CAJ | 학술논문

目的探讨女贞子乙醇提取物及其单体酪醇对人表皮黑素细胞黏附和迁移的调节作用.方法体外分离培养人表皮黑素细胞,XTT法测定细胞增殖情况,用经FN包被的培养板测定细胞黏附率,Transwell微孔膜法观测细胞迁移情况,激光共聚焦显微镜观察经中药处理的细胞内肌动蛋白细胞骨架的结构分布,半定量分析细胞内荧光强度.结果 0.0375～0.6 mg/ml女贞子乙醇提取物均可促进黑素细胞在FN上的黏附(P＜ 0.05或P＜ 0.01);0.5～2 mmol/L酪醇可促进黑素细胞黏附(P＜0.05或P＜0.01).0.15 mg/ml女贞子乙醇提取物及2 mmol/L酪醇无明显细胞毒性,在24 h时不明显促进细胞增殖,选择此浓度为工作浓度,女贞子乙醇提取物及酪醇穿过微孔滤膜的黑素细胞数分别为43.7和51.0个,显著高于对照组的20.3个(P＜ 0.01);两种药物工作浓度作用的黑素细胞与对照组相比,胞内可见较多呈束状的应力纤维,并集中分布于细胞膜内侧和细胞核周围,胞内荧光强度均高于对照组(P＜0.01).结论女贞子乙醇提取物及其单体酪醇在体外可促进黑素细胞的黏附及迁移,其机制可能与诱导黑素细胞内肌动蛋白细胞骨架的聚合有关.
목적 탐토녀정자을순제취물급기단체락순대인표피흑소세포점부화천이적조절작용.방법 체외분리배양인표피흑소세포,XTT법측정세포증식정황,용경FN포피적배양판측정세포점부솔,Transwell미공막법관측세포천이정황,격광공취초현미경관찰경중약처리적세포내기동단백세포골가적결구분포,반정량분석세포내형광강도.결과 0.0375～0.6 mg/ml녀정자을순제취물균가촉진흑소세포재FN상적점부(P＜ 0.05혹P＜ 0.01);0.5～2 mmol/L락순가촉진흑소세포점부(P＜0.05혹P＜0.01).0.15 mg/ml녀정자을순제취물급2 mmol/L락순무명현세포독성,재24 h시불명현촉진세포증식,선택차농도위공작농도,녀정자을순제취물급락순천과미공려막적흑소세포수분별위43.7화51.0개,현저고우대조조적20.3개(P＜ 0.01);량충약물공작농도작용적흑소세포여대조조상비,포내가견교다정속상적응력섬유,병집중분포우세포막내측화세포핵주위,포내형광강도균고우대조조(P＜0.01).결론 녀정자을순제취물급기단체락순재체외가촉진흑소세포적점부급천이,기궤제가능여유도흑소세포내기동단백세포골가적취합유관.
Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser microscopy was utilized to observe the structure and distribution of actin cytoskeleton in melanocytes,and cellular fluorescence intensity was determined by a semi-quantitative analysis.Statistical analysis was carried out by using unpaired t test.Results The adhesion activity of melanocytes to FN was significantly enhanced by the ethanol extract of 0.0375-0.6 mg/ml from glossy privet fruit (P ＜ 0.05 or 0.01),and by tyrosol of 0.5-2 mmol/L (P ＜ 0.05 or 0.01).As XTT assay showed,neither the ethanol extract of 0.15 mg/ml nor tyrosol of 2 mmol/L had cytotoxicity or promotive effect on cell proliferation.Hence,0.15 mg/ml and 2 mmol/L were determined as the working concentrations of ethanol extract and tyrosol respectively.The number of cells migrating through micropore membranes per high-power field (× 200) was 43.7 and 51.0 in melanocytes treated with the ethanol extract of 0.15 mg/ml and tyrosol of 2 mmol/L,respectively,significantly higher than that in untreated melanocytes (20.3,both P ＜ 0.01).Compared with untreated melanocytes,those treated with the ethanol extract of 0.15 mg/ml and those with tyrosol of 2 mmol/L showed higher intracellular fluorescence intensity (P ＜ 0.01) and more stress fiber bundles which congregated inside the cell membrane and around the nuclei.Conclusions The ethanol extract of glossy privet fruit and its monomer tyrosol can promote the adhesion and migration of human melanocytes in vitro,likely by promoting the congregation of actin cytoskeleton in melanocytes.