Akt与ERK1/2在人骨关节炎软骨细胞中的表达  

Expression of Akt and ERK1/2 in human osteoarthritic chondrocytes

作　　者：程亮[1] 赵洪海[2] 曾国庆[2] 张同恩[2] 王少杰[2] 石磊[2] 王承云[2] 夏春[1,2] 

机构地区：[1]福建医科大学协和临床医学院,福建福州350000 [2]厦门大学附属中山医院骨关节科,福建厦门361000

出　　处：《中国病理生理杂志》2012年第5期889-894,共6页Chinese Journal of Pathophysiology

基　　金：福建省自然科学基金资助项目(No.2010D007);福建省医学创新课题资助(No.2011-CXB-36)

摘　　要：目的:观察蛋白激酶B(Akt)与细胞外信号调节激酶1/2(ERK1/2)在正常和骨关节炎(OA)软骨细胞中的表达,探讨Akt与ERK1/2在OA病程中的意义。方法:手术中取5例正常和18例OA人膝关节软骨组织,包埋制备切片,免疫组织化学技术观察p-Akt及p-ERK1/2在正常和OA软骨组织中的表达;培养人软骨细胞,甲苯胺蓝染色、免疫组化鉴定并观察聚集蛋白聚糖及Ⅱ型胶原在正常和OA软骨细胞中的表达;Western blot-ting技术检测Akt、p-Akt、ERK1/2、p-ERK1/2、磷酸化70 kD核糖体蛋白S6激酶(p-p70S6K)及增殖细胞核抗原(PCNA)蛋白在正常和OA软骨细胞中表达水平;实时荧光定量PCR技术检测聚集蛋白聚糖及Ⅱ型胶原在正常和OA软骨细胞中mRNA表达水平。结果:与正常软骨细胞比较,OA软骨细胞内p-Akt和p-p70S6K蛋白表达明显降低(P<0.05),且聚集蛋白聚糖和Ⅱ型胶原mRNA和蛋白在OA软骨细胞中的表达水平降低(P<0.05),而p-ERK1/2和PCNA蛋白表达明显提高(P<0.05)。结论:Akt可能通过p-p70S6K来调控OA软骨细胞外基质聚集蛋白聚糖及II型胶原的合成,ERK1/2可能通过PCNA来调控OA软骨细胞增殖;Akt与ERK1/2可能参与了OA的病理过程。AIM： To investigate the expression of protein kinase B （Akt） and extracellular signal -regulated kinase 1/2 （ERK1/2） in normal and osteoarthritic chondrocytes. METHODS： The samples of knee cartilage were ob- tained from the normal donors （n ---5 ） and the patients （n = 18） undergoing total knee arthroplasty with the diagnosis of os- teoarthritis （OA）. The expression of p -Akt and p - ERK1/2 in the normal and osteoarthritic cartilage tissues was detec- ted by the method of immunohistochemistry. The chondrocytes were isolated and identified by toluidine blue staining and immunohistochemical method. The expression levels of Akt, p - Akt, ERK1/2, p - ERK1/2, phosphorylated 70 - kD ribo- somal protein S6 kinase（ p - p70S6K） and proliferating cell nuclear antigen（PCNA） were tested in normal and osteoarthrit- ic chondrocytes by Western blotting. Real - time fluorescence quantitative PCR was used to measured the expression levels of aggrecan and type II collagen gene in normal and osteoarthritic chondrocytes. RESULTS： The expression of p - Akt in normal cartilage was higher than that in OA cartilage. The expression of p - ERK1/2 in OA cartilage was higher than that in normal cartilage. Compared with the normal chondrocytes, the expression of p -Akt and p -p70S6K, and the mRNA levels of aggrecan and type II collagen were increased （P 〈0. 05）, and the expression of p - ERK1/2 and PCNA was de- creased in OA chondrocytes （P 〈 0. 05）. CONCLUSION： Akt might regulate aggrecan and type II collagen synthesis via p- p70S6K, and ERK1/2 might regulate OA chondrocyte proliferation through PCNA. Both Akt and ERK1/2 play impor- tant roles in the pathogenesis of OA.

关 键 词：骨性关节炎 软骨细胞 蛋白激酶B 细胞外信号调节MAP激酶类 蛋白合成 细胞增殖 

分 类 号：R363.1[医药卫生—病理学]