中华创伤骨科杂志
中華創傷骨科雜誌
중화창상골과잡지
CHINESE JOURNAL OF ORTHOPAEDIC TRAUMA
2014年
4期
340-344
,共5页
靳宇飞%李富航%毕龙%裴国献
靳宇飛%李富航%畢龍%裴國獻
근우비%리부항%필룡%배국헌
股动脉%交感神经系统%羟多巴胺
股動脈%交感神經繫統%羥多巴胺
고동맥%교감신경계통%간다파알
Femoral artery%Sympathetic nervous system%Oxidopamine
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0.1 mg/mL 6-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28 d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Western blot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14 d取材组几乎不可见交感神经,28 d取材组有少量交感神经.Western blot检测结果表明:对照组、7d取材组、14d取材组及28 d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28 d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.
目的 探討6-羥多巴胺(6-OHDA)在體內去除兔股動脈交感神經的可行性,以期為研究單純血管本身在組織工程骨中有無神經化作用提供理想的動物實驗模型. 方法 分離16隻新西蘭兔右側股動脈,採用不同藥物處理:實驗組採用含0.1 mg/mL 6-OHDA和質量百分比為0.1％抗壞血痠Krebs緩遲液處理,為確定藥物榦預後交感神經的恢複時間,將實驗組再分為7d、14d和28 d取材組(n=4);對照組(n=4)採用僅含0.1％抗壞血痠Krebs緩遲液處理.分彆用囌木精-伊紅(HE)染色、乙醛痠激髮單胺熒光染色檢測股動脈組織變化、交感神經分佈情況,併通過Western blot檢測酪痠羥化酶(TH)和神經肽Y(NPY)蛋白錶達水平的差異. 結果 HE染色結果顯示:與對照組相比,3箇取材組股動脈組織血管壁結構完整,無明顯形態學改變.乙醛痠激髮單胺熒光染色結果顯示:對照組股動脈可見大量交感神經呈網狀和放射狀分佈;7d和14 d取材組幾乎不可見交感神經,28 d取材組有少量交感神經.Western blot檢測結果錶明:對照組、7d取材組、14d取材組及28 d取材組TH/β-actin比值平均分彆為0.70±0.08、0.19±0.08、0.20±0.06、0.33 ±0.06; NPY/β-actin比值平均分彆為0.68±0.14、0.14±0.02、0.17±0.02、0.53±0.10,各取材組的TH和NPY蛋白錶達量較對照組均顯著減少,而28 d取材組的TH和NPY蛋白錶達量較7d、14d取材組明顯增多,差異均有統計學意義(P＜0.05). 結論 體內跼部應用6-OHDA可有效去除股動脈交感神經纖維,是一種可行的血管壁去交感神經方法.本研究建立瞭一種理想的化學去股動脈交感神經的動物模型.
목적 탐토6-간다파알(6-OHDA)재체내거제토고동맥교감신경적가행성,이기위연구단순혈관본신재조직공정골중유무신경화작용제공이상적동물실험모형. 방법 분리16지신서란토우측고동맥,채용불동약물처리:실험조채용함0.1 mg/mL 6-OHDA화질량백분비위0.1％항배혈산Krebs완충액처리,위학정약물간예후교감신경적회복시간,장실험조재분위7d、14d화28 d취재조(n=4);대조조(n=4)채용부함0.1％항배혈산Krebs완충액처리.분별용소목정-이홍(HE)염색、을철산격발단알형광염색검측고동맥조직변화、교감신경분포정황,병통과Western blot검측락산간화매(TH)화신경태Y(NPY)단백표체수평적차이. 결과 HE염색결과현시:여대조조상비,3개취재조고동맥조직혈관벽결구완정,무명현형태학개변.을철산격발단알형광염색결과현시:대조조고동맥가견대량교감신경정망상화방사상분포;7d화14 d취재조궤호불가견교감신경,28 d취재조유소량교감신경.Western blot검측결과표명:대조조、7d취재조、14d취재조급28 d취재조TH/β-actin비치평균분별위0.70±0.08、0.19±0.08、0.20±0.06、0.33 ±0.06; NPY/β-actin비치평균분별위0.68±0.14、0.14±0.02、0.17±0.02、0.53±0.10,각취재조적TH화NPY단백표체량교대조조균현저감소,이28 d취재조적TH화NPY단백표체량교7d、14d취재조명현증다,차이균유통계학의의(P＜0.05). 결론 체내국부응용6-OHDA가유효거제고동맥교감신경섬유,시일충가행적혈관벽거교감신경방법.본연구건립료일충이상적화학거고동맥교감신경적동물모형.
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 subgroup than in the other 2 subgroups (P ＜ 0.05).Conclusions Since local application of 6-OHDA can effectively remove the sympathetic nerve from the femoral artery in a rabbit model,it provides an ideal experimental model with which it is a feasible to remove sympathetic nerves from vascular walls to investigate denervated vascular bundles.
