Affect Of Sodium Channel Blockers On Rabbit Articular Chondrocyte Cytoskeleton

Objective:Cytoskeleton morphology of rabbit chondrocytes being affceted of sodium channel blocker was observed by LCSM, and CSK protein fluorescence intensity of chondrocytes were determined, we wondered if there were differences between blocked and non-blocked chondrocytes in order to understand whether blockade of sodium ion channels will lead to morphological changes of the cytoskeleton, accordingly clarify mechanisms of articular cartilage degeneration from the cell level.Methods:2 Male New Zealand white rabbits aged 2 months, animals were killed by air embolism, pairs of knee chondrocytes under aseptic separation were in monolayer culture, the chondrocytes were divided into two groups namely, the intervention group and control group, the intervention group was added of 1mL DMEM culture medium containing 0.1mmol/L lidocaine, the control group was added of 1mL ordinary DMEM medium. The two groups of cells were placed in 37℃, saturated humidity,5%CO2 in the incubator, medium was changed every 3d, observed under inverted microscope, immunofluorescence fixation after two weeks, two groups of cartilage CSK proteins tubulin, actin and vinculin were fluorescent stained using immunofluorescence, CSK form of chondrocytes were observed using LCSM; fluorescence intensity of cartilage CSK protein was measured using the software.Results:Chondrocytes microtubules scattered radially from the nucleus throughout the cell and formed a loose and intertwined network structure throughout the cytoplasm, show uniform fluorescence intensity within the cytoplasm; intervention group formed more intertwined local microtubules than in the control group. Filamentous actin was distributed in the cytoplasm near the cell periphery Department, arranged to go along the cell contour lines, while no actin distribution in the nucleus; Spoted vinculin was distributed in the end of microfilaments near the cell membrane; actin presented thinning or even disappeared trend after intervention, while vinculin showed no significant change compared with intervention group.two groups of chondrocytes CSK quantitative determination of protein fluorescence intensity showed that tubulin fluorescence intensity values were 71.61±10.94,71.78±8.85 in the control group and intervention group, which showed no significant statistical difference (P>0.05); actin fluorescence intensity values of the control group and intervention group were 57.43±7.12,50.11±8.91, which showed significant statistical difference (P<0.05).Conclusion:CSK morphology of rabbit knee cartilage being blocked by the sodium channel blockers for two weeks showed significant changes comprared with unblocked ones; the intervention group cartilage actin protein content was statistically significant change compared with the control group; CSK morphology and protein levels both were both changed, indicating that the status of sodium ion channels was associated with the CSK morphology.