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丹白涂膜剂对黄褐斑大鼠模型抗氧化作用及SCF/C-kit蛋白表达的影响
郭岱炯1, 孙佳瑜2, 陈宝清1, 杨玉峰1, 郭樱1, 华南3
1.深圳市中医院, 深圳 518021;2.深圳市福田区中医院, 深圳 518033;3.武警北京总队第三医院药剂科, 北京 100141
摘要:
[目的] 观察丹白涂膜剂对肌肉注射黄体酮及紫外(UV)照射导致黄褐斑大鼠模型的皮肤抗氧化作用及对SCF/C-kit蛋白表达的影响。[方法] 将大鼠随机分成对照组、模型组、氢醌组、丹白涂膜剂组共4组。用肌肉注射黄体酮及辅助UV照射法建立黄褐斑大鼠模型。测定各组皮肤组织中谷胱甘肽过氧化酶(GSH-Px)、超氧化物歧化酶(SOD)、酪氨酸酶(TYR)、总抗氧化能力(T-AOC),以及丙二醛(MDA)、一氧化氮(NO)及大鼠皮肤病理学检查。用CCK-8试剂盒观察0、5、10、20、40、80 mg/mL浓度丹白提取物对HaCat细胞、黑素细胞的抑制情况,选取最佳抑制浓度,将HaCat细胞分为1640培养基组、30 mJ/cm2 UVB照射组、20 mg/mL丹白提取物组、30 mJ/cm2 UVB照射+20 mg/mL丹白提取物组,黑素细胞分为MeIM-2培养基组、30 mJ/cm2 UVB照射组、40 mg/mL丹白提取物组、30 mJ/cm2 UVB照射+40 mg/mL丹白提取物组;Western blot法检测各组HaCat细胞SCF蛋白表达水平,各组黑素细胞的C-kit受体蛋白表达水平。[结果] 丹白涂膜剂能够显著增加黄褐斑模型大鼠皮肤中GSH-Px、SOD活性、T-AOC能力,降低MDA,抑制TYR增加;丹白提取物对HaCat细胞、黑素细胞的抑制情况呈剂量依赖性最大抑制效应分别为20 mg/mL(P<0.01)和40 mg/mL(P<0.01);丹白提取物能够下调UVB对HaCat细胞SCF蛋白和黑素细胞C-kit受体的促表达作用。[结论] 丹白涂膜剂对黄褐斑大鼠模型皮肤具有抗氧化作用,能够抑制与黄褐斑相关的蛋白SCF/C-kit蛋白表达。
关键词:  丹白涂膜剂  黄褐斑  抗氧化  SCF/C-kit蛋白表达
DOI:10.11656/j.issn.1672-1519.2017.10.16
分类号:R283
基金项目:广东省中医药强省科研基金项目(20141241)。
Effect of Danbai plastics anti-oxidantion and SCF/C-kit protein expression in cholasma rat model
GUO Dai-jiong1, SUN Jia-yu2, CHEN Bao-qing1, YANG Yu-feng1, GUO Ying1, HUA Nan3
1.Shenzhen Traditional Chinese Medicine Hospital, Shenzhen 518021, China;2.Shenzhen Futian District Traditional Chinese Medicine Hospital, Shenzhen 518033, China;3.Department of Pharmacy, The Third Hospital of Beijing Army Police, Beijing 100141, China
Abstract:
[Objective] To investigate the oxidation resistance and SCF/C-kit protein expression of the Danbai plastics in cholasma rat model induced by progesterone and ultraviolet radiation.[Methods] The rats were randomly divided into control group, model Group, hydroquinone group, Danbai plastics group.The cholasma model was establised with progesterone (7.5 mg/kg) by im injection orultraviolet radiation in female rat. The effects of each group on GSH-Px, SOD, tyrosinase (TYR), T-AOC, MDA, NO, in skin were detected and physical examination of rat skin. The HaCat cells and melanocytes inhibition was observed under the action of different concentrations of Danbai extract (0,5,10,20,40,80 mg/mL) by CCK-8. According to the suitable inhibition concentration,Hacat cells were divided into 1640 media group,30 mJ/cm2 UVB exposure group,20 mg/mL Danbai extract group,30 mJ/cm2 UVB exposure combined with 20mg/mL Danbai extract group. The melanocytes were divided into MelM-2 media group,30 mJ/cm2 UVB exposure group,40mg/mL Danbai extract group,30 mJ/cm2 UVB exposure combined with 40 mg/mL Danbai extract group. The SCF expressions of Hacat cells as well as the C-kit expressions of melanocyte were detected by Western bolt.[Results] The result showed that the groups with the Danbai plastics could signifcantly increase the activity of GSH-Px, SOD and T-AOC, decrease the contents of MDA and TYR in skin. CCK-8 results showed that HaCat cells and melanocytes proliferated in a concentration-dependent inhibition and HaCat cells inhibition reached a peak at the concentration of 20 mg/mL (P<0.01) and the melanocytes reached a peak at the concentration of 40 mg/mL (P<0.01). The results also showed that Danbai extract inhibited the SCF expression of Hacat and the C-kit expression of melanocyte and it reduced the promotion of SCF/C-kit expressions by UVB.[Conclusion] The Danbai plastics have anti-oxidantion and suppress SCF/C-kit protein expression.
Key words:  Danbai plastics  chloasma  antioxidation  SCF/C-kit protein expression
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