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目的:观察桃红四物汤含药血清对体外模拟骨坏死环境下的巨噬细胞的转化生长因子(TGF-β1)和肿瘤坏死因子(TNF-α)的mRNA转录水平的影响。进一步明确巨噬细胞在骨坏死修复中的机制。方法:普通级新西兰兔,雌雄各半,随机分为3组,每日ig 1 g.mL-1的桃红四物汤药液,高剂量组15 g.kg-1、低剂量组7.5 g.kg-1,对照组15 mL.kg-1的生理盐水,每日1次,连续ig 2周。U937经佛波醇肉豆蔻酸乙酸酯(PMA)诱导后的巨噬细胞以5×105接种于6孔板中,以坏死骨微粒制备模型组,并予以30%的含药血清干预48 h后收集细胞,RT-PCR检测各组巨噬细胞TGF-β1和TNF-αmRNA转录水平的变化。结果:桃红四物汤低、高剂量含药血清组TGF-β1mRNA的转录水平较空白血清组均趋呈现增高的趋势,组间差异具有统计学意义(P<0.05),其中低剂量为3.366 7,高剂量为4.186 7;TNF-αmRNA的转录水平较空白血清组则呈现减弱的趋势,组间差异具有统计学意义(P<0.05),其中低剂量为1.603 3,高剂量为1.356 7。结论:桃红四物汤可以提高坏死骨微粒刺激下的巨噬细胞TGF-β1mRNA水平,降低TNF-αmRNA水平,影响巨噬细胞的功能是其促进坏死骨修复的作用机制之一。
OBJECTIVE: To observe the effect of Taohongsiwutang medicated serum on the mRNA transcription of transforming growth factor (TGF-β1) and tumor necrosis factor (TNF-α) in macrophages stimulated by osteonecrosis in vitro. To further clarify the mechanism of macrophages in the repair of osteonecrosis. Methods: New Zealand white rabbits, male and female, were randomly divided into 3 groups. The rats were treated with 1 g.mL-1 of Taohong Siwu decoction daily, 15 g.kg-1 in high-dose group and 7.5 g in low-dose group. kg-1, control group 15 mL.kg-1 of normal saline once a day for two weeks. U937 macrophages induced by phorbol myristate acetate (PMA) were inoculated into 6-well plates at 5 × 105, and the model group was established with necrotic bone micro-particles, and 30% serum-containing intervention was performed for 48 h The cells were collected and the transcription level of TGF-β1 and TNF-α mRNA in macrophages were detected by RT-PCR. Results: The transcript levels of TGF-β1 mRNA in Taohong Siwu Tang low-dose and high-dose drug serum groups tended to increase compared with those in blank serum group, and the difference was statistically significant (P <0.05), among which the low dose was 3.366 7 , And the high dose was 4.186 7. The transcriptional level of TNF-αmRNA tended to be weaker than that of blank serum group. The difference between the two groups was statistically significant (P <0.05). The low dose was 1.603 3 and the high dose was 1.356 7. Conclusion: Taohong Siwu decoction can increase the level of TGF-β1 mRNA in macrophages stimulated by necrotic bone micro-particles, decrease the level of TNF-α mRNA and affect the function of macrophages.