中药白芨和地榆对角质形成细胞游走的不同影响

Different Influence of Rhizoma Bletillae and Garden Burnet on Keratinocyte Migration

目的 探讨角质形成细胞游走在表皮病理修复中的作用及白芨和地榆对角质形成细胞游走的不同影响。方法 采用无血清表皮器官培养法 ,观察角质形成细胞正常游走时的形态变化和游走长度 ,以及含不同浓度白芨或地榆的培养剂对角质形成细胞游走的影响。结果 角质形成细胞从切缘基底层开始游走 ,形成皮包。核深染 ,未见核分裂相 ,胞浆染色较红 ,未见角质层。皮包起始部呈多层细胞 ,远端渐呈单层细胞。游走长度与培养时间呈直线正相关关系 ,与直线拟合 (P >0 .0 5 ) ,培养 2 4、4 8、72h游走长度皆有显著增加 ,组间差异非常显著 (P <0 .0 1 )。不同浓度含白芨上清液成分无血清培养基培养表皮植片 ,2 4、4 8h后表皮游走长于正常对照组。白芨浓度为 2× 1 0 -1mg/ml时差异无显著性 (P >0 .0 5 ) ,浓度为 2× 1 0 -2 mg/ml及 2× 1 0 -3 mg/ml时 ,促游走作用明显优于对照组 ,差异非常显著 (P <0 .0 1 )。浓度稀释到 2× 1 0 -4mg/ml时 ,游走接近于正常对照组。白芨培养 4 8h游走皆显著长于培养 2 4h。不同浓度含地榆上清液成分无血清培养基培养表皮植片 ,2 4、4 8h后部分表皮游走低于正常对照组。地榆浓度为 2× 1 0 -1 mg/ml时游走长度非常显著地低于正常对照组 (P <0 .0 1 )。浓度稀释为 2× 1 0 -2 mg/ml。

Objective To study the effect of keratinocyte migration on epidermis healing and the different influence of Rhizoma Bletillae(RB) and Garden Burnet(GB) to keratinocyte migration.Methods Non blood serum epidermis organ culture was adopted.The morphologic change and migration distance of normal keratinocyte and the different influence of RB and GB in different concentration to keratinocyte migration were observed.Results Keratinocyte started to migrate from the cut edge of basal layer and formed epiboly which showed deep stained nucleus without karyokinesis and red stained cytoplasm without cuticular layer.It also showed poly layer of keratinocytes at the close end of epiboly and mono layer at the far end.There was a positive linear correlation between the migration distance and culture time ( P >0.05).There was a significant increase of migration distance at 24,48 and 72 h cultivation ( P <0.01).Epidermal explants cultivated in culture medium without blood serum and with different percent of RB supernatant gradients for 24 h or 48 h showed longer migration distance than controls.Explants in medium of RB in 2×10 -1 mg/ml did not migrate much longer than controls( P >0.05);but explants in medium of RB in 2×10 -2 mg/ml or 2×10 -3 mg/ml migrated significantly longer than controls( P <0.01);but explants in 2×10 -4 mg/ml,close to controls.Explants in different concentrations of RB after 48 h culture migrated significant longer than 24 h culture.Epidermal explants cultivated in culture medium without blood serum and with different percent of GB supernatant gradients for 24 h or 48 h showed shorter migration distance than controls.Explants in medium of GB in 2×10 -1 mg/ml migrated very significant shorter than controls( P <0.01);in 2×10 -2 mg/ml,2×10 -3 mg/ml or 2×10 -4 mg/ml,all close to controls( P >0.01).Conclusion There is a series of biologic and morphologic changes in cell form,cytoplasma and nucleus that fit for migration as keratinocytes transfer to migration cells.The migration distance of keratinocytes showed positive linear correlation with culture time.RB significantly promotes migration that is possibly related to its effect in treating ulceration and trauma.GB significantly inhibits migration that means its effect in trauma treatment is not related to keratinocyte migration.