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Neuropeptide Y accelerates post-fracture bone healing by promoting osteogenesis of mesenchymal stem cells Xiao-chuan Gu, Xiao-Bin Zhang, Bing Hu, Ying Zi, Ming Li Neuropeptides Volume 60, Pages (December 2016) DOI: /j.npep Copyright © Terms and Conditions
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Fig. 1 Bone turn over markers in patients with simple or combined fractures. Serum samples were collected 1, 3, 7, 10, 14 and 28days after taking into hospital because of injuries. Then ALP, OC, PICP and ICTP levels were determined by ELISA. Fracture only: patients with simple tibial fracture. TBI fracture: patients with Traumatic Brain Injury (TBI) combined with tibial fracture. For ‘Fracture only’ group, data were collected from 20 patients at each time points, and for ‘TBI fracture’ group, data were collected from 60 patients at each time points. Data were analyzed by repeated measures AVONA and P values were calculated by post hoc scheffe test. Data are presented as mean±SD. *P<0.05. Neuropeptides , 61-66DOI: ( /j.npep ) Copyright © Terms and Conditions
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Fig. 2 NPY levels were higher in serums but not in CSF from patients with combined injuries than that in simple fractures. All Serum and CSF samples were collected at day 3 after injuries from patients with TBI fracture or fracture only. Fracture only: patients with simple tibial fracture. TBI fracture: patients with Traumatic Brain Injury (TBI) combined with tibial fracture. (A) NPY levels were determined by ELISA. Data were analyzed by repeated measures AVONA and P values were calculated by post hoc scheffe test. (B) NPY levels were determined by ELISA. Data were analyzed by One-way ANOVA and P values were calculated by post hoc scheffe test. (C) NPY levels were determined by ELISA. (D) NPY mRNA levels were determined by qPCR. Data are presented as mean±SD. *P<0.05, **P<0.01. Neuropeptides , 61-66DOI: ( /j.npep ) Copyright © Terms and Conditions
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Fig. 3 NPY promoted differentiation of osteoclasts in vitro. MSCs derived from health volunteers were differentiated in vitro for 7days. (A) CD34, CD73, CD105 and CD166 were detected by FACS. (B) ALP staining. 200×, Scale bar, 20μm. and (C) Alizarin Red S staining. 40×, Scale bar, 100μm. (D) and (E) NPY were added into the in vitro osteogenic differentiation system. Cbfa1 mRNA (C) and protein levels (D) were determined by qPCR and Western blot. Data were analyzed by One-way ANOVA and P values were calculated by post hoc scheffe test. Data are presented as mean±SD. *P<0.05, **P<0.01. Neuropeptides , 61-66DOI: ( /j.npep ) Copyright © Terms and Conditions
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Fig. 4 Y1 receptor was inducted by NPY administration and upregulated during osteogenic differentiation. (A) The mRNA levels of Y1 receptor were determined by qPCR during the in vitro differentiation of osteoblasts. (B) Total RNA were extracted from MSCs differentiated for 7days in the presence of different concentrations of NPY (0, 20nM and 50mM). The mRNA levels of Y1 receptor were determined by qPCR. Data are presented relative to the expression of GAPDH (mean±SD). Data were analyzed by One-way ANOVA and P values were calculated by post hoc scheffe test. ***P<0.001. Neuropeptides , 61-66DOI: ( /j.npep ) Copyright © Terms and Conditions
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