| Objective:Along with the growth of rabbits age,we will observe change laws of autophagy activity of endplate chondrocytes in the intervertebral disc for exploring the correlation of age growth and autophagy activity change of endplate chondrocytes.Methods:Divided rabbits into four groups:young group(2months),adult group(12months),middle age group(24months)and old group(36months),three each group only and not distinguish by male or female.Rabbit to death by air embolism,under aseptic samples taken in each rabbit spine HE staining and endplate cartilage cell culture;the extracted endplate chondrocytes by toluidine blue staining;each endplate cartilage taken cell climbing,then alcohol fixed,nuclear stained with hematoxylin,eosin staining cytoplasm,and finally resin sealing piece;for each endplate cells were cultured to two generations,then cells climbing and adherenting paraformaldehyde,37℃MDC staining15min,used356nm fluorescence microscope filter and545nm emission filter for observation and radiography;dynamic expression by Western blot in each endplate cartilage cells in rabbit LC3autophagy-related proteins.Results:Blue after cell by toluidine blue staining prove the existence of endplate cartilage cells;each group of disc tissue HE staining nucleus gradually break;HE staining of cells in each age,spindle cells occur gradually deformation and aging;each endplate cartilage cells decreased the number of autophagic vacuoles by intracellular fluorescence decreased;Western blot LC3protein found in the dynamic changes with age,LC3-Ⅰ and β-action essentially unchanged,while LC3-Ⅱ protein gradually reduced.Conclusion:With the growth of the age of the rabbit,the rabbit intervertebral disc tissue degenerating,cells aging,chondrocytes endplate MDC staining taken on decrease the number of bubbles autophagic cells,LC3-Ⅰ basically unchanged and LC3-Ⅱ protein gradually decreased,indicating autophagic activity decreased gradually aging. |
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