低强度脉冲超声对兔膝骨性关节炎软骨细胞整合素-局部粘着斑激酶-促分裂原活化蛋白激酶力化学转导通路相关蛋白表达的影响  

The effects of low intensity pulsed ultrasound on expression of mechanochemical transduction pathway-related protein of integrin-focal adhesion kinase- mitogen- activated protein kinases in the chondrocytes of rabbits with knee osteoarthritis

作　　者：夏鹏[1] 李雪萍[1] 林强[1] 程凯[1] 沈士浩[1] 任莎莎[1] 高明霞[1] 

机构地区：[1]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

出　　处：《中华物理医学与康复杂志》2014年第3期165-170,共6页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金（81272151）

摘　　要：目的观察低强度脉冲超声（LIPUS）对兔膝骨性关节炎（OA）软骨细胞中整合素。局部粘着斑激酶（FAK）-促分裂原活化蛋白激酶（MAPK）力化学信号转导通路相关蛋白表达的影响。方法共选取18只新西兰大白兔，其中12只进行膝OA造模，其余6只作为正常对照。于制模后第4周处死实验兔，提取软骨细胞进行体外培养并鉴定。将所培养细胞分为正常对照组、OA模型组及OA超声组。待细胞传至第2代时，正常对照组及OA模型组细胞不给予任何处理，OA超声组细胞则给予LIPUS辐射。于LIPUS持续作用1周后收集各组细胞，应用Westernblot技术检测各组细胞中Ⅱ型胶原、蛋白多糖、基质金属蛋白酶（MMPs）．13、整合素B1、FAK、MAPK（如p38、ERKl／2、JNK）蛋白表达以及磷酸化蛋白表达情况。结果①OA模型组及OA超声组Ⅱ型胶原、蛋白多糖含量均较正常对照组明显降低（P〈0．05），并以OA模型组下降幅度较显著，与OA超声组间差异具有统计学意义（P〈0．05）。②OA模型组及OA超声组MMP一13含量均较正常对照组明显增高（P〈0．05），并以OA模型组增高幅度较显著，与OA超声组间差异具有统计学意义（P〈0．05）。③OA模型组及OA超声组整合素pl、磷酸化FAK含量均较正常对照组明显增高，且以OA超声组增高幅度较显著，与OA模型组问差异具有统计学意义（P〈0．05）。④OA模型组磷酸化p38、ERKl／2及JNK含量均较正常对照组明显增加（P〈0．05），OA超声组上述指标则较OA模型组显著降低（P〈0．05）。结论LIPUS体外辐射可抑制OA软骨细胞Ⅱ型胶原、蛋白多糖降解及MMP-13表达，同时还能促进整合素Bl表达以及FAK磷酸化，抑制p38、ERKl／2、JNK磷酸化，提示LIPUS辐射可能通过启动整合素一FAK—MAPK力学信号转导通路诱导软骨细胞外基质发生改变。Objective To observe any effect of low intensity pulsed ultrasound （LIPUS） on the expres- sion of integrin-focal adhesion kinase （FAK）- mitogen-activated protein kinase （MAPK） mechanochemical trans- duetion pathway-related proteins in the ehondrocytes of rabbits with knee osteoarthritis （OA）. Methods Of the 18 New Zealand white rabbits selected for the study, twelve received knee anterior cruciate ligament transection to model OA. The remaining 6 rabbits served as normal controls. At the 4th week after modeling the rabbits were sac- rificed and chondrocytes were isolated and cultured in vitro. All the cultured cells were randomly divided into three groups： a normal control group （NC） , an OA model group （OA） and an OA model plus LIPUS group （ OA ＋ LIPUS）. When the cells had been cultured to the 2nd passage, the NC group and OA group cells had received no treatment. The OA ＋ LIPUS group cells were exposed to 40 mW/cm2 of LIPUS for 20 min, once a day for 6 days. The expression of collagen protein type II, aggrecan, MMP-13, integrin 131 p-FAK and p-p38, p-ERK1/2 and p-JNK Mapks were detected by Western blotting. Results Compared with the NC group, the expression of colla- gen type II and aggrecan was significantly lower in the OA and OA ＋ LIPUS groups, with more significantly lower expression of collagen type II and aggrecan in the OA group than in the OA ＋ LIPUS group. Compared with the NC group, the expression of MMP-13 was significantly higher in the OA and OA ＋ LIPUS groups, with a significantly larger increase in the OA group. Compared with the NC group, the expression of integrin 131 and p-FAK was also significantly higher in the OA and OA ＋ LIPUS groups, with a significantly larger increase in the OA ＋ LIPUS group. Compared with the NC group, the expression of p-p38, p-ERK1/2 and p-JNK was also significantly higher in the OA group, but compared with the OA group, the expression of those kinases was, on average, significantly lower in the OA ＋ LIPUS group. Conclusi

关 键 词：低强度脉冲超声 软骨细胞 整合素 局部粘着斑激酶 促分裂原活化蛋白激酶 

分 类 号：R684.3[医药卫生—骨科学]