Methotrexate prevents knee intraarticular adhesion via endoplasmic reticulum stress signaling pathway

doi:10.3969/j.issn.2095-4344.2016.24.018

Abstract

Abstract:

BACKGROUND: The pathogenesis of knee intraarticular adhesion is yet unknown. Excessive proliferation of fibroblasts is considered to cause knee intraarticular adhesion.
OBJECTIVE: To study the preventive effects of methotrexate on knee intraarticular adhesion through fibroblast apoptosis induced by endoplasmic reticulum stress.
METHODS: The viability of the cultured fibroblasts treated with methotrexate (10-5-10-9mol/L) or PBS was determined after 24 hours. Fibroblast apoptosis was detected by Hoechst33342 staining. Endoplasmic reticulum stress- and apoptosis-related proteins, including cleaved-PARP, CHOP, Bax and Bcl-2, were determined by western blot assay. Eighteen healthy male New Zealand white rabbits were used to establish the knee intraarticular adhesion models, and equally randomized into three groups, and received topical application of 2 or 1 g/L methotrexate, or normal saline (control). The preventive effects of methotrexate on knee intraarticular adhesion and CHOP expression in scar tissue were observed.
RESULTS AND CONCLUSION: Methotrexate inhibited the proliferation and viability of fibroblasts in a dose-dependent manner. The number of apoptotic fibroblasts was significantly increased compared with control group. Protein expression of cleaved-PARP, CHOP, and bax was increased, while protein expression of bcl-2 was decreased with time. The animal experiment showed that preventive effects of 2 g/L methotrexate on knee intraarticular adhesion were superior to 1 g/L methotrexate treatment. CHOP expression in the scar tissue in the methotrexate groups was higher than the control group and that was higher in high-dose methotrexate group. Our results suggest that methotrexate prevents knee intraarticular adhesion via endoplasmic reticulum stress-induced fibroblast apoptosis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Cicatrix, Methotrexate, Endoplasmic Reticulum, Apoptosis

CLC Number:

R318

Chen Hui, Li Xiao-lei, Yan Lian-qi, Sun Yu, Liang Yuan, Zhao Shuai, Zhu Geng-yao, Dai Ji-hang, He Jun, Wang Jing-cheng. Methotrexate prevents knee intraarticular adhesion via endoplasmic reticulum stress signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(24): 3615-3620.

Figures/Tables 2

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