【摘要】 背景:在皮肤和毛发色素形成机制中,未曾有甘油二酯激酶ζ(diacylglycerol kinaseζ,DGKζ)参与色素形成的相关报道。目的:探讨DGKζ、蛋白激酶CβⅡ(Protein Kinase CβⅡ,PKCβⅡ)在不同毛色小鼠背部皮肤中的表达、定位及其与毛色形成的相关性。方法:取2月龄白色、灰色和黑色小鼠各6只,去毛取背部皮肤,Real-time PCR检测小鼠背部皮肤中DGKζ、PKCβⅡmRNA表达;Western blotting检测小鼠背部皮肤中DGKζ、PKCβⅡ和p-PKCβⅡ蛋白表达;免疫组织化学ABC法染色检测DGKζ和p-PKCβⅡ表达。实验方案经山西医科大学动物实验中心伦理委员会批准。结果与结论:(1)DGKζ、PKCβⅡ在3种毛色小鼠背部皮肤中m RNA表达趋势一致,均为:灰色组>白色组;黑色组>白色组,其差异有显著性意义(P<0.05);(2)PKCβⅡ活性的p-PKCβⅡ/PKCβⅡ,其变化趋势与DGKζ在3种毛色小鼠背部皮肤中表达趋势一致:灰色组>白色组;黑色组>白色组,其差异有显著性意义(P<0.05);(3)DGKζ和p-PKCβⅡ在灰色和黑色小鼠背部皮肤毛囊的外根鞘、毛母质细胞及毛母质的黑素细胞呈阳性表达,而在白色小鼠背部皮肤毛囊的外根鞘、毛母质细胞及毛母质的黑素细胞呈阴性表达;(4)结果说明,DGKζ、PKCβⅡ在黑、灰和白色小鼠背部皮肤中的表达与定位提示它们可能参与毛色的形成。更多还原

【Abstract】 BACKGROUND: In the skin and hair pigmentation mechanism, diacylglycerol kinase ζ(DGKζ) has not been reported to participate in pigmentation. OBJECTIVE: To investigate the expression and localization of DGKζ and protein kinase C βⅡ(PKCβⅡ) in the mouse back skin with different coat colors and their correlation with the formation of coat color. METHODS: The back skin of 2-month-old mice with different coat colors(white, gray, and black, 6 mice for each color) was taken to detect DGKζ and PKCβⅡ mRNAs by real-time PCR, DGKζ, PKCβⅡ and p-PKCβⅡ proteins by western blot, and DGKζ and p-PKCβⅡ expression by immunohistochemistry. The study protocol was approved by the Animal Ethics Committee of the Animal Experimental Center of Shanxi Medical University.RESULTS AND CONCLUSION: DGKζ and PKCβⅡ mRNAs were both expressed in the back skin of mice with three kinds of coat colors, with its relative expression amount being: gray group > white group; black group > white group(P < 0.05). Both DGKζ and PKCβⅡ proteins were expressed in the back skin of mice with different hair coat colors. The ratio of p-PKCβⅡ to PKCβⅡ showed that the change of PKCβⅡ activity had the same tendency with DGKζ expression: gray group > white group; black group > white group(P < 0.05). Immunohistochemistry staining showed that DGKζ and p-PKCβⅡ were positively expressed in the outer root sheath of hair follicle, hair matrix cells and melanocytes in the hair matrix in the back skin of gray and black mice, while were negative in the hair follicles of white mice. According to the expression and localization of DGKζ, PKCβⅡ in white, gray and black mouse back skin, we can speculate that DGKζ and PKCβⅡ may participate in the formation of the coat color.更多还原