丹皮酚对骨质疏松骨折大鼠骨代谢的影响

Effect of paeonol on bone metabolism in osteoporotic fracture in rats

摘要:

[目的] 探讨丹皮酚对骨质疏松性（osteoporosis, OP） 骨折大鼠骨代谢、核因子κB （nuclear factor-kappa B, NF-κB） /基质金属蛋白酶-9（matrix metalloproteinases, MMP-9） 信号通路的影响。[方法] 52 只SD 雌性大鼠随机分为假手术组、模型组、骨化醇组及丹皮酚组，每13 只。除了假手术组外，其他大鼠均切除双侧卵巢，10 周后所有动物再行建立股骨骨折手术处理，假手术组、模型组分别给予生理盐水；骨化醇组给予0.05 μg/kg 阿法骨化醇；丹皮酚组给予200 mg/kg 丹皮酚灌胃，1次/d，连续12 周。行BMD 与骨微CT 定量检测，并血清检测、髓标志物mRNA 和蛋白表达检测。[结果] 给药12 周后，BMD[(0.31±0.06) g/cm² vs (0.26±0.06) g/cm² vs (0.25±0.05) g/cm² vs (0.19±0.04) g/cm², P＜0.001]、骨小梁数[(5.5±0.6) mm^-1 vs (4.7±0.6) mm^-1vs (4.5±0.5) mm^-1 vs (3.0±0.4) mm^-1, P＜0.001] 和骨小梁厚度[(78.5±5.4) μm vs (72.3±4.7) μm vs (70.8±5.0) μm vs (46.9±3.8) μm, P＜0.001] 由高至低依次均为假手术组＞丹皮酚组＞骨化醇组＞模型组，差异有统计意义。血清检测方面，PINP、BGP 和ALP 由高至低依次均为：假手术组＞丹皮酚组＞骨化醇组＞模型组，差异有统计意义（P＜0.05）。而血清TNF-α 、IL-6 和IL-8 由低至高依次均为：假手术组＜丹皮酚组＜骨化醇组＜模型组，差异有统计意义（P＜0.05）。通道标志物检测方面，NF-κB p65 和MMP 的mRNA 以及蛋白表达由低至高低依次均为：假手术组＜丹皮酚组＜骨化醇组＜模型组，差异均有统计意义（P＜0.05）。[结论] 丹皮酚能明显改善OP 骨折大鼠骨密度、骨微CT 测量和血清骨代谢指标，下调NF-κB/MMP9 信号通路标志物的表达。

Abstract:

[Objective] To investigate the effects of paeonol on bone metabolism, involving nuclear faction-kappa B (NF-κB) / matrixmetalloproteinases-9 (MMP-9) signaling pathway, in osteoporotic fracture in rat. [Methods] A total of 52 SD female rats were randomly di-vided into the sham operation (SO) group, model group, calcitriol group and paeonol group, with 13 in each group. Except those in the SOgroup, all the rats underwent bilateral total ovariectomy (OVX), and all animals underwent operation to establish femur fracture with inter-nal fixation 10 weeks later. The sham operation group and model group were respectively given normal saline, while the calcitriol group wasgiven 0.05 μg/kg alfacalcitol, and the paeonol group was given 200 mg/kg paeonol intragastrically once a day for 12 weeks. The BMD andbone microCT were conducted, and serum makers, signaling pathway markers mRNA and protein expression were detected. [Results] At 12weeks after drug administration, the BMD [(0.31±0.06) g/cm² vs (0.26±0.06) g/cm² vs (0.25±0.05) g/cm² vs (0.19±0.04) g/cm², P＜0.001], thetrabecular number (Tb.N) [(5.5±0.6) mm^-1 vs (4.7±0.6) mm^-1 vs (4.5±0.5) mm^-1 vs (3.0±0.4) mm^-1, P＜0.001] and bone trabecular thickness(Tb.Th) [(78.5±5.4) μm vs (72.3±4.7) μm vs (70.8±5.0) μm vs (46.9±3.8) μm, P＜0.001] were ranked from high to low as follows: the SOgroup＞the paeonol group＞the calcitriol group＞the model group with statistically significant differences. In terms of serum detections, thePINP, BGP and ALP were ranked in descending order as the SO group＞the paeonol group＞the calcitriol group＞the model group with statisti-cally significant differences (P＜0.05), while the serum TNF-α, IL-6 and IL-8 were in the order from low to high: the SO group＜the paeonolgroup＜the calcitriol group＜the model group with statistically significant differences(P＜0.05). In terms of signaling pathway marker detec-tion, both the mRNA and protein expressions of NF-κB p65 and MMP ranked as follows: the SO group＜the paeonol group＜the calcitriolgroup＜the model group, which all were statistically significant (P＜0.05). [Conclusion] The paeonol does significantly improve bone mineraldensity, bone microCT measurement and serum bone metabolism markers, while down-regulate the expression of NF-κB/MMP9 signaling pathway markers in OP fractured rats in this study.