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目的:探讨龙血竭不同有效部位对体外培养的胎鼠皮肤成纤维细胞生长及分泌的影响。方法:孕龄12.5~14.5d昆明小鼠胚胎,无菌条件下将躯干部皮肤制成匀浆,接种于25cm2塑料培养瓶,体积分数10%FBS的高糖DMEM培养基培养传代,P3代细胞以8×106个/L接种于96孔板,细胞饥饿24h后进行实验。采用石油醚、氯仿、乙酸乙酯、正丁醇、乙醇等极性递增溶剂依次萃取龙血竭中的各种有效部位,均选用0.125g/L、0.25g/L、0.5g/L、1g/L、2g/L、4g/L五个浓度梯度作用于96孔板中的细胞(DMSO助溶),24h后MTT法检测不同条件下细胞的生长曲线,免疫组化检测细胞分泌Ln、Ⅰ型胶原的变化。10%FBS的高糖DMEM培养条件为空白对照,1%DMSO的高糖DMEM培养条件为实验对照。结果:龙血竭乙酸乙酯有效部位在低浓度对胎鼠成纤维细胞的生长有轻度抑制作用,随浓度增加促生长作用增强,1g/L龙血竭乙酸乙酯有效部位对胎鼠皮肤成纤维细胞的促生长作用最佳,并促进Ln、Ⅰ型胶原的分泌。龙血竭其它有效部位对胎鼠成纤维细胞均无刺激生长的作用。结论:龙血竭乙酸乙酯有效部位能促进胎鼠皮肤成纤维细胞的生长和分泌。
Objective: To investigate the effects of different effective parts of DRG on the growth and secretion of fetal rat skin fibroblasts cultured in vitro. METHODS: The Kunming mouse embryos at the gestational age of 12.5-14.5 days were homogenized under aseptic conditions and inoculated into 25cm2 plastic culture flasks and cultured in high glucose DMEM medium with volume fraction of 10% FBS. P3 generation cells To 8 × 106 / L inoculated in 96-well plates, the cells were starved for 24h after the experiment. The effective fractions of Rhodiola sachalinensis were extracted by petroleum ether, chloroform, ethyl acetate, n-butanol, ethanol and other polar increasing solvents in succession. All the selected fractions were 0.125g / L, 0.25g / L, 0.5g / The cells were cultured in DMSO (5%) / L, 2g / L and 4g / L concentration gradient. The growth curves of cells in different conditions were detected by MTT assay 24h later. Collagen changes. 10% FBS high glucose DMEM culture conditions for the blank control, 1% DMSO high glucose DMEM culture conditions for the experimental control. Results: The effective area of ethyl acetate of Dracaena Sinensis had a slight inhibitory effect on the growth of fetal rat fibroblasts at low concentration, and the growth promoting effect was enhanced with the increase of concentration. The effect of ethyl acetate at 1g / L on fetal mouse skin Fibroblasts promote the best growth and promote Ln, type Ⅰ collagen secretion. Dragon’s blood other effective parts of fetal rat fibroblasts no stimulation of growth. Conclusion: The effective fraction of ethyl acetate in dragon’s blood can promote the growth and secretion of fetal rat fibroblasts.