蛋白激酶b与蛋白激酶c在人骨性关节炎软骨组织中的表达的分析研究.pdf

蛋白激酶B与蛋白激酶C在人骨性关节炎软骨组织中的表达研究摘要目的:观察并研究PKB/Al(t和PKC在人膝骨性关节炎(osteoarthritis,OA) 关节软骨中的表达及其与软骨细胞凋亡的相关性,探讨PKB/Akt和PKC信号分子在OA发病中的作用,为从分子水平治疗OA提供新靶点。方法:选取38例因膝骨性关节炎行膝关节置换的患者为实验组,以ll例因意外受伤行髁上截肢手术的患者为对照组,两组均从股骨内髁负重区取软骨组织。,;免疫荧光法和免疫组化法观察PKB/Akt和PKC在软骨细组织的分布:TUNEL法检测软骨细胞的凋亡状况;免疫组化法和WestemBlotting法观察关节软骨PKB/Akt、 p-Akt和PKC的表达。 。采用随机两样本t检验、配对两样本t检验、Pearson相关性检验进行统计分析。结果:①组织形态学观察:正常关节软骨组织厚度正常,浅表层表面光滑、平整,软骨细胞排列整齐,潮线完整,无细胞外基质丢失;OA关节软骨组织厚度变薄,浅表层粗糙、破溃或消失,软骨细胞排列紊乱,潮线断裂,细胞外基质严重丢失。②软骨细胞凋亡计数结果:OA软骨组织中软骨细胞的凋亡率高于正常(t--,P<);无论OA还是正常软骨组织过渡层的凋亡率均高于放射层(,t=,p<)。③软骨组织中PKB/Akt及 PKCa的表达:PKB/AI(t在正常软骨组织中的表达高于在OA(, P<),而磷酸化的PKB/Akt()却在OA软骨组织中表达较高;PKCa 在正常软骨组织中的表达低于OA(r--,P<)。④软骨组织中PKB/Akt 及PKCa的分布:无论在正常还是在OA关节软骨组织中,PKB/Akt和PKCa 主要分布在过渡层和放射层,过渡层PKB/Akt和PKCa的表达均高于放射层, 其差异具有显著性,统计学数据依次为:PKB/Akt t-"-,P<,PKCa t=,P<;PKB/Akt t=,P<,,P<。(9PKB/Al(t 和PKCa主要分布在软骨细胞的细胞浆内。结论:软骨细胞PKB/Akt表达减少,PKCa表达增加是OA软骨细胞凋亡增强的分子基础。PKB/Akt和PKCa具有临床预防和治疗OA的新靶点的潜能。关键词:骨性关节炎;软骨组织;软骨细胞;凋亡;PKB/Akt:PKCa 2 The study on theexpression ofprotein kinase Band protein kinase Cinhuman teoarthritis cartilal臣,Drotetn nase in man osteoart riUs cartila翌[e Abstract obiective Withdetecting theexpression and relationship ofPKB/Akt and PKC inkneeosteoarthritis(OA),We investigating theroleofPKB/Akt and PKC in thepathogenesis ofOA,in order toprovide some evidence forclinicaltherapy at themolecular level. Methods 38samples ofOA human cartilage were collected from totalknee arthroplasty as experimental samples of normal human cartilage were collected from patients were amputated because ofunintentional injuries as samples were selected fromthecondyle normal HE and safranin-o staining were used toobserve thehistological paRems of articularcartilage;the distribution ofPKB/Akt and PKC inthe chondrocyte were observed under Laser scanning confocai metheod were