低强度脉冲超声经PI3K/Akt通路调控兔膝骨性关节炎软骨细胞凋亡的作用机制研究  

The action mechanism of low intensity pulsed ultrasound regulating chondrocytes apoptosis in rabbits with osteoarthritis via activating PI3K/Akt pathway

作　　者：任莎莎[1] 李雪萍[1] 林强[1] 程凯[1] 夏鹏[1] 杨怀春[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：《中国康复》2015年第2期83-87,共5页Chinese Journal of Rehabilitation

基　　金：国家自然科学基金(81272151)

摘　　要：目的:探讨低强度脉冲超声(LIPUS)对兔膝骨性关节炎(OA)软骨细胞凋亡的影响及有关作用机制。方法:共选取30只新西兰大白兔,随机分为正常对照组(A组)、OA模型组(B组)、OA+LIPUS组(C组)、OA+LY294002(PI3K/Akt抑制剂)组(D组)、OA+LY294002+LIPUS组(E组),每组各6只。采用右后膝前交叉韧带切断术(ACLT)造模。于造模后第6周时采用空气栓塞法处死实验兔,切取软骨组织进行组织学观察,并进行Mankin评分;提取软骨细胞进行体外培养并采用免疫组化染色法鉴定;应用western blot检测各组软骨细胞中Ⅱ型胶原(COL2)、MMP-13、Akt、pAkt、P53、Bcl-2蛋白表达情况。结果:B组COL2、pAkt、Bcl-2蛋白含量较A组降低,MMP-13、P53蛋白含量较A组增高(P<0.05)。与B组相比,C组COL2、pAkt、Bcl-2蛋白含量增高,MMP-13、P53蛋白含量下降(P<0.05);D组COL2、Bcl-2、pAkt蛋白含量下降,MMP-13、P53蛋白含量升高(P<0.05),E组COL2、MMP-13、P53、Bcl-2、pAkt蛋白含量无明显变化,但与C组、D组相比差异显著(P<0.05)。各组Akt蛋白表达无明显差异。结论:LIPUS能通过PI3K/Akt通路降低兔膝骨性关节炎软骨细胞凋亡率,促进抗凋亡基因Bcl-2表达,下调促凋亡基因P53水平,促进关节软骨损伤修复。Objective：To investigate the effects of low intensity pulsed ultrasound（LIPUS）on the apoptosis of chondrocytes in rabbits with osteoarthritis（OA）and explore the underlying mechanism.Methods：Thirty healthy New Zealand white rabbits were selected for this study.All the cultured cells were randomly divided into five groups：a normal control group（A group）,an OA model group（B group）,an OA model plus LIPUS group（C group）,an OA model plus PI3K/Akt inhibitor group（D group）,and an OA model plus PI3K/Akt inhibitor and LIPUS group（E group）.The OA model was made by knee anterior cruciate ligament transection.At the 6th week after modeling,the rabbits were sacrificed by air embolism method and pathologic changes were assessed on articular surface of femoral condyles.Meanwhile chondrocytes were isolated and cultured in vitro,and cultured cells were identified by immuohistochemistry.The expression of collagen protein typeⅡ,MMP-13,Akt,pAkt,P53,and Bcl-2was detected by Western blotting.Results：As compared with the A group,the expression of collagen type Ⅱ,pAkt and Bcl-2was significantly reduced in the B group,but that of MMP-13 and P53was significantly increased in the B group（P〈0.05）.As compared with the B group,the expression of collagen type Ⅱ,pAkt and Bcl-2was significantly increased,and that of MMP-13 and P53was significantly reduced in the C group（P〈0.05）;the expression of collagen typeⅡ,pAkt and Bcl-2was significantly reduced（P〈0.05）,and that of MMP-13 and P53was significantly increased in the D group（P〈0.05）.Although the expression of collagen typeⅡ,MMP-13,pAkt,P53,and Bcl-2showed no significant difference between the E group and the B group,the expression of collagen type Ⅱ, MMP-13,pAkt,P53 and Bcl-2showed significant difference between C group and D group（P〈0.05）.There was no significant difference in the expression of Akt among all groups.Conclusions：The LIPUS can obviously decrease the apoptosis rate of chondrocytes in the knee joints of O

关 键 词：低强度脉冲超声 软骨细胞 PI3K/AKT通路 凋亡 

分 类 号：R445.1[医药卫生—影像医学与核医学] R684.3[医药卫生—诊断学]