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

作　　者：唐旭东[1] 张路[2] 杨秀鹏[1] 唐玉凤[3] 王德秀 TANG Xudong;ZHANG Lu;YANG Xiupeng;TANG Yufeng;WANG Dexiu(Department of Hematology,Xiyuan Hospital China Academy of Chinese Medical Sciences,Beijing 100091,China;School of Management,Beijing University of Chinese Medicine,Beijing 100029,China;Laboratory,Xiyuan Hospital China Academy of Chinese Medical Sciences,Beijing 100091,China;Graduate School,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中国中医科学院西苑医院血液科,北京100091 [2]北京中医药大学管理学院,北京100029 [3]中国中医科学院西苑医院检验科,北京100091 [4]北京中医药大学研究生院,北京100029

出　　处：《天津中医药》2019年第11期1121-1125,共5页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家中医药管理局中医药行业科研专项(201507001-13);国家自然科学基金青年项目(81303127);国家自然科学基金面上项目(81673819);中国博士后科学基金项目(2014M551002)

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1μmol/L),C组为雄黄(浓度1μmol/L)+低剂量靛玉红(浓度0.05μmol/L)组,D组为雄黄(浓度1μmol/L)+高剂量靛玉红(浓度50μmol/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。[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 1μmol/L),group C was realgar(concentration 1μmol/L)+high dose indirubin(concentration 50μmol/L),group D was realgar(concentration 1μmol/L)+low dose indirubin(concentration 0.05μmol/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-γgene 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-γgenes is reduced.When the concentration of indigo naturalis is high,it can cause the increase of RIP and IFN-γgenes expression,and the expression of IRF3,TRAF3 and IL-10 genes can be reduced,which can promote the production of IFN-γand enhance the killing ability of NK cells against tumor cells.With the increase of the concentration of indigo naturalis,the IFN-γgene 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

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 

分 类 号：R285.6[医药卫生—中药学]