黄精多糖对TNF-α作用的大鼠成骨细胞增殖、凋亡影响及其机制研究
Effect of polygonatum polysaccharide on proliferation and apoptosis of rat osteoblasts induced by TNF-α
  
DOI:10.3969/j.issn.1006-7108.2020.08.011
中文关键词:  黄精多糖  miR-212  成骨细胞  增殖  凋亡
英文关键词:polygonatum sibiricum polysaccharide  miR-212  osteoblasts  proliferation  apoptosis
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作者单位
刘超1 徐飞2 吴涛1 王子平3 赵晓明4* 1. 汉中市三二O一医院急诊科陕西 汉中 723000 2. 汉中市三二O一医院急诊科陕西 汉中 723000 3. 略阳县人民医院骨科 陕西 汉中 724300 4. 宝鸡市中心医院骨科 陕西 宝鸡 721008 
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中文摘要:
      目的 探讨黄精多糖(polygonatum sibiricum polysaccharide,PSP)对肿瘤坏死因子-α(TNF-α)作用的大鼠成骨细胞增殖、凋亡影响及其可能作用的机制。方法 采用MTT法检测不同浓度的PSP对TNF-α诱导的成骨细胞增殖能力的影响,筛选出PSP的适宜浓度;通过流式细胞术检测PSP对TNF-α诱导的成骨细胞凋亡能力的影响。采用实时荧光定量PCR(qRT-PCR)检测PSP对TNF-α作用的成骨细胞中miR-212表达的影响;运用蛋白印迹法(Western blot)检测成骨细胞中Cyclin D1、Bcl-2、Bax、P21的蛋白表达水平。结果 TNF-α可抑制成骨细胞增殖,促进P21蛋白表达,而抑制Cyclin D1蛋白表达。PSP不同剂量组可明显促进成骨细胞增殖,令Cyclin D1蛋白表达上调,而P21蛋白表达下调;TNF-α可促进成骨细胞凋亡,上调Bax表达,而降低Bcl-2、miR-212表达,PSP可提高miR-212的表达水平,上调miR-212表达可显著逆转TNF-α对成骨细胞的作用;抑制miR-212表达可明显逆转PSP对TNF-α诱导的成骨细胞增殖及凋亡的作用。结论 黄精多糖可通过上调miR-212表达,进而促进成骨细胞增殖及抑制细胞凋亡。
英文摘要:
      Objective To investigate the effects of polygonatum sibiricum polysaccharide (PSP) on the proliferation and apoptosis of rat osteoblasts induced by tumor necrosis factor-α (TNF-α) and its possible mechanism. Methods MTT assay was used to detect the effect of different concentrations of PSP on TNF-α-induced osteoblast proliferation, and the appropriate concentration of PSP was screened. Flow cytometry was used to detect the effect of PSP on the apoptotic ability of osteoblasts induced by TNF-α. Real-time quantitative PCR (qRT-PCR) was used to detect the effect of PSP on the expression of miR-212 in osteoblasts induced by TNF-α. The protein expression levels of Cyclin D1, Bcl-2, Bax and P21 in osteoblasts were detected by Western blot. Results TNF-α could inhibit the proliferation of osteoblasts, promote the expression of P21 protein, and inhibit the expression of Cyclin D1 protein. The different doses of PSP could promote the proliferation of osteoblasts, the expression of Cyclin D1 was up-regulated, and the expression of P21 was down-regulated. TNF-α could promote osteoblast apoptosis, up-regulate the expression of Bax, and decrease the expression of Bcl-2 and miR-212. PSP could increase the expression level of miR-212. Up-regulation of miR-212 expression could significantly reverse the effect of TNF-α on osteoblasts. Inhibiting the expression of miR-212 could significantly reverse the effect of PSP on TNF-α-induced osteoblast proliferation and apoptosis. Conclusion Polygonatum polysaccharide can promote the osteoblast proliferation and inhibit apoptosis by up-regulating miR-212 expression.
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