加味桃红四物汤对黑素细胞酪氨酸酶和酪氨酸激酶受体基因蛋白表达的影响

Effect of Supplemented Four Agents Decoction Formula on Cell Prolifer ation and the Expression of Tyrosinase and c-kit Genes in Melanocytes In Vitro

目的研究加味桃红四物汤对体外培养的人表皮黑素细胞(MC)可溶性总蛋白以及酪氨酸酶(Tyr)基因和酪氨酸激酶受体基因(c-kit)表达的作用。方法采用免疫印迹法检测了加味桃红四物汤对酪氨酸酶和酪氨酸激酶受体蛋白含量的影响;595nm比色标准曲线测定黑素细胞总蛋白含量;采用细胞数统计和显微镜观察的方法测定加味桃红四物汤对细胞毒性和对细胞增殖的影响。结果加味桃红四物汤对黑素细胞可溶性总蛋白和酪氨酸酶的合成有促进作用,分别为4.76%和1.79%;对酪氨酸激酶受体蛋白的合成具有显著促进作用,高达79.41%。结论加味桃红四物汤治疗白癜风的作用机制之一是与酪氨酸激酶受体蛋白含量的提高相关,而与黑素细胞增殖和酪氨酸酶相关蛋白的含量无明显关系。

Objectives To study the effect of supplemented four agent s decoction formula (a decoction of Chinese medicines) on cell proliferation, ce ll toxicity and the expression of tyrosinase (Tyr) gene and c-kit (stem cell fac tor′s ligand/receptor) protein in melanocytes isoloated from Caucasians. Method s The expression of Tyr and c-kit protein was detected by Western blotting. Th e whole protein of melanocytes was measured by its absorption at 595 nm against the standard concentration curve. The toxicity and cell proliferation were obser ved by cell count and microscopy. Results The whole soluble protein content an d Tyr expression in cultured melanocytes were slightly increased 4.76% and 1.79% respectively after supplemented four agents decoction formula stimulation, as c ompared with those in the controls (P > 0.05). However, this medicine significan tly enhanced c-kit expression in melanocytes in vitro with an increase of 79.41% . Conclusions The efficacy of supplemented four agents decoction formula in th e treatment of vitiligo may be related to the SCF (stem cell factor)/c-kit pathw ay. This Chinese traditional medicine may enhance c-kit expression, but may not be associated with cell proliferation and Tyr expression of melanocytes.