角质形成细胞条件培养液对成纤维细胞增殖与胶原分泌影响的研究

EFFECTS OF CONDITIONED MEDIUM DERIVED FROM KERATINOCYTES ON FIBROBLASTS

目的探讨正常的角质形成细胞对成纤维细胞生物学行为的影响。方法组织块法培养成纤维细胞,分别加入12.5%、25%与50%浓度的角质形成细胞条件培养液为实验组,加入不含血清DMEM为对照组,采用MTT、羟脯氨酸比色法测定成纤维细胞增殖、胶原合成;以流式细胞仪测定50%浓度角质形成细胞条件培养液组及对照组成纤维细胞生长周期。结果细胞增殖测定,各实验组吸光度(A)值与对照组比较,差异均有统计学意义(P<0.01);随浓度增高,A值增加,25%及50%浓度组与12.5%浓度组比较,差异有统计学意义(P<0.01);25%及50%浓度组间比较,差异无统计学意义(P>0.01)。胶原合成测定中,各实验组A值与对照组比较,差异无统计学意义(P>0.01);各实验组组间比较,差异无统计学意义(P>0.01)。细胞周期测定见,50%浓度组可明显促进成纤维细胞通过G1/S及S/G2限制点,S期及G2/M期细胞与对照组相比明显增多,G0/G1期细胞与对照组相比明显减少,差异均有统计学意义(P<0.01)。结论正常角质形成细胞的上清液可促进成纤维细胞的增殖,但对其胶原分泌影响不明显。

Objective To observe the effects of keratinocytes on proliferation and collagen secretion of fibroblasts. Methods The conditioned medium, collected from cultured keratinocytes, was added to the cultured fibroblasts as the tested groups （12. 5%, 25% and 50% groups） and DMEM as control group. The MTT, bydroxyproline coloricmetric method and flow cytometer were employed to measure the fibroblast proliferation, the collagen secretion and the change of the cell cycle. Results In fibroblast proliferation, the absorbency（A） value of tested groups was significantly different from that of the control group （P〈0. 01）. A value increased as increasing concentration, there was statistically significant difference between the concentrations of 25%, 50% and the concentration of 12. 5%（P〈0.01）, but no statistically significant difference between the concentrations of 25% and 50% （P〈0.01）. In collagen secretion, there was no statistically significant difference between the tested groups and the control group（P〈0. 01）, and between the tested groups（P〈0. 01）. In cell cycle, 50% of conditioned medium could make the fibroblast pass the limit of G1/S and S/G2 period, the cell rates of S,G2-M period increased. Conclusion The conditioned medium from keratinocytes can increase fibroblasts proliferation, have little effect on general collagen secretion.