PDBu对大鼠生长板软骨细胞增殖和分化的影响  

Effects of protein kinase C on proliferation,differentiation and SOX9 expression of rat growth plate chondrocytes in vitro

作　　者：季煜华[1] 曾耀英[1] 季秋虹[2] 

机构地区：[1]暨南大学组织移植与免疫研究中心,广东广州510632 [2]南通大学附属医院,江苏南通226001

出　　处：《基础医学与临床》2007年第9期961-965,共5页Basic and Clinical Medicine

基　　金：国家重点基础研究发展规划项目(G1999054303);国家自然科学基金(30600587);暨南大学引进人才启动基金(51205066)

摘　　要：目的探讨蛋白激酶C(PKC)对生长板软骨细胞增殖、细胞外基质合成和转录因子SOX9表达的影响。方法分离、培养大鼠肋生长板软骨细胞(RGC),分别采用倒置显微镜、同位素掺入、RT-PCR和Western blot方法检测1、10和100nmol/L的PKC激动剂PDBu对第一代无血清培养RGC的细胞形态、增殖、胶原和蛋白多糖的合成及COL2A1和AGGRECAN的转录及转录因子SOX9表达的影响。结果100nmol/LPDBu的处理,使无血清培养RGC的细胞形态接近于血清组(10%FBS);抑制增殖并促进胶原和蛋白多糖的合成,3H-TdR、3H-proline和35S-sulfate掺入量分别是无血清对照组的83%、152.6%和146.5%(P<0.05);促进COL2A1和AGGRECAN的转录和SOX9的表达。结论PKC的活化抑制了生长板软骨细胞增殖,促进其分化。Objective To investigate the effects of protein kinase C （PKC） on proliferation, extracelluar matrix synthesis and transcriptional factor SOX9 （ SRY-related high mobility group-box gene 9 ） expression of rat growth plate chondrocytes in vitro. Methods Rat costochondral growth plate chondrocytes （RGC） were isolated and cultured. The 1 s, serum free cultured passage RGCs were treated with 1, 10 and 100 nmol/L phorbol 12,13-dibutyrate （PDBu）, PKC agonist, cell morphology were observed with inverted microscopy, cell proliferation, COLLAGEN and GAG synthesis were detected by isotope incorporation COLLAGEN TYPE Ⅱ and AGGRECAN mRNA transcription and SOX9 expression were revealed by RT-PCR and Western blot. Results 100 nmol/L PDBu treatment made the cell morphology of serum free cultured RGC closed to serum group and inhibited proliferation but promoted COLLAGEN and AGGRECAN synthesis, ^3 H-TdR、 ^3 H-proline and ^35 S-sulfate incorporation of 100nmol/L PDBu group were as 83%, 52.6% and 146.5% as those of serum free control（ P 〈0.05 ）. COL2A1 ,AGGRECAN gene transcription and SOX 9 expression were also enhanced by 100 nmol/L PDBu treatment . There was no obvious effects of 1 nmol/L and 10 nmol/L PDBu on 1^st RGC. Conclusion Activated PKC inhibits growth plate chondrocyte proliferation and promotes differentiation.

关 键 词：蛋白激酶C 生长板 软骨细胞 增殖 分化 SOX9 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]