通痹灵对多克隆刺激剂作用下小鼠T细胞CD69表达影响的血清免疫药理评价

Serum immunopharmacological assessment of effects of Tong- Bi- Ling on the expres -sion of CD69 on murine T cells stimulated with polyclonal activators

目的：应用血清免疫药理方法探讨通痹灵对T细胞体外活化的影响，以进一步揭示通痹灵的免疫调节分子机理，为临床用药提供理论依据。方法：分离小鼠淋巴结细胞，分别与CHX、5％CHX血清、CsA、5％CsA血清、5％通痹灵血清预孵后加入多克隆刺激剂继续培养，总计培养24h后收获细胞，进行双色免疫荧光标记，以流式细胞术对T细胞的CD69分子表达情况进行分析。结果：5％通痹灵作用下ConA活化的T细胞CD69表达百分率为（54．67±6．69）％，与相应对照［（64．35 ±10．13）%］比较有显著差异（P<0．05）；药物作用下PDB活化的T细胞CD69表达的百分率为（80．99±6．18）%，与相应对照［（79．68±4．17）％］比较无显著差异（P>0．05）。结论：通痹灵含药血清选择性地抑制ConA刺激的T细胞活化，而对PDB刺激的T细胞活化无影响；推测抑制类风湿关节炎疾病启动与发展中异常的T细胞活化，是通痹灵治疗类风湿关节炎重要机制之一；。

Abstract Objective:To investigate the effects of Tong Bi Ling(TBL) on the T cells activation by polyclonal activators in vitro withmethods of Serum Immunopharmacology for the application of this drug clinically.Methods:After separation of the lymphocytes from lymphoidnodes of C57BL/6 mouse, these cells were preincubated with CHX, serum containing CHX, CsA, serum containing CsA and serum containingTBL respectively for 1 h, then further incubated with polyclonal activators (ConA or PDB). Harvesting the cells after whole incubation for24 h, the authors estimated the expression rates of CD69 on T cells by flow cytometry following two-color immunofluorescent staining.Results:The expression rate of CD69 on the T cells preincubated with serum containing TBL after the stimulation in response to ConA was (54.67± 6.69) %, which showed significant difference with the expression rate of the control groups[(64.35 ± 10. 13) % ] (P<0.05);Whereas the ex-pression rate of CD69 on the T cells preincubated with serum containing after the stimulation in response to PDB was (80.99±6. 18)%,which didn't shaw significant for with tha expression rate of the control groups [(79.68 ± 4. 17)% ] (P>0.05) .Conclusion:5%serum containing TBL showed inhibitory effects on the activation of T cells in response to stimulation of Con A except that of PDB. These datasuggested that the inhibitory effects on the abnormal activation of T cells during the pathgenesis and progression of rheumatoid arthritis may beone of the important mechanisms applied to the treatment of the disease.