95± 6.77, 75.29±4.73 and 45.08 ± 5.00, respectively, on days 7, 14 and 28 after injection of exogeneous hRAMP1 （ P 〈 0. 05 ）. The expression level of CRP was significantly lower in RAMP1 group than in control group on days 7 and 14 after injection of adenovirus （29.27 ± 1.57 vs 43.96 ±7.88, 9.68±1.60 vs 13.51± 1.68, P 〈 0. 05）. However, no significant difference was found in the expres- sion level of CRP on day 28 after injection of adenovirus between the two groups. The hRAMP1 protein was ex- pressed in local injured blood vessels on day 28 after injection of adenovirus and the TNF-α expression level was significantly lower in hRAMP1 group than in control group. HE staining showed that the neointimal area wassignificantly smaller in hRAMP1 group than in control group on days 7, 14 and 28 after injection of adenovirus （0.07±0.18 vs 0. 14 ±0.02, 0.15±0.05 vs 0.39 ±0.09, 0.35±0.05 vs0.56±0.05, P 〈0.05）. Conclusion Exogenous hRAMP1 can regulate the expression of CRP and TNF-α and reduce the re

关 键 词：人受体活性修饰蛋白-1 炎症细胞因子 血管成形术 增生内膜 

分 类 号：R392-33[医药卫生—免疫学]