摘要
目的探讨黄芪甲苷对Th2型淋巴细胞株(D10.G4.1,D10)体外的干预作用。方法体外培养D10细胞,采用ConA作为刺激剂,分别予以2、4、8×10-5mol·L-1剂量黄芪甲苷体外干预,并用地塞米松作为对照。CCK-8法检测药物对D10细胞的增殖影响;流式细胞术检测药物对D10细胞凋亡和周期的影响;ELISA法检测药物对细胞培养上清IL-4、IL-5、IL-13及TNF-α水平的影响;Western blot法检测D10细胞gata-3表达水平。结果黄芪甲苷各剂量组对D10细胞增殖及凋亡均无明显影响(P>0.05);但黄芪甲苷高剂量组IL-13与gata-3表达水平明显下降(P<0.01),而低剂量组TNF-α明显下降(P<0.01)。结论黄芪甲苷体外具有抑制Th2反应的作用,但与药物细胞毒性或诱导凋亡无关。
Aim To evaluate the effects of astragalo-sideⅣon Th2 lymphocyte D10. G4. 1 (D10) in vitro.&amp;nbsp;Methods Cells were cultured in vitro according to ATCC recommendations and were treated with astraga-loside IV at dose of 2, 4, 8 &#215; 10-5 mol&#183;L-1 or dexam-ethasone ( Dex) in the present of ConA. Cell viability was determined by cell counting kit-8 assay. Apoptosis and cycle distribution of D10 were evaluated by flow cytometry. Th2 cytokines in the culture supernatant of D10 were evaluated by ELISA. Th2 transcription factor GATA-3 expression in D10 cells was determined by Western blot. Results Astragaloside IV at dose of 2, 4,8 &#215; 10 -5 mol&#183;L-1 have no significant effect on cell viability or apoptosis of D10 ( P〉0. 05 ) . Astragaloside IV at dose of 8 &#215; 10-5 mol &#183; L-1 markedly suppressed the level of IL-13 in the culture supernatant ( P 〈0. 01 ) , and the expression of GATA-3 ( P 〈0. 05 ) , and Astragaloside IV at dose of 2 &#215; 10 -5 mol&#183;L-1 ob-viously inhibited the level of TNF-α ( P 〈 0. 01 ) . Conclusion Astragaloside IV has immunomodulatory properties on Th2 response in vitro,which is not associ-ated with cytotoxic effect or apoptosis.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2014年第11期1508-1513,共6页
Chinese Pharmacological Bulletin
基金
国家重点基础研究发展计划(973计划)资助项目(No2009CB523001)
国家自然科学基金资助项目(No81403247)
浙江省医药卫生科技计划项目(No201473327)