Geng Yi-Wei, Wang Ya-Lan, Deng Rong, Fu Kai-Li, Deng Yan
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2018 May 20;38(5):630-634. doi: 10.3969/j.issn.1673-4254.2018.05.21.
To investigate the role of autophagy in the regulatory effect of Shufeng Huoxue Fumula (SFHXF) on the proliferation and melanin metabolism in cultured murine B16 melanoma cells.
B16 cells were treated with solutions containing 0.12, 0.25, 0.49, 0.98, or 1.96 mg/mL SFHXF preparations, rapamycin (an autophagy inducer), or rapamycin+SFHXF. The changes in the proliferation of B16 cells were assessed using MTT assay, and tyrosinase activity and melanin content in the cells were determined. The expressions of autophagy-related proteins P62, p-mTOR, LC3B, and beclin 1 in the cells were detected using Western blotting.
Compared with the blank control cells, treatments with SFHXF both in the presence and in the absence of rapamycin concentration-dependently inhibited the cell proliferation (P<0.05) and obviously increased tyrosinase activity and melanogenesis in B16 cells (P<0.05); 0.98 mg/mL SFHLF, rapamycin+0.98 mg/mL SFHXF, and 50 nmol/L rapamycin all significantly up-regulated the expressions of LC3B-II and beclin 1 and down-regulated the expressions of P62 and p-mTOR in the cells.
SFHXF can regulate melanin metabolism and enhance tyrosinase activity and melanogenesis through the autophagy pathway to inhibit the proliferation of B16 cells in vitro.
探讨自噬在疏风活血方（SFHXF）对培养的小鼠B16黑色素瘤细胞增殖及黑色素代谢的调节作用中的作用。
用含有0.12、0.25、0.49、0.98或1.96mg/mL SFHXF制剂、雷帕霉素（一种自噬诱导剂）或雷帕霉素+SFHXF的溶液处理B16细胞。采用MTT法评估B16细胞增殖的变化，并测定细胞中的酪氨酸酶活性和黑色素含量。采用蛋白质印迹法检测细胞中自噬相关蛋白P62、磷酸化雷帕霉素靶蛋白（p-mTOR）、微管相关蛋白轻链3β（LC3B）和贝林1的表达。
与空白对照细胞相比，无论有无雷帕霉素，SFHXF处理均浓度依赖性地抑制细胞增殖（P<0.05），并明显增加B16细胞中的酪氨酸酶活性和黑色素生成（P<0.05）；0.98mg/mL SFHLF、雷帕霉素+0.98mg/mL SFHXF和50nmol/L雷帕霉素均显著上调细胞中LC3B-II和贝林1的表达，下调P62和p-mTOR的表达。
SFHXF可通过自噬途径调节黑色素代谢，增强酪氨酸酶活性和黑色素生成，从而在体外抑制B16细胞的增殖。
