淫羊藿总黄酮对骨水泥微粒刺激单个核细胞产生分泌型TNF-α及TNF-α转换酶mRNA表达的影响

Effects of Epimedium brevicornum on aseptic loosening of prothesis: its effects on secretion of TNF-α and expression of TACE-mRNA in human monocytes induced by PMMA cement particles

目的 探讨淫羊藿总黄酮对人工全髋关节无菌性松动的影响。方法 随机选择准备行人工关节置换术患者 2 1例。采集外周血 ,分离单个核细胞 ,然后将其分为两大组 ,即Ⅰ、Ⅱ组。Ⅰ组标本分成 5个亚组 :单个核细胞组 (对照组 )、单个核细胞 +骨水泥微粒组、单个核细胞 +骨水泥微粒 +淫羊藿总黄酮 (5mg L)组、单个核细胞 +骨水泥微粒 +淫羊藿总黄酮 (10mg L)组、单个核细胞 +骨水泥微粒 +淫羊藿总黄酮 (2 0mg L)组。Ⅱ组分为3个亚组 :单个核细胞 +骨水泥 +佛波脂组、单个核细胞 +培养液组、单个核细胞 +骨水泥 +淫羊藿总黄酮 (10mg/L)组。培养后 ,放免法测定Ⅰ组细胞上清液中肿瘤坏死因子 -α(TNF -α)含量 ,逆转录聚合酶链反应 (RT -PCR)技术检测Ⅱ组TACEmRNA表达。结果 单个核细胞 +骨水泥微粒组的TNF -α含量明显高于对照组 (P <0 .0 1) ;单个核细胞 +骨水泥微粒 +淫羊藿总黄酮的TNF -α含量明显低于单核细胞 +骨水泥微粒组 (P <0 .0 1) ;而不同浓度淫羊藿总黄酮组之间的差异无统计学意义 (P >0 .0 5 )。单个核细胞 +骨水泥微粒组TACEmRNA的表达呈强阳性 ,而单个核细胞 +骨水泥微粒 +淫羊藿总黄酮组TACEmRNA的表达明显减弱。结论 骨水泥微粒可刺激单个核细胞分泌TNF -α。

Objective To explore the mechanism of Epimedium brevicornum (EB) on aseptic loosenin of prothetic hip joint by studying its effects on secretion of TNF-α and expression of TACE-mRNA in human monocytes induced by PMMA cement particles. Methods Blood samples (14 ml each) taken from 21 fracture patients (with no prothetic joint implanted), were processed to get the mononuclear cells (MNLC) to perform the experiments in two big groups. GroupⅠ, to test the effect of EB on the secretion of TNF-α, was divided into 5 subgroups: No. 1, culture of MNLC only; No. 2, MNLC + PMMA (cement particles suspended in culture medium RPMI/1640); Nos. 3, 4, 5, MNLC + EB (5 mg/kg, 10 mg/kg, 20 mg/kg respectively). GroupⅡ, to test the effect on the expression of TACE mRNA, was divided into 3 subgroups: No.1, MNLC + PMMA + PMA; No. 2, MNLC + RPMI/1640; No. 3, MNLC + PMMA + EB (20 mg/L). Each group was incubated for 48 hours before the level of TNF and the expression of TACE mRNA were determined. Results The leve of TNF was much higher in subgroup No. 2 than in the others ( Nos. 1, 3,4,5, P<0.01), but there were no significant differences in TNF between the subgroups Nos. 3, 4, 5 (P>0.05). The expression of TACE mRNA was increased significantly when PMMA particles was added in subgroup No. 1, as compared with that in No. 2, where only RPMI/1640 was present; while the expression of TACE mRNA was decreased significantly by EB in No. 3. Conclusion PMMA particles can stimulate MNLC to secrete TNF, while EB downregulates the expression of TACE mRNA, inhibiting the secretion of TNF and the associated bone resorption and loosening of prothesis. The present experiment demonstrates Epimedium brevicornum may clinically contribute to the prevention and treatment of aseptic loosening of prothesis.