定量PCR检测TRAF3通路基因变化在青黄散治疗骨髓增生异常综合征中的机制

Mechanism of treatment on myelodysplastic syndrome with Q-PCR detection of TRAF3 gene pathway under the function of Qinghuang Powder

关键词:  骨髓增生异常综合征  肿瘤坏死因子受体相关因子  定量聚合酶链式反应

Key words:  myelodysplastic syndrome  TRAF  Q-PCR

摘要:

[目的]应用定量聚合酶链式反应（PCR）检测人骨髓增生异常综合征细胞（MUTZ-1）在明雄黄和靛玉红（青黛提取物）治疗后肿瘤坏死因子受体相关因子3（TRAF3）通路基因的变化，揭示青黄散治疗骨髓增生异常综合征（MDS）的机制。[方法]MUTZ-1培养，按A组为空白对照，B组为雄黄组（浓度1 μmol/L），C组为雄黄（浓度1 μmol/L）+低剂量靛玉红（浓度0.05 μmol/L）组，D组为雄黄（浓度1 μmol/L）+高剂量靛玉红（浓度50 μmol/L）组等4组进行处理，定量PCR检测IFN-γ、TRAF3、白介素-10（IL-10）、IRF3、RIP、GAPDH基因。[结果]TRAF3基因在B组表达较低，在C和D组表达较高，B、C和D组皆低于A组（P<0.05）。RIP基因在B组表达较高，在C和D组表达较低，B、C和D组皆高于A组（P<0.05）。IRF3基因在B和C组表达较低，在D组表达较高，B、C和D组皆低于A组（P<0.05）。IL-10基因在C组表达较高，在B组次之，在D组表达较低，B、C和D组皆低于A组（P<0.05）。IFN-γ在D组表达较高，在C组次之，在B组较低，D组高于A组（P<0.05）。[结论]青黄散（明雄黄和靛玉红的组合）在青黛浓度较低时可以引起RIP基因表达增加，TRAF3、IRF3、IL-10、IFN-γ基因表达减低，在青黛浓度较高时可以引起RIP、IFN-γ基因表达增加，IRF3、TRAF3、IL-10基因表达减低，都可以促进机体产生IFN-γ，增强NK细胞对肿瘤细胞的杀伤能力。随着青黛浓度增加，IFN-γ基因继续上调，IL-10继续下调，NK细胞对肿瘤细胞的杀伤能力继续增强。所以，未来可能通过调整青黛溶液的浓度，影响MUTZ-1人细胞株的TRAF3及其信号通路，从而达到良好的治疗目的。

Abstract:

[Objective] To reveal the mechanism of Qinghuang Powder in the treatment of myelodysplastic syndrome (MDS) by using Quantitative PCR to detect the change of TRAF3 pathway gene in human myelodysplastic syndrome cells (MUTZ-1) after treatment with realgar and indirubin (indigo naturalis extract).[Methods] MUTZ-1 was cultured according to group A as blank control group,group B was realgar group (concentration 1 μmol/L),group C was realgar (concentration 1 μmol/L)+high dose indirubin (concentration 50 μmol/L),group D was realgar (concentration 1 μmol/L) + low dose indirubin (concentration 0.05 μmol/L),and the IFN-γ,TRAF3,IL10,IRF3,RIP and GAPDH genes were detected by Q-PCR.[Results] The expression of TRAF3 gene in group B was low,in group C and D was high,and group B,C and D were lower than group A (P<0.05). The expression of RIP gene in B group was high,in group C and D was low,and group B,C and D were higher than group A (P<0.05). The expression of IRF3 gene in group D was high,in group B and C was low,and group B,C and D were lower than group A (P<0.05). IL-10 gene was highly expressed in group C,followed by group B and low in group D,and group B,C and D were lower than group A (P<0.05). IFN-γ gene was highly expressed in group D,followed by group C and low in group B,and group D were higher than group A (P<0.05).[Conclusion] Qinghuang Powder (combination of Ming realgar and indirubin) can increase the expression of RIP gene when the concentration of indigo naturalis is low,and the expression of TRAF3,IRF3,IL-10 and IFN-γ genes is reduced. When the concentration of indigo naturalis is high,it can cause the increase of RIP and IFN-γ genes expression,and the expression of IRF3,TRAF3 and IL-10 genes can be reduced,which can promote the production of IFN-γ and enhance the killing ability of NK cells against tumor cells. With the increase of the concentration of indigo naturalis,the IFN-γ gene continues to be up-regulated,IL-10 continues to be down-regulated,and the killing ability of NK cells to tumor cells continued to be enhanced. Therefore,in the future,it is possible to affect TRAF3 and its signaling pathway of the MUTZ-1 human cell line by adjusting the concentration of the indigo naturalis solution,so as to achieve a good therapeutic purpose.