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Preptin Induces Human Osteoblasts Proliferation and Differentiation Through the ERK/CTGF Signaling Pathway

Author: ZhuJiaHua
Tutor: LiuYouShuo；YuanLingQing
School: Central South University
Course: Geriatrics
Keywords: Preptin Connective tissue growth factor Human osteoblasts Extracellular signal- regulated kinase Cell signaling pathways
CLC: R589.2
Type: Master's thesis
Year: 2010
Downloads: 98
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Abstract


Objective To study the Preptin on human osteoblast proliferation and differentiation , and to explore its mechanism. Methods primary cultured human osteoblasts based on the metabolic bone disease has been ruled out of normal adults by alkaline phosphatase (ALP), Ⅰ collagenase , the expression levels of osteocalcin and mineralized nodule formation of osteoblasts identified phenotype. Through the intervention group compared with the control group , observed Preptin stimulate osteoblast proliferation and differentiation of dose and time-dependent ; human osteoblast cell proliferation using [3H] thymidine incorporation assay , employing osteogenic differentiation of cell lysis protein determine the expression levels of alkaline phosphatase , spectrophotometer measurement p- nitrophenol to determine alkaline phosphatase activity. By detecting released into the culture medium of human osteoblasts connective tissue growth factor (CTGF), observe Preptin whether osteoblasts induced CTGF expression and its dose and time-dependent , CTGF by Western blot assay . Through small RNA interference (siRNA) inhibits the expression of CTGF osteoblasts , and Preptin before intervention with cell signaling blockers (PD98059, SP600125 or SB203580) pretreatment blocked human osteoblast mitogen-activated protein kinase (MAPK) signal transduction, in order to analyze whether CTGF and MAPK signaling pathways and how Preptin induced osteoblast proliferation and differentiation play a role. p38MAPK, extracellular signal -regulated kinase (ERK1 / 2), c-Jun N-terminal kinase (JNK) and its phosphorylation level of p-p38, p-ERK1 / 2, p-JNK by Western blot assay . Results Preptin can promote the proliferation of osteoblasts and increased alkaline phosphatase activity. Preptin be dose-and time -dependent manner regulate human osteoblasts CTGF expression through siRNA knockdown of human osteoblasts Preptin can reduce the expression of CTGF induced human osteoblast proliferation and differentiation. Preptin can induce osteoblast ERK phosphorylation of p38 and JNK no activation . In addition, human osteoblasts pretreated with ERK inhibitor PD98059 induced CTGF can Preptin reduced secretion and blocks Preptin of osteoblast proliferation and differentiation effects . Conclusion Preptin in human bone cells through ERK / CTGF cellular pathways regulating bone anabolic likely to be obese when one of the mechanisms of increased bone mass . CTGF is Preptin induced osteoblast proliferation and differentiation downstream regulated media.

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CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Metabolic diseases > Lipodystrophy
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