摘要
目的:探讨不同浓度的表皮生长因子(EGF)对传代培养的大鼠肋生长板软骨细胞(RGC)增殖和胶原合成的影响。方法:分离、培养RGC,分别用Westernblot和阿尔新蓝(Alcineblue)染色检测各代RGC中Ⅱ型胶原和蛋白多糖的表达;用[3H]-TdR和[3H]-proline掺入分别检测1、10和100μg/LEGF对第1、3和5代RGC增殖和胶原合成的影响。结果:原代培养的RGC表达Ⅱ型胶原和蛋白多糖,从第4代起Ⅱ型胶原和蛋白多糖的表达迅速降低,RGC发生了去分化。[3H]-TdR掺入表明,EGF促进第1代RGC的增殖(P<0.01),3个浓度的作用依次为:1μg/L>10μg/L>100μg/L,差异显著(P<0.01);3个浓度的EGF对第3代RGC保持了相近的促增殖作用(P<0.01);对第5代RGC均无促增殖作用(P>0.05)。与促增殖作用不同,不同浓度EGF促不同传代的RGC的[3H]-proline掺入率比对照组均高20%左右(P<0.01)。结论:EGF具有促进培养RGC增殖和胶原合成的作用,连续传代引起的去分化降低了RGC对EGF促增殖作用的反应,但对EGF的促胶原合成作用没有产生影响。
AIM： To evaluate the effects of EGF on proliferation and collagen synthesis of serial passaged rat costochondral growth plate ehondrocytes （ RGCs）.METHODS： RGCs were isolated and cultured, collagen type Ⅱ and proteoglyean expression in serial passaged RGCs were detected by Western blotting and Alcine blue staining. The effects of 1μg/L, 10μg/L, 100 μg/ L EGF on passage 1, 3 and 5 RGC proliferation and collagen synthesis were measured by [^3H] - TdR and [^3H] - prohne incorporation.RESULTS： Primary cultured RGCs expressed type Ⅱ collagen and proteoglycan. Since passage 4, the expression of type Ⅱ collagen and proteoglycan decreased abruptly and RGC dedifferentiation was observed. [^3H] - TdR incorporation demonstrated that all the three concentrations of EGF enhanced passage 1 RGC proliferation （ P 〈 0.01 ）. Their proliferative efficieneies were： 1μg/ L〉 10μg/L〉 100μg/L. EGF kept its proliferative stimulation on passage 3 RGCs （ P 〈 0.01）, but there was no difference among the three treated groups （ P 〉 0.05）. EGF lost its proliferative effects on the passage 5 RGCs（ P 〉 0.05）. Unlike the proliferative effects of EGF, there was no difference among testing groups in passage 1, 3 and 5 RGCs in collagen synthesis enhancement （ P 〉 0.05）. [^3H] - proline incorporation in testing groups was 20% higher than that in control. CONCLUSION： The present study suggests EGF is able to enhance RC, Cs proliferation and collagen synthesis. Dedifferentiation caused by serial passage decreases proliferative effect of EGF on RGCs, but has no effect on collagen synthesis enhancement.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第3期592-596,共5页
Chinese Journal of Pathophysiology
基金
973国家重点基础研究发展规划项目(No.G1999054303)
广东省"十五"重大科技专项(No.A302020204)
关键词
软骨细胞
生长面
表皮生长因子
胶原
Chondrocytes
Growth plate
Epidermal growth factor
Collagen
