神经酰胺诱导培养角质形成细胞分化的实验研究

目的：观察神经酰胺对无血清培养角质形成细胞分化的影响。方法：选择两种神经酰胺（C2和C6），分别添加于无血清培养的角质形成细胞液中，并在添加后0、3、6、12、24h分别回收总RNA，用逆转录PCR（RT－PCR）方法，检测角质形成细胞分化的特异性标记：TGase1、IVL、KRT、、KRT10等mRNA表达情况。结果：神经酰胺添加后，C2－神经酰胺对TGase1 mRNA、IVL mRNA表达在12小时均明显上升，在24小时分别升高达阴性对照的5.7倍和5.2倍；但C2－神经酰胺对KRT1 mRNA和KRT10 mRNA表达升高不明显。C6－神经酰胺对TGase1 mRNA、IVL mRNA表达大致与C2－神经酰胺相似，只是24小时表达比C2－神经酰胺降低，分别为阴性对照的3.3倍和3.5倍；但C6－神经酰胺对KRT1 mRNA和KRT10 mRNA表达显著高于C2－神经酰胺，24小时分别高达5.4倍和10倍。结论：神经酰胺能明显升高无血清培养的角质形成细胞对分化特异性标记蛋白mRNA的表达，诱导角质形成细胞的分化。

The impact of ceramide on differentiation of cultured keratinocytes

Objective:To investigate the impact of ceramide on the differentiation of keratinocytes cultured in serum-free medium.Methods: Two kinds of ceramide (C2 and C6) were selected and added into serum-free media with keratinocyte culture. In 0, 3,6,12 and 24 hour after adding ceramides, total RNA was abstracted to detect the expression of specific keratinocyte differentiation markers: TGasel, IVL, KRT1 KRT10, etc. by RT-PCR method.Results: The expression of TGasel mRNA and IVL mRNA was most significant in 12 hours in the media containing C2 ceramide, and the levels were 5.7 times and 5.2 times than that in the control media without C2 ceramide in 12 and 24 hours, respectively. The expression of KRT1 mRNA and KRT10 mRNA in the media containing C2 ceramide was also increased but did not reach a significant level. The expression of Tgasel mRNA and IVL mRNA in the media containing C6 ceramide was similar to the media containing C2 ceramide, and was 3.3 times and 3.5 times than that in control media in 24 hours. However, the expression of KRT1 mRNA and KRT10 mRNA was significanly higher in the media containing C6 ceramide than in the media containing C2 ceramide, with 10 in the former and 5.4 times in the later. Conclusion : Ceramide could significantly increase the expression of specific differentiation markers (protein mRNA) of keratinocytes cultured in serum-free media and could induce the differentiation of keratinocyte. [