[甲状旁腺激素（1-34）对MG63细胞系中基质Gla蛋白表达及Wnt/β连环蛋白信号通路的影响]

[Effect of parathyroid hormone (1-34) on expression of matrix Gla protein and Wnt/β catenin signaling pathways in MG63 cell lines].

作者信息

Hu Ya-Li, Zhang Jie, Fu Liu-Chen, Yang Ya

机构信息

Department of Endocrinology and Metabolism, Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2016 Jun 20;36(7):984-9.

PMID:27435781

Abstract

OBJECTIVE

To observe the effect of parathyroid hormone (PTH)(1-34) on the expression of matrix Gla protein (MGP) and Wnt/β-catenin signaling pathway and elucidate the possible molecular mechanism of PTH (1-34) in the prevention and treatment of osteoporosis.

METHODS

MG63 cells treated with PTH (1-34) at 10(-9), 10(-8), and 10(-7) mol/L, alone or in combination with Wnt/β-catenin signaling pathway inhibitors DKK-1 (200 ng/ml) were examined for mRNA and protein expressions related with Wnt/β-catenin signaling with real-time PCR and Western blotting. The cell differentiation after the treatment was assessed with alkaline phosphatase (ALP) staining and cell viability assay.

RESULTS

PTH (1-34) significantly increased the expression of MGP in a dose-dependent manner in MG63 cells (P<0.05 or P<0.01). PTH treatment obviously enhanced ALP activity in the cells, and this effect was suppressed by DKK-1. Combined treatment with DKK-1 partially blocked PTH-induced enhancement of ALP activity (P<0.05). PTH promoted the expression of MGP and enhanced LRP5, β-catenin, and Runx2 expressions in Wnt/β-catenin signaling pathway at both protein and mRNA levels (P<0.05 or P<0.01). DKK-1 partially blocked the effect of PTH (1-34) on Wnt/β-catenin signaling pathway (P<0.05) without affecting MGP expression.

CONCLUSION

PTH (1-34) significantly increases the expressions of MGP and proteins in the Wnt/β-catenin signaling pathway. Wnt/β-catenin signaling pathway and MGP mediate the regulation of osteogenosis by PTH.

摘要

目的

观察甲状旁腺激素（PTH）（1-34）对基质Gla蛋白（MGP）表达及Wnt/β-连环蛋白信号通路的影响，阐明PTH（1-34）防治骨质疏松症的可能分子机制。

方法

用10⁻⁹、10⁻⁸和10⁻⁷mol/L的PTH（1-34）处理MG63细胞，单独处理或与Wnt/β-连环蛋白信号通路抑制剂DKK-1（200 ng/ml）联合处理，采用实时荧光定量PCR和蛋白质印迹法检测与Wnt/β-连环蛋白信号相关的mRNA和蛋白质表达。用碱性磷酸酶（ALP）染色和细胞活力测定评估处理后的细胞分化情况。

结果

PTH（1-34）能显著剂量依赖性增加MG63细胞中MGP的表达（P<0.05或P<0.01）。PTH处理明显增强细胞中的ALP活性，而DKK-1可抑制此作用。DKK-1联合处理部分阻断PTH诱导的ALP活性增强（P<0.05）。PTH在蛋白质和mRNA水平均促进Wnt/β-连环蛋白信号通路中MGP的表达，并增强LRP5、β-连环蛋白和Runx2的表达（P<0.05或P<0.01）。DKK-1部分阻断PTH（1-34）对Wnt/β-连环蛋白信号通路的作用（P<0.05），但不影响MGP表达。