桃红四物汤含药血清对巨噬细胞的影响

Effects of Taohongsiwu decoction on macrophages

目的探讨桃红四物汤含药血清对巨噬细胞分泌TNF-α、TGF-β1功能的影响,阐述中药治疗激素性股骨头坏死的作用机制。方法普通级新西兰兔,♀♂各半,随机分为3组,每日灌服浓度为1 g.ml-1的桃红四物汤药液,高剂量组15 ml.kg-1、低剂量组7.5 ml.kg-1,对照组15 ml.kg-1的生理盐水,每日1次,连续灌胃2周;巨噬细胞以5×105接种于6孔板中,以坏死骨微粒制备模型组,并予以含药血清干预48 h,ELISA检测各组上清中TNF-α、TGF-β1的含量;实时荧光定量PCR(RT-PCR)检测各组巨噬细胞TNF-α、TGF-β1mRNA的变化。结果荧光定量RT-PCR和ELISA检测结果可见桃红四物汤含药血清作用下的巨噬细胞分泌的TNF-α的量较空白血清组降低,组间差异具有统计学意义(P<0.05);桃红四物汤含药血清作用下巨噬细胞分泌的TGF-β1在转录水平和蛋白水平较空白血清组均有明显的升高,组间差异具有统计学意义(P<0.05)。结论桃红四物汤含药血清能提高巨噬细胞TGF-β1的表达,抑制TNF-α的表达,进而促进激素性股骨头坏死中骨的修复。

Aim To investigate the effect of serum containing Taohongsiwu decoction（ST） on transforming growth facor β1（TGF-β1）and tumor necrosis factor α（TNF-α）which are secreted by macrophages and to further explore the mechanism of Chinese herbal medicine in treating avascular necrosis of femoral head.Methods Conventional-12 New Zealand rabbits,half males and half females,were randomly divided into 3 groups： control group,high-dose group,low-dose group.Within 2 weeks,the high-dose group were administrated with ST 15 ml·kg-1 per day,the low-dose group were administrated with ST 7.5 ml·kg-1 per day,the control group were administrated with sodium chloride15 ml·kg-1 per day.Macrophages（M） were seeded in flat-bottom 6-well plates at 5×105 cells/well.Under the stimulation condition of necrotic bone particles,M were exposed to different concentrations of ST 48 h.Then supernatant and cells were respectively collected to detect the variation of TGF-β1,TNF-α by ELISA and real-time PCR.Results ELISA and real-time PCR showed the production of TGF-β1 was high-regulated by different concentrations of ST compared with control group（P＆lt;0.05）;the production of TNF-α was down-regulated by different concentrations of ST compared with control group（P＆lt;0.05）.Conclusion Taohongsiwu decoction can enhance the level of TGF-β1 and suppress the TNF-α,then promote bone repair of avascular necrosis of femoral head.