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

Effect of Danbai plastics anti-oxidantion and SCF/C-kit protein expression in cholasma rat model

[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Ha Cat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 m J/cm^2 UVB照射组、20 mg/mL丹白提取物组、30 mJ/cm^2 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/cm^2 UVB照射组、40 mg/mL丹白提取物组、30 mJ/cm^2 UVB照射+40 mg/mL丹白提取物组;Western blot法检测各组Ha Cat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。

[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.