结缔组织生长因子对NELL-1诱导的MC3T3-E1细胞成骨分化的影响

摘要/Abstract

摘要： 目的：探索结缔组织生长因子（CTGF）对尼克样1型蛋白（NELL-1）诱导的MC3T3-E1细胞成骨分化能力的影响。方法：以MC3T3-E1细胞为研究对象，分别使用PBS（对照）、NELL-1、CTGF和NELL-1+CTGF处理细胞，利用CCK-8法检测各组细胞增殖情况；成骨诱导后，利用茜素红染色和定量分析、磷酸酶（ALP）活性检测和ALP染色、实时荧光定量PCR和Western blot实验，检测各组细胞成骨分化情况。结果：与对照组相比，CTGF一定程度上抑制MC3T3-E1细胞增殖；NELL-1单独处理可诱导形成更多矿化结节，增强ALP活性，加深ALP染色，明显提高Runt相关转录因子2（RUNX2）、ALP、成骨细胞特异性转录因子Osterix（OSX）等骨形成基因的转录和蛋白表达水平（P<0.01）。与NELL-1单独处理组相比，NELL-1+CTGF联合处理组中细胞内矿化结节形成减少，ALP活性降低，ALP染色变浅，RUNX2、ALP、OSX等基因的转录和蛋白表达水平显著降低 (P<0.05)。结论：CTGF抑制NELL-1诱导MC3T3-E1细胞的成骨分化能力。

关键词: 结缔组织生长因子, 尼克样1型蛋白, 成骨作用

Abstract:

Objective: This study aims at exploring the role of connective tissue growth factor (CTGF) on Nel-1 like molecule type 1(NELL-1)-induced osteogenesis in MC3T3-E1 cells. Methods: The MC3T3-E1 cell lineage was used in the study, which was treated with each of control (PBS), NELL-1 alone, CTGF alone or in combination of NELL-1 and CTGF. Then cell proliferation was employed with help of CCK-8 assay. After osteogenic induction, multiple experimental techniques including alizarin red staining with quantitative analysis, ALP activity assay and ALP staining, quantitative real time poly chain reaction (qRT-PCR) and western blot were used to detect cell osteogenic differentiation among each group. Results: Compared to control, CTGF partly inhibited MC3T3-E1 cell proliferation; while NELL-1 treatment alone induced more mineralized nodules formation, promoted ALP activity and darker ALP staining in MC3T3-E1 cells with increased transcription and expression levels of osteogenic genes, such as RUNX2, ALP and OSX (P<0.01). However, compared to NELL-1 treatment alone, less mineralized nodules formation, decreased ALP activity and light ALP staining with dramatically down-regulated transcription and expression of RUNX2, ALP and OSX were found in cells treated with in combination of NELL-1 and CTGF (P<0.05). Conclusions: CTGF inhibited NELL-1-induced osteogenesis in MC3T3-E1 cells.s.?

Key words: connective tissue growth factor (CTGF), nel-like molecule type 1(NELL-1), osteogenesis

李彦秋李祖兵. 结缔组织生长因子对NELL-1诱导的MC3T3-E1细胞成骨分化的影响[J]. 口腔生物医学, 2023, (3): 169-174.

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