| 高明霞,程 凯,林 强,王大新,于俊龙,林爱翠,安恒远,陈安亮,李雪萍.低强度脉冲超声波对早中期兔膝骨性关节炎软骨细胞外基质及MAPKs信号通路的影响[J].中国康复医学杂志,2013,28(7):593~599 |
| 低强度脉冲超声波对早中期兔膝骨性关节炎软骨细胞外基质及MAPKs信号通路的影响 点此下载全文 |
| 高明霞 程 凯 林 强 王大新 于俊龙 林爱翠 安恒远 陈安亮 李雪萍 |
南京医科大学附属南京医院(南京市第一医院)
康复医学科，南京，210006 |
| 基金项目：国家自然科学基金资助项目（81272151） |
| DOI： |
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| 摘要： |
| 摘要
目的：研究低强度脉冲超声波（LIPUS）对早中期兔膝骨性关节炎（OA）软骨细胞外基质（ECM）、丝裂原活化蛋白激酶（MAPKs）信号通路的影响，探讨LIPUS对关节软骨损伤修复和延缓退变的作用机制。
方法：36只健康新西兰兔随机分成6组，早期对照组（EC组）、早期OA组（EO组）、早期治疗组（ET组）、中期对照组（MC组）、中期OA组（MO组）和中期治疗组（MT组），6只/组。EO、ET、MO及MT组均接受左后肢前交叉韧带切断术（ACLT），对照组仅接受左侧膝关节囊切开术。ET组和MT组分别于术后第3天和第5周起接受LIPUS治疗。超声频率3MHz，强度40mW/cm2，作用时间20min，1次/d，6d/周，持续6周。EO与MO组的LIPUS方案与治疗组相同，但无超声输出。LIDUS 6周后，采用甲苯胺蓝染色进行关节软骨的组织学观察，并进行Mankin评分。采用免疫印迹法检测蛋白多糖、Ⅱ型胶原、磷酸化的细胞外信号调节激酶1/2（p-ERK1/2）及p38（p-p38）的变化。
结果：①组织学观察及Mankin评分：EO组关节软骨表面不规则、甲苯胺蓝染色变浅、软骨细胞减少，与EC组相比，Mankin评分显著升高（P<0.01）；MO组关节软骨损伤明显，Mankin评分较MC组显著升高（P<0.01）。与EO组相比，ET组病理学改变程度轻，Mankin评分显著降低（P<0.01）；但MT组Mankin评分较MO组无显著降低（P>0.05）。②Ⅱ型胶原、蛋白多糖检测：与EC组比较，EO组和ET组均下降，但EO组较ET组明显下降（P<0.05）；与MC组相比，MO和MT组均显著降低（P<0.05），MO组与MT组间无显著差异（P>0.05）。③p-ERK1/2、p-p38检测：与EC组相比，EO组表达量显著升高（P<0.05），但与EO组相比，ET组显著降低（P<0.05）；与MC组相比，MO组和MT组表达显著升高（P<0.05），MT组与MO组间无明显差异（P>0.05）。
结论：LIPUS可以减轻关节软骨ECM的损伤程度，其作用与LIPUS治疗后关节软骨中p38、ERK1/2表达下调有关，这种作用与OA的病变阶段密切相关，即在OA早期进行LIPUS作用更明显。 |
| 关键词：低强度脉冲式超声波 骨性关节炎模型 细胞外基质 丝裂原活化蛋白激酶 丝裂原活化蛋白激酶信号通道 |
| Effect of low-intensity pulsed ultrasound on extracellular matrix and MAPKs signaling pathways in early and medium term of rabbit knee osteoarthritis model Download Fulltext |
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| Dept. of Rehabilitation, Nanjing First Hospital, Nanjing Medical University，210006 |
| Fund Project: |
| Abstract: |
| Abstract
Objective：To establish rabbit osteoarthritis（OA） model by anterior cruciate ligament transaction and to observe the articular cartilage repair effects of low-intensity pulsed ultrasound（LIPUS） and to explore it's mechanism.
Method：Thirty-six healthy New Zealand rabbits were randomized into early control（EC） group, early osteoarthritis（IA） group, early treatment（ET） group, medium-term control（MC） group, medium-term osteoarthritis（MO） group and medium-term treatment（MT） group. Animals in EC group and MC group underwent articular capsulotomy surgery, and those in other four groups underwent anterior cruciate ligament transection（ACLT）. In EO group and ET group, rabbits received LIPUS at 3d after surgery, while in MO group and MT group received LIPUS at the beginning of the 5th week after surgery. LIPUS treatment was applied with 3MHz, 40mW/cm2, for 20min, 1times/d, 6d/week, total 6 weeks. However, in EO group and MO group had no LIPUS output. Six weeks after LIPUS, the rabbits were sacrificed and pathologic changes of articular surface of femoral condyle were assessed. Also, expressions of typeⅡcollagen, proteoglycan, phophorylated extracellular signal-regulated kinase1/2（P-ERK1/2） and p38 of articular cartilage were measured with Western blot.
Result：①Histopathological observations and Mankin scores: In comparison with EC group, in EO group, the surfaces of articular cartilage were irregular and not smooth, toluidine blue staining became light, the amounts of cartilage cells were less, Mankin scores increased significantly(P<0.01); in MO group, damages of cartilage were obvious, Mankin scores were higher than that of MC group(P<0.01). In comparison with EO group, pathological changes in ET group were lighter, Mankin scores reduced significantly(P<0.01); but compared with MO group, Mankin scores of MT group had no significant decrease(P>0.05).②The expressions of typeⅡcollagen, proteoglycan in Western blot analysis: In comparison with EC group, the expressions of type Ⅱcollagen and proteoglycan in EO and ET groups both decreased, but that in ET group were significantly higher than that in EO group (P<0.05); In comparison with MC group, the expressions of both in MO and MT groups decreased (P<0.05), and no difference was found between MO and MT group (P>0.05).③The expressions of p-ERK1/2 and p-p38 with Western blot analysis: In comparison with EC group, the expressions of p-ERK1/2 and p-p38 in EO group increased significantly(P<0.05), and no difference in EC group (P>0.05). In comparison with EO group, the expressions of p-ERK1/2 and p-p38 in ET group decreased significantly(P<0.05). While in comparisons with MC group, the expressions of p-ERK1/2 and p-p38 in MO and MT groups increased significantly(P<0.05).
Conclusion：Low-intensity pulsed ultrasound intervention is beneficial to articular cartilage repair by abating damages of extracellular matrix in early term of osteoarthritis， and extracellular signal-regulated kinase1/2 and p38 signaling pathway was involved. |
| Keywords:low-intensity pulsed ultrasound osteoarthritis model extracellular matrix mitogen-activated protein kinases mitogen-activated protein signaling pathways |
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