补肾中药肾骨安对去势小鼠终板退变的影响

Effect of kidney-tonifying traditional Chinese medicine Shen Gu An on endplate degeneration in ovariectomized mice

  • 摘要:
    目的  探讨补肾中药肾骨安预防去势小鼠终板退变的作用和潜在机制。
    方法  30只雌性C57BL/6J小鼠随机分为假手术组(Sham组)、去卵巢组(OVX组)和中药组(HB组),每组10只。Sham组行假手术,OVX和HB组行双侧卵巢摘除。造模5天后,HB组给予中药汤剂灌胃,Sham组和OVX组给予生理盐水灌胃。3月后获取L4-5脊柱节段,Micro-CT观察椎体及终板微结构,番红O-固绿染色观察终板病理学改变,抗酒石酸酸性磷酸酶染色(TRAP)染色观察破骨情况,免疫组化检测终板Col2、CD31表达量。
    结果  Micro-CT结果显示,OVX组椎体骨小梁稀疏变少,体积骨密度(BMDtv)、骨体积百分比(BV/TV)、骨小梁数量(Tb.N)降低,而骨小梁间距(Tb.Sp)增加(均P < 0.05),终板出现病理重塑,体积(TV)及孔隙率[Po.V(tot)]增加(均P < 0.05)。与OVX组比较,HB组椎体骨小梁结构改善,BMDtv、BV/TV、Tb.N增加,Tb.Sp降低(均P < 0.05),终板重塑被抑制,TV和Po.V(tot)均降低(均P < 0.05)。番红O-固绿染色显示OVX组终板发生骨化重塑,HB组终板结构改善。TRAP染色见OVX组中终板下骨破骨细胞增加,HB组破骨细胞减少。免疫组化结果显示,与Sham组比较,OVX组椎间盘及终板Col2的表达降低,CD31表达增加,在终板骨化重塑区最为明显;而与OVX组比较,HB组终板的骨化重塑和CD31表达明显被抑制,Col2表达增加。
    结论  OVX引起小鼠椎体骨质疏松及终板骨化重塑,HB可以抑制终板破骨、血管新生、促进Col2表达,改善椎体骨小梁结构及终板的病理重塑。

     

    Abstract:
    Objective  To investigate the protective effect of the kidney-tonifying traditional Chinese medicine Shen Gu An on endplate degeneration in ovariectomized mice and its potential mechanisms.
    Methods  Thirty female C57BL/6J mice were randomly divided into a Sham operation group (Sham group), an ovariectomy group (OVX group), and a herbal medicine group (HB group) , with 10 mice in each group. The Sham group underwent sham surgery, while the OVX and HB groups underwent bilateral ovariectomy. Five days after modeling, the HB group received intragastric administration of the herbal decoction, while the Sham and OVX groups received intragastric saline. Three months later, L4-5 spinal segments were harvested. Micro-CT was used to observe the microstructure of vertebral bodies and endplates; Safranin O-Fast Green staining was used to observe pathological changes of the endplates; TRAP staining was used to assess osteoclasts; and immunohistochemistry was used to detect the expression of Col2 and CD31 in the endplates.
    Results  Micro-CT results showed that in the OVX group, vertebral trabeculae became sparse and reduced in number, bone mineral density of total volume (BMDtv), bone volume/total volume (BV/TV), and trabecular number (Tb.N) decreased, while trabecular separation (Tb.Sp) increased (all P < 0.05); pathological remodeling of the endplates occurred, with increases in total volume (TV) and total porosity (Po.V(tot)) (both P < 0.05). Compared with the OVX group, the HB group showed improved vertebral trabecular structure, with increased BMDtv, BV/TV, and Tb.N, and decreased Tb.Sp (all P < 0.05); endplate remodeling was inhibited, and both TV and Po.V(tot) decreased (both P < 0.05). Safranin O-Fast Green staining showed ossific remodeling of the endplates in the OVX group, while endplate structure was improved in the HB group. TRAP staining showed increased osteoclasts in subchondral bone in the OVX group and reduced osteoclasts in the HB group. Immunohistochemistry showed that, compared with the Sham group, Col2 expression in the intervertebral disc and endplate was decreased and CD31 expression was increased in the OVX group, most prominently in areas of ossific remodeling of the endplate. Compared with the OVX group, ossific remodeling and CD31 expression in the endplate were significantly inhibited and Col2 expression was increased in the HB group.
    Conclusions  OVX induces vertebral osteoporosis and ossific remodeling of the endplates in mice. HB can inhibit endplate osteoclast activity and angiogenesis, promote Col2 expression, and improve vertebral trabecular structure and pathological remodeling of the endplates.

     

/

返回文章
返回