6-羟多巴胺去除股动脉交感神经的动物模型建立及相关研究  

Local denervation of sympathetic nerve from the femoral artery using 6-hydroxydopamine in a rabbit model

作　　者：靳宇飞[1] 李富航[1] 毕龙[1] 裴国献[1] 

机构地区：[1]第四军医大学西京医院骨科,西安710032

出　　处：《中华创伤骨科杂志》2014年第4期340-344,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（81272011）

摘　　要：目的探讨6-羟多巴胺（6-OHDA）在体内去除兔股动脉交感神经的可行性，以期为研究单纯血管本身在组织工程骨中有无神经化作用提供理想的动物实验模型。方法分离16只新西兰兔右侧股动脉，采用不同药物处理：实验组采用含0．1mg／mL6-OHDA和质量百分比为0．1％抗坏血酸Krebs缓冲液处理．为确定药物干预后交感神经的恢复时间，将实验组再分为7d、14d和28d取材组（n=4）；对照组（n=4）采用仅含0．1％抗坏血酸Krebs缓冲液处理。分别用苏木精．伊红（HE）染色、乙醛酸激发单胺荧光染色检测股动脉组织变化、交感神经分布情况，并通过Westernblot检测酪酸羟化酶（TH）和神经肽Y（NPY）蛋白表达水平的差异。结果HE染色结果显示：与对照组相比，3个取材组股动脉组织血管壁结构完整，无明显形态学改变。乙醛酸激发单胺荧光染色结果显示：对照组股动脉可见大量交感神经呈网状和放射状分布；7d和14d取材组几乎不可见交感神经，28d取材组有少量交感神经。Westernblot检测结果表明：对照组、7d取材组、14d取材组及28d取材组TH／β—actin比值平均分别为0．70±0．08、0．19±0．08、0．20±0．06、0．33±0．06；NPY／β-actin比值平均分别为0．68±0．14、0．14±0．02、0．17±0．02、0．53±0．10，各取材组的TH和NPY蛋白表达量较对照组均显著减少，而28d取材组的TH和NPY蛋白表达量较7d、14d取材组明显增多，差异均有统计学意义（P〈0．05）。结论体内局部应用6-OHDA可有效去除股动脉交感神经纤维，是一种可行的血管壁去交感神经方法。本研究建立了一种理想的化学去股动脉交感神经的动物模型。Objective To explore an effective method to remove sympathetic nerve from the femoral artery in a rabbit model. Methods Femoral arteries from 16 New Zealand rabbits were separated and treated with different drugs. The femoral arteries were processed with Krebs buffer containing 0. 1 mg/mL 6-hydroxydopamine hydrobromide （6-OHDA） and 0. 1% ascorbic acid in the experimental group （ n = 12）, but with Krebs buffer only containing 0. 1% ascorbic acid in the control group （ n = 4） . To investigate the recovery time of sympathetic nerves after drug intervention, the experimental group was divided into 3 subgroups（ n = 4 for each） for sampling at 7, 14 and 28 days respectively. Hematoxylin-eosin （HE） staining was used to detect the structure changes of the femoral artery while glyoxylic acid staining to observe the distribution of sympathetic nerves. The protein expressions of tyrosine hydroxylase （TH） and nerve peptide Y （NPY） were also investigated with western blot. Results HE staining showed that the treatment of 6-OHDA did not damage the normal structure of vascular bundles, compared with the control group. By glyoxylic acid staining, a massive web of sympathetic nerves was distributed in the femoral artery in the control group. In contrast, the sympathetic nerves were almost invisible at 7 and 14 days, and only few of them were observed at 28 days after the treatment of 6-OHDA. The TH/β-actin ratios were 0. 70 ± 0. 08, 0. 19 ± 0. 08, 0. 20 ±0.06 and 0. 33±0. 06 while the NPY/β-actin ratios were 0. 68 ±0. 14, 0. 14 ±0.02, 0. 17 ±0. 02 and 0. 53 ±0. 10 in the control group, 7-day, 14-day and 28-day sampling subgroups, respectively. At all time points, the TH and NPY expressions in all the experimental sub-groups were significantly lower than in the control group （ P 〈 0. 05）, though the expressions were significantly higher in the 28-day sampling sub- group than in the other 2 subgroups （ P 〈 0. 05）. Conclusions Since local application of 6-OHDA can effectively remove

关 键 词：股动脉 交感神经系统 羟多巴胺 

分 类 号：R687.3[医药卫生—骨科学] R-332[医药卫生—外科学]