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目的观察六味地黄方对体外培养的人正常黑素细胞中ANNAT、HIOMT活性的影响。方法体外培养人正常黑素细胞,CCK-8法测药物法测定不同浓度的六味地黄方含药血清对黑素细胞增殖的影响,实时荧光PCR定量法测定不同浓度的六味地黄方含药血清对黑素细胞中ANNAT、HIOMT活性的影响。结果六味地黄方对于黑素细胞的增殖药物剂量的增加先促进了黑素细胞的增殖,在中剂量达到最高(P<0.05),继而高剂量组药物抑制其生长(P<0.05)。AANAT活性在低剂量组及中剂量组表达下调,与低中剂量促进黑素细胞增殖情况一致。分析结果表明,六味地黄方体外对黑素细胞增殖与AANAT活性存在相关性,且差异有统计学意义(P<0.05)。但六味地黄方含药血清对黑素细胞各用药剂量组HIOMT的mRNA含量无明显变化。结论 AANAT的活性在六味地黄方体外对黑素细胞影响的过程中可能起到限制性调节作用。
Objective To observe the effect of Liuwei Dihuang Recipe on the activity of ANNAT and HIOMT in human normal melanocytes cultured in vitro. Methods Human normal melanocytes were cultured in vitro. The effects of different concentrations of Liuweidihuangfang containing serum on the proliferation of melanocytes were determined by CCK-8 assay. The effects of different concentrations of Liuweidihuang decoction Effect of ANNAT and HIOMT activity in melanocytes. Results Liuweidihuang recipe increased the proliferation of melanocytes first and promoted the proliferation of melanocytes, reaching the highest level (P <0.05), then the high dose group inhibited the growth of melanocytes (P <0.05). The activity of AANAT was down-regulated in low-dose and middle-dose groups, which was consistent with the promotion of melanocyte proliferation by low and medium doses. The results showed that there was a significant correlation between the proliferation of ATRA and the proliferation of AANAT in vitro (P <0.05). However, the content of HIOMT mRNA of Liuweidihuangfang serum did not change significantly for each dose of melanocytes. Conclusions The activity of AANAT may play a limiting role in the process of melanocytes in vitro.