Hedgehog通路异常激活对小鼠关节软骨稳态的影响

Abstract

Abstract: Objective:To investigate the role of Hedgehog pathway in the mice osteoarthritis (OA). Methods:10-week-old male C57BL/6J mice were subjected to destabilization of the medial meniscus (DMM) surgery. The left hindlimb knee was the experimental group, with right hindlimb knee as control. Mice were sacrificed 8 weeks after the surgery and knee joints were harvested for subsequent experiments. Stereomicroscope was used to observe the damage degree of knee joint. Micro-CT was used to detect the meniscus calcification. Safranin O-Fast Green staining, toluidine blue staining, hematoxylin-eosin (HE) staining and immunofluorescence staining of Col2 were used to observe the changes of cartilage extracellular matrix and chondrocytes. The expression of genes related to articular cartilage homeostasis (Col2, Sox9, Mmp13) and Hedgehog signaling pathway (Smo, Gli1, Ptch1) were detected by RT-qPCR. ATDC5 cell model of OA was established using IL-1β, treated with NVP-LDE225 as Smo inhibition. RT-qPCR was used to detect the expression of Smo, Gli1, Ptch1, Col2, Sox9 and Mmp13. Results:Mouse OA model was established successfully after the DMM surgery. The experimental mice present knee joint enlargement, destroyed cartilage layer, higher meniscus calcification volume, knee cartilage deformation, decreased content of extracellular matrix proteoglycan and disordered arrangements of the chondrocytes. The immunofluorescence staining of Col2 indicated an aberrant extracellular matrix. In contrast, Col2 and Sox9 were significantly downregulated. Mmp13 and Hedgehog related genes (Smo, Ptch1, Gli1) was significantly increased in the OA model mice（P＜0.05）. Besides, inhibition of Smo in ATDC5 cell OA model antagonized the downgraded gene expression of Col2 and Sox9 and upgraded gene expression of Mmp13, Smo, Ptch1 and Gli1 induced by IL-1β, thus attenuating the severity of OA（P＜0.05）. Conclusions:Hedgehog signaling was activated in OA and Hedgehog signaling might be involved in the occurrence and development of OA.

Key words: osteoarthritis, cartilage, Hedgehog signaling

References 22

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Role of Hedgehog pathway in preserving integrity of the mouse articular cartilage

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