中国康复理论与实践
中國康複理論與實踐
중국강복이론여실천
CHINESE JOURNAL OF REHABILITATION THEORY & PRACTICE
2014年
8期
701-708
,共8页
郭杨%马勇%潘娅岚%成吉华%黄桂成
郭楊%馬勇%潘婭嵐%成吉華%黃桂成
곽양%마용%반아람%성길화%황계성
脊髓损伤%脊髓康%轴突再生%微环境%脑源性神经营养因子%大鼠
脊髓損傷%脊髓康%軸突再生%微環境%腦源性神經營養因子%大鼠
척수손상%척수강%축돌재생%미배경%뇌원성신경영양인자%대서
spinal cord injury%Jisuikang%axonal regeneration%microenvironment%brain-derived neurotrophic factor%rats
目的:探讨脊髓康对脊髓损伤(SCI)后神经功能恢复以及脑源性神经营养因子(BDNF)蛋白和mRNA表达的影响。方法144只雌性Sprague-Dawley大鼠,体质量180~220 g,随机抽取24只作为假手术组,仅咬除T9~T11段椎板、棘突。其余采用改良Allen法成功建立SCI动物模型后,随机分为5组,即模型组、醋酸泼尼松组及脊髓康高、中、低剂量组,每组24只。脊髓康高、中、低剂量组分别按生药剂量50 g/(kg?d)、25 g/(kg?d)、12.5 g/(kg?d)灌胃;醋酸泼尼松组以醋酸泼尼松0.06 g/(kg?d)灌胃;假手术组与模型组均以同体积生理盐水灌胃。分别于术后24 h、3 d、7 d、14 d,进行Basso-Beattie-Bresnahan(BBB)评分、斜板试验,干预后3 d、7 d、14 d处死大鼠取脊髓样本,采用免疫组化、Western blotting、荧光定量PCR法检测脊髓损伤区BDNF蛋白和mRNA的表达情况。结果术后24 h假手术组大鼠BBB评分、斜板试验角度明显高于其他各组(P<0.01)。术后3~14 d脊髓康中剂量组和醋酸泼尼松组BBB评分均高于模型组(P<0.05),且术后14 d脊髓康中剂量组BBB评分明显高于其他各组(P<0.01)。免疫组化和Western blotting显示,不同时间点醋酸泼尼松组和脊髓康中剂量组BDNF蛋白表达水平均明显高于模型组(P<0.01),二者具有等效性(P>0.05)。荧光定量PCR显示,醋酸泼尼松、脊髓康可促进SCI后脊髓损伤区BDNF mRNA的表达,早期醋酸泼尼松效果较为明显,而干预后14 d脊髓康中剂量效果明显优于醋酸泼尼松。结论脊髓康可促进大鼠SCI后神经功能的恢复,提升SCI后脊髓内BDNF蛋白及mRNA的表达水平。
目的:探討脊髓康對脊髓損傷(SCI)後神經功能恢複以及腦源性神經營養因子(BDNF)蛋白和mRNA錶達的影響。方法144隻雌性Sprague-Dawley大鼠,體質量180~220 g,隨機抽取24隻作為假手術組,僅咬除T9~T11段椎闆、棘突。其餘採用改良Allen法成功建立SCI動物模型後,隨機分為5組,即模型組、醋痠潑尼鬆組及脊髓康高、中、低劑量組,每組24隻。脊髓康高、中、低劑量組分彆按生藥劑量50 g/(kg?d)、25 g/(kg?d)、12.5 g/(kg?d)灌胃;醋痠潑尼鬆組以醋痠潑尼鬆0.06 g/(kg?d)灌胃;假手術組與模型組均以同體積生理鹽水灌胃。分彆于術後24 h、3 d、7 d、14 d,進行Basso-Beattie-Bresnahan(BBB)評分、斜闆試驗,榦預後3 d、7 d、14 d處死大鼠取脊髓樣本,採用免疫組化、Western blotting、熒光定量PCR法檢測脊髓損傷區BDNF蛋白和mRNA的錶達情況。結果術後24 h假手術組大鼠BBB評分、斜闆試驗角度明顯高于其他各組(P<0.01)。術後3~14 d脊髓康中劑量組和醋痠潑尼鬆組BBB評分均高于模型組(P<0.05),且術後14 d脊髓康中劑量組BBB評分明顯高于其他各組(P<0.01)。免疫組化和Western blotting顯示,不同時間點醋痠潑尼鬆組和脊髓康中劑量組BDNF蛋白錶達水平均明顯高于模型組(P<0.01),二者具有等效性(P>0.05)。熒光定量PCR顯示,醋痠潑尼鬆、脊髓康可促進SCI後脊髓損傷區BDNF mRNA的錶達,早期醋痠潑尼鬆效果較為明顯,而榦預後14 d脊髓康中劑量效果明顯優于醋痠潑尼鬆。結論脊髓康可促進大鼠SCI後神經功能的恢複,提升SCI後脊髓內BDNF蛋白及mRNA的錶達水平。
목적:탐토척수강대척수손상(SCI)후신경공능회복이급뇌원성신경영양인자(BDNF)단백화mRNA표체적영향。방법144지자성Sprague-Dawley대서,체질량180~220 g,수궤추취24지작위가수술조,부교제T9~T11단추판、극돌。기여채용개량Allen법성공건립SCI동물모형후,수궤분위5조,즉모형조、작산발니송조급척수강고、중、저제량조,매조24지。척수강고、중、저제량조분별안생약제량50 g/(kg?d)、25 g/(kg?d)、12.5 g/(kg?d)관위;작산발니송조이작산발니송0.06 g/(kg?d)관위;가수술조여모형조균이동체적생리염수관위。분별우술후24 h、3 d、7 d、14 d,진행Basso-Beattie-Bresnahan(BBB)평분、사판시험,간예후3 d、7 d、14 d처사대서취척수양본,채용면역조화、Western blotting、형광정량PCR법검측척수손상구BDNF단백화mRNA적표체정황。결과술후24 h가수술조대서BBB평분、사판시험각도명현고우기타각조(P<0.01)。술후3~14 d척수강중제량조화작산발니송조BBB평분균고우모형조(P<0.05),차술후14 d척수강중제량조BBB평분명현고우기타각조(P<0.01)。면역조화화Western blotting현시,불동시간점작산발니송조화척수강중제량조BDNF단백표체수평균명현고우모형조(P<0.01),이자구유등효성(P>0.05)。형광정량PCR현시,작산발니송、척수강가촉진SCI후척수손상구BDNF mRNA적표체,조기작산발니송효과교위명현,이간예후14 d척수강중제량효과명현우우작산발니송。결론척수강가촉진대서SCI후신경공능적회복,제승SCI후척수내BDNF단백급mRNA적표체수평。
Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic fac-tor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away on-ly. The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg?d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg?d) Jisuikang respectively, which were given 20 ml/(kg?d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoper-ative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P<0.01). BBB scores were higher in both Groups C and E than in Group B 3 to 14 days after SCI (P<0.05), and was significantly higher in Group E than in the other groups 14 days after SCI (P<0.01). The results of immunohistochemistry and Western blotting showed that the protein expression of BDNF were significantly higher in Groups C and E than in Group B at different time points in the injured area after SCI (P<0.01), while there was no significant difference between Groups C and E (P>0.05). The results of RQ-PCR showed that prednisone and Jisuikang promot-ed the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.