Inhibitory Effect Of Melittin On The Expression Of Matrix Metalloproteinases In Endplate Chondrocytes

Objective: To establish a degenerative model of natural passage of rat EPCs cells in vitro;The cytological characteristics and biological characteristics of EPCs in rats were observed;To compare the morphological and type II collagen expression of degenerative EPCs and normal EPCs in rats;Objective to study the inhibitory effect of Melittin on the expression of MMP in rat EPCs;Methods:1.The endplate cartilage of the lumbar spine of rats was obtained under aseptic condition and digested with collagenase type II to isolate the endplate cartilage of the intervertebral disc;2.Select P1 and P5 generation using inverted microscope to observe cell morphology,parallel toluidine blue staining and immunofluorescence staining were used for identification;3.MTT was used to determine the optimal concentration of Melittin for rat EPCs;4.Randomly selected P1 and P5 generation of cells in experimental groups: A.P1 blank control;the optimal concentration of B.P1 +Melittin;C.P5 control;D.P5 optimal concentration + Melittin;5 Cells in the four groups after the intervention by Western blot analysis(Western blot)to detect the expression of MMP-3.Result:1.The cultured EPCs gradually changed from polygonal to spindle;2.Toluidine blue staining of acidic Mucosubstance in the cytoplasm(such as proteoglycans)were stained dark blue,immunofluorescence staining of Col-2 cells,visible skeleton positive manifestation(green fluorescence),P5 generation is P1 Col-2 expression was significantly reduced,the above findings can prove that the cultured cells into chondrocytes;3.The optimum concentration of Melittin identified by MTT was1?g/ml;4.Melittin could inhibit the expression of MMP protein in degenerated EPCs,and can delay the endplate cartilage degeneration,the performance of the B,D is A,expression of C protein in MMP group significantly decreased the expression of B protein in MMP group than in D group significantly decreased,the difference was statistically significant(p<0.05).Conclusion: Melittin can inhibit the expression of MMP in EPCs,which has a certain protective effect of endplate cartilage,and provides some experimental evidence for the prevention and treatment of spinal degenerative diseases.