钟航, 曹参, 杨静, 等.HIF-1信号通路与绝经后骨质疏松的关系研究.四川大学学报(医学版),2017,48(6):862-868
HIF-1信号通路与绝经后骨质疏松的关系研究
Research on Relationship of HIF-1 Signaling Pathway and Postmenstrual Osteoporosis
  
中文关键词:  去卵巢骨质疏松HIF-1α破骨细胞
英文关键词:OvariectomyOsteoporosisHIF-1αOsteoclast
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中文摘要:
      目的 初步探索低氧诱导因子-1(HIF-1)信号通路在绝经后骨质疏松(PMOP)发病过程中的作用及其作用机制。方法 C57BL/6J雌性小鼠随机分为假手术组(A组)、去卵巢绝经后骨质疏松组(B组)、溶剂对照组(C组)和HIF-1α抑制剂2-甲氧雌二醇(2ME2)治疗组(D组),每组15只。造模后3月处死小鼠,采用ELISA检测血清雌激素、小鼠Ⅰ型前胶原氨基末端肽(PINP)及小鼠Ⅰ型胶原羧基末端肽(CTX-1)等骨代谢指标, HE染色和抗酒石酸酸性磷酸酶(TRAP)染色观察骨结构及破骨细胞变化,免疫组化检测HIF-1α、HIF-1β、脯氨酰羟化酶结构域蛋白(PHD)、Hipple-Lindau肿瘤抑制蛋白(VHL)及低氧诱导因子抑制因子(FIH)等HIF-1信号通路的调节产物。取A、B组小鼠骨髓诱导培养破骨细胞,Western blot检测抑制蛋白激酶B (Akt)、细胞外调节蛋白激酶(ERK)、核因子-κB (NF-κB)信号通路后破骨细胞内HIF-1α变化。结果 B组小鼠骨代谢指标较A组升高( P<0.001),破骨细胞HIF-1α蛋白阳性表达,骨髓区HIF-1α蛋白表达量较A组升高( P<0.001),HIF-1β、PHD、VHL、FIH水平无明显变化。HIF-1α抑制剂抑制HIF-1后,去卵巢骨质疏松小鼠骨代谢指标降低( P<0.001),骨质疏松程度明显改善。予以Akt、ERK、NF-κB信号通路抑制剂干预后,去卵巢骨质疏松小鼠破骨细胞内HIF-1α水平均下降( P<0.05)。结论 HIF-1信号通路参与了小鼠PMOP的病理演变过程,抑制HIF-1信号通路可改善PMOP的严重程度;PMOP破骨细胞内HIF-1信号通路的上调可能与Akt、ERK、NF-κB这3条信号通路有关。
英文摘要:
      ObjectiveExploring the role of hypoxia-induceibal factor-1 (HIF-1) signaling pathway in postmenstrual osteoporosis (PMOP) and how to play a role in PMOP. MethodsSixty C57BL/6J female mouse were randomly divided into 4 groups: sham-operation group (A group), ovariectomized PMOP group (B group), HIF-1 alpha inhibitor 2-methoxy estradiol (2ME2) treatment group(D group) and solvent control group(C group), 15 mice in each group. There months after modeling, the metabolism product of mouse bone tissue including serum propeptide of typeⅠ procollagen (PINP), C-terminal telopeptide-Ⅰ (CTX-1) and serum estrogen were quantified by enzyme-linked immunosorbent assay (ELISA). Changes of bone structure were observed on HE stained tissue sections. Regulation products of HIF-1 signaling pathway including HIF-1α, HIF-1β, specific prolyl hydroxylases (PHD), Hipple-Lindau tumor suppressor protein (VHL), and factor inhibiting HIF (FIH) were measured by immuno-histochenmistry staining. Osteoclasts derived from OVX-mice were treated with inhibitors of extracellular regulated protein kinases (ERK), protein kinase B (Akt) and nuclear factor kappa B (NF-κB) signaling pathways. HIF-1α expression were detected by Western blot to obtain a rudimentary knowledge of possible mechanism of up-regulation of HIF-1α in osteoclasts of postmenospausal osteoporosis. ResultsMetabolism product of mouse bone tissue of B group were higher than A group ( P<0.001). The positive expression of HIF-1-alpha protein was found in osteoclasts, and the expression of HIF-1-alpha protein in bone marrow region was higher than that in A group ( P<0.001), while the change od HIF-1β, PHD, VHL, and FIH were not obviously. After the HIF-1α inhibitor treatment, markers of bone resorption of bone metabolism in ovariectomized mice reduced and the osteoporosis get greatly relieved ( P<0.001). HIF-1α expression was down-regulated in osteoclasts of OVX-mice after treated with inhibitors of ERK, Akt and NF-κB signaling pathways ( P<0.05). ConclusionHIF-1 signaling pathway get involved in the pathological evolution of osteoporosis after menopause, and inhibition of HIF-1 can significantly improve the severity of osteoporosis after menopause. ERK, Akt and NF-κB signaling pathways may involve in the up-regulation of HIF-1 signaling pathway in osteoclasts of PMOP.
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