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目的研究滋补肝肾方复方及单方对小鼠 B-16黑色素瘤细胞酪氨酸酶 mRNA 表达的影响及滋补肝肾方的分子作用机制。方法定量逆转录 PCR 法(quantitative reverse transcriptional polymerase chain reac-tion,QRT-PCR)研究滋补肝肾方复方及单方对小鼠黑色素瘤酪氨酸酶 mRNA 表达的作用;mRNA 差别显示(mRNA differential display reverse transcriptional polymerase chain reaction,DDRT-PCR)法研究用滋补肝肾方复方及单方处理 B-16黑色素瘤细胞前后基因的差别表达。结果滋补肝肾方复方及各个单方并非都能够增加酪氨酸酶 mRNA 的表达量;但是滋补肝肾方复方和某些单方却能够激活小鼠黑色素瘤 B-16细胞线粒体 ATP合成酶-6(ATPase-6)基因的特异性表达。结论 QRT-PCR 结果表明不能将滋补肝肾方的治疗作用单纯解释为是酪氨酸酶表达提高的结果,而可能存在其他的作用机制,DDRT-PCR 结果表明 ATPase-6基因表达的激活可能是滋补肝肾方作用的关键环节。
Objective To investigate the effect of nourishing liver and kidney recipe and unilateral on the expression of tyrosinase mRNA in mouse B-16 melanoma cells and the molecular mechanism of nourishing liver and kidney. Methods Quantitative reverse transcriptional polymerase chain reac-tion (QRT-PCR) was used to study the effect of Zibu Ganshen recipe and unilateral on the expression of tyrosinase mRNA in melanoma of mice; mRNA differential display ( mRNA differential display reverse The transcriptional polymerase chain reaction (DDRT-PCR) method was used to study the differential expression of the gene before and after treatment with BZ-16 melanoma cells. The results showed that no tonic liver and kidney prescriptions and all prescriptions can increase the expression of tyrosinase mRNA; however, nourishing liver and kidney recipe and certain monotherapy can activate mouse melanoma B-16 cell mitochondrial ATP synthase-6 ( Specific expression of ATPase-6) gene. Conclusion QRT-PCR results show that the therapeutic effect of nourishing liver-kidney can not be simply interpreted as the result of increased expression of tyrosinase, and there may be other mechanisms of action. DDRT-PCR results suggest that the activation of ATPase-6 gene expression may be The key to nourishing liver and kidney function.