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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 investigate the effect of hRAMP1 gene mediated by adenovirus vector on expression of inflammatory cytokines in rabbits with carotid atherosclerosis and balloon angioplasty. Methods Rabbit model of atherosclerosis was established by balloon angioplasty. Randomly divided into RAMP1 group (n = 18) and control group (n = 18), local injection of hRAMP1 gene gland The expression of tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were detected by ELISA method on the 7th, 14th and 28th day after injection. Western blot Local hRAMP1 gene expression was detected by immunohistochemical staining of vascular local TNF-α expression, HE staining for vascular morphology. Results The expression of TNF-α increased at different time points after angioplasty [7 d: (74.13 ± 4.99), 14 d: (93.40 ± 6.69), 28 d: (67.46 ± 6.57)]. (P <0.05). The levels of CRP in RAMP1 group after 7 and 14 days of virus injection [7 d: (64.95 ± 6.77), 14 d: (75.29 ± 4.73), 28 d: (45.08 ± 5.00) : (29.27 ± 1.57), 14 d (9.68 ± 1.60)] compared with the control group [7 d: (43.96 ± 7.88), 14 d: (13.51 ± 1.68)] The expression of hRAMP1 protein was still detected at 28 days after adenovirus injection in rats, and the expression of TNF-α in RAMP1 group was significantly lower than that in control group. HE staining showed that the neointimal area in RAMP1 group [7 days (0.14 ± 0.02), 14 d: (0.39 ± 0.09), 28 d: (0.07 ± 0.18), 14 d: (0.15 ± 0.05), 28 d (0.56 ± 0.05) (P <0.05) .Conclusion Exogenous hRAMP1 gene can regulate the expression of CRP and TNF-α after atherosclerosis and angioplasty in rabbits and inhibit restenosis after angioplasty.