Western blot检测TRAF3通路产物变化在青黄散治疗骨髓增生异常综合征的机理  

The Mechanism of Western Blot Detection of TRAF3 Pathway Protein Product under the Function of Qinghuang Powder in the Treatment of Myelodysplastic Syndrome

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,Xi Yuan Hospital Affiliated to China Academy of Chinese Medical Sciences,Beijing 100091,China;School of Management,Beijing University of Chinese Medicine,Beijing 100029,China;Clinical Laboratory,Xi Yuan Hospital Affiliated to 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：《光明中医》2018年第14期2026-2030,共5页GUANGMING JOURNAL OF CHINESE MEDICINE

基　　金：国家中医药管理局中医药行业科研专项(No.201507001-13);国家自然基金青年基金(No.81303127);国家自然基金面上项目(No.81673819);中国博士后科学基金(No.2014M551002)

摘　　要：目的应用Western blot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TRAF3通路蛋白产物的变化,揭示青黄散治疗骨髓增生异常综合征(MDS)的机理。方法人骨髓增生异常综合征细胞(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,Western blot检测IFN-γ、TRAF3、IL10、IRF3、RIP、GAPDH蛋白产物。结果 TRAF3在C组表达明显增多,D组明显减少。IRF3在C组表达明显增多,D组明显减少。RIP在C组表达明显增多,D组明显减少。IL-10在D组有明显增加,C组明显减少。但是IFN-γ在蛋白水平没有检测到表达。TRAF3、IRF3、RIP、IL-10和IFN-γ在A和B组表达无明显统计学差异。结论青黄散通过作用于高度甲基化的TRAF3基因,使TRAF3基因去甲基化而重新发挥信号转导作用,青黛浓度较低时通过TRIF信号转导途径(上调IRF3、RIP表达,下调IL-10表达)促进机体产生IFN-γ,增强NK细胞对肿瘤细胞的杀伤能力;青黛浓度较高时通过TRIF信号转导途径(下调IRF3、RIP表达,上调IL-10表达)促进机体产生IL-10,抑制Th1细胞介导的免疫应答和炎症反应,起负调控的作用。Objective With the application of Western blot detection on myelodysplastic syndrome cell（ MUTZ-1） for protein changes in the TRAF3 pathway under the function of Qinghuang powder to reveal its mechanism. Methods MUTZ-1 culture according to the principle that A was as the control group,B with realgar as realgar group,C with 1μmol/L realgar ＋ 0. 05μmol/L indirubin as low dose of indirubin group,D with1μmol/L realgar ＋ 50μmol/L indirubin as high dosage group. All 4 groups were treated with Western blot on detection of IFN gamma,TRAF3,IL10,IRF3,RIP and GAPDH protein. Results The expression of TRAF3 in C group was significantly increased,and in D group decreased significantly. The expression of IRF3 in C group was significantly increased,in D group decreased significantly. The expression of RIP in C group was significantly increased,in D group decreased significantly. IL-10 increased significantly in the D group,in C group significantly reduced. But IFN-γ at the protein level was not detected. In A and B group,all the above 5 items had no Statistical difference. Conclusion Qinghuang powder by acting on the hypermethylation of the TRAF3 gene,induced TRAF3 gene demethylation and replayed a role in signal transduction,indigo concentration was low in the TRIF signaling pathway（ IRF3,upregulation of RIP expression,down-regulation of IL-10） to promote the body to produce IFN-γ,enhance the killing ability of NK cells to tumor cells; as the concentration was high by TRIF signal transduction pathway（ IRF3,down-regulation of RIP expression and upregulate the expression of IL-10） whose ability was to promote the body to produce IL-10,inhibited the immune response and inflammation mediated by Th1 cells,played a negative regulatory role.

关 键 词：血证 骨髓增生异常综合征 TRAF WESTERNBLOT 

分 类 号：R551.3[医药卫生—血液循环系统疾病]