title: [Effect of Erigeron Breviscapus on the expression of OPG/RANKL/RANK in osteoblasts and pre-osteoclasts in vitro].
abstract: OBJECTIVE
  To study the effect of Erigeron Breviscapus (EB) at different concentrations and different intervention time points on the mRNA and protein expression of OPG/RANKL/RANK in MG63 osteoblast-like cells and RAW264. 7 pre-osteoclast cells cultured in vitro, thus exploring roles EB played in bone rebuilding and its mechanisms.
  
  
  METHODS
  MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro. The 3rd passage cells were divided into the control group and different experimental groups. Total RNA and protein were respectively isolated from cells treated with different concentrations of EB (0, 0.001, 0.01, 0.1, and 1.0 mg/mL) for 48 h. Meanwhile, the protein was extracted from 0 and 1 mg/mL EB groups at 12, 24, and 48 h respectively. Expression of OPG mRNA and RANKL mRNA in MG63 osteoblast-like cells, and expression of RANK mRNA in RAW264.7 pre-osteoclast cells were detected by semi-quantitative RT-PCR. Expression of OPG protein and RANKL protein in MG63 osteoblast-like cells, and expression of RANK protein in RAW264. 7 pre-osteoclast cells were detected by Western blot.
  
  
  RESULTS
  Along with increased EB concentration, expression of OPG mRNA and protein in MG63 osteoblast-like cells was gradually lowered (P < 0.05) after 48-h intervention of EB, the expression of RANKL mRNA and protein in MG63 osteoblast-like gradually increased (P < 0.05); the expression of RANK mRNA in RAW264.7 pre-osteoclast cells increased (P < 0.05). But the expression of RANK mRNA was slightly lower in the 0.1 mg/mL EB group than in the 0.01 mg/mL EB group, and the expression of RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). After treatment with 1 mg/mL EB for 12, 24, 48 h, the expression of OPG protein in MG63 osteoblast-like cells gradually decreased as time went by (P < 0.05), and the expression of RANKL protein in MG63 osteoblast-like and RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). The expression of RANKL protein in RAW264.7 pre-osteoclast cells increased as time went by (P < 0.05).
  
  
  CONCLUSIONS
  EB could inhibit the expression of OPG in osteoblasts in a dose- and time-dependent manner, promote the expression of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast, indicating EB might play roles in promoting bone resorption.
authors: Department of Stomatology, Guangfo Hospital, Guangzhou Medical University, Guangdong 528251, China. 502891288@qq.com
authors: Chang-Geng
authors: CG
authors: Liu
authors: Department of Stomatology, Guangfo Hospital, Guangzhou Medical University, Guangdong 528251, China.
authors: Qi-Xian
authors: QX
authors: Luo
authors: Department of Stomatology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors: Tian-You
authors: TY
authors: Ling
authors: Department of Stomatology, Guangfo Hospital, Guangzhou Medical University, Guangdong 528251, China.
authors: Ye-Yue
authors: YY
authors: Mo
authors: Department of Stomatology, Guangfo Hospital, Guangzhou Medical University, Guangdong 528251, China.
authors: Zi-Li
authors: ZL
authors: Cheng
authors: Department of Stomatology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors: Sheng-Gao
authors: SG
authors: Huang
authors: Department of Stomatology, Guangfo Hospital, Guangzhou Medical University, Guangdong 528251, China.
authors: Hui
authors: H
authors: Mo
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D049919:Erigeron; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism*; D053244:Osteoprotegerin / Q000378:metabolism*; D053245:RANK Ligand / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000378:metabolism
chemical_list: D004365:Drugs, Chinese Herbal; D053244:Osteoprotegerin; D053245:RANK Ligand; D012333:RNA, Messenger; D053246:Receptor Activator of Nuclear Factor-kappa B; C505686:TNFRSF11A protein, human; C506030:TNFRSF11B protein, human; C505688:TNFSF11 protein, human