动物造模 z-bio 2022-09-24 246

　　Objective To investigate the effect of estrogen deficiency on endplate chondrocyte degeneration.

　　Methods Forty 6-month-old rats were selected and divided into two groups: bilateral ovariectomy group (OVX) and sham operation control group (SHAM). 9 weeks after the operation, the materials were collected, and the cartilage endplate tissue was extracted and the primary endplate chondrocytes were cultured. The expression of collagen type II (COL-II) in the cartilage endplate tissue of SHAM group and OVX group was compared by immunohistochemistry. Inverted phase contrast microscopy and toluidine blue cell staining were used to identify endplate chondrocytes by observing cell morphology. The viability of endplate chondrocytes was compared between the two groups by CCK-8 method. Fluorescence staining of rhodamine-labeled phalloidin was used to observe the changes of F-actin in endplate chondrocytes after OVX. The changes of COL-II in endplate chondrocytes after estrogen deficiency were detected by cellular immunofluorescence. RT-qPCR method was used to compare the expression of SOX9, ACAN, ADAMTS-5, MMP13 and COL-X between the two groups.

　　Results Compared with the control group, the expression of COL-II protein in the cartilage endplates of the OVX group was decreased. Most of the endplate chondrocytes were polygonal and fusiform, arranged like paving stones. Endplate chondrocytes of OVX had decreased viability, more disordered cytoskeleton, increased stress fibers, decreased migration ability, and decreased COL-II expression. Compared with the control group, endplate chondrocytes in the OVX group had decreased expression of SOX9 and ACAN, and increased expression of MMP13, ADAMTS-5 and COL-X.

　　Conclusion Estrogen deficiency can cause endplate chondrocytes to degenerate. Endplate chondrocyte viability, migratory capacity, and bone synthesis decreased after OVX were accompanied by degradation of extracellular matrix and increased cell hypertrophic differentiation.