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纸质出版:2012
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龙仙萍,石蓓,赵然尊,许官学,崔璨,喻田.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[J].陆军军医大学学报(原第三军医大学学报),2012,34(14):1384-1387.
Long Xianping,Shi Bei,Zhao Ranzun,Xu Guanxue,Cui Can,Yu Tian.Effect of inflammatory cytokine expression in adenovirus vector-mediated human receptor activity modifying protein-1 after carotid artery saccule injury on proliferative intima[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(14):1384-1387.
龙仙萍,石蓓,赵然尊,许官学,崔璨,喻田.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[J].陆军军医大学学报(原第三军医大学学报),2012,34(14):1384-1387. DOI:
Long Xianping,Shi Bei,Zhao Ranzun,Xu Guanxue,Cui Can,Yu Tian.Effect of inflammatory cytokine expression in adenovirus vector-mediated human receptor activity modifying protein-1 after carotid artery saccule injury on proliferative intima[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(14):1384-1387. DOI:
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),P0.05],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(P0.05),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(P0.05)。 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 modifying protein-1(hRAMP1)on expression of inflammatory cytokines in rabbits after carotid artery saccule plastic operation for carotid atherosclerosis. Methods A rabbit atherosclerotic carotid artery stenosis model was established and underwent angioplasty. Thirty-six rabbits were randomly divided into RAMP1 group(n=18)and control group(n=18). On days 7
14 and 28 after they were injected with pAd2-eGFP-RAMP1 or phosphate buffered 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(P0.05). 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
P0.05). However
no significant difference was found in the expression level of CRP on day 28 after injection of adenovirus between the two groups. The hRAMP1 protein was expressed 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 was significantly 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 vs 0.56±0.05
P0.05). Conclusion Exogenous hRAMP1 can regulate the expression of CRP and TNF-α and reduce the restenosis of atherosclerotic carotid artery after angioplasty.
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