The Effect Of Bugan Jianyao Recipe On The Expression Of Aggrecan And MMP-3 And The Expression Of IKK? And NF-?B P65 Protein In Degenerated Rat Endplate Chondrocytes

Objective:To observe the effect of serum containing Bugan Jianyao Decoction on the expression of aggrecan,MMP-3,IKK?,NF-?B p65 protein in the model of degeneration of chondrocytes induced by IL-1?,and to explore the regulatory mechanism of Bugan Jianyao Decoction on NF-?B signal pathway,so as to provide experimental basis for the study of therapeutic mechanism of Bugan Jianyao Decoction on degenerative diseases of intervertebral disc.Methods:Normal rat serum and Bugan Jianyao Decoction drug-containing serum were prepared,and the rat endplate chondrocytes were successfully isolated.The type ? collagen immunofluorescence staining and toluidine blue staining were used to identify the endplate chondrocytes and then cultured to the third generation for experimental grouping and serum intervention.The experiment was randomly divided into eight groups:blank group,degeneration group,normal serum group and Bugan Jianyao Decoction drug-containing serum group.Except for the blank group,the other groups were given IL-1?(10 ug/L)to induce degeneration,in which the serum concentration gradient of 5%,10%,20%volume fraction was set in the two groups.After 24h and 48h of serum intervention in each group,cell culture medium was taken,and supernatant was taken after centrifugation.The levels of Aggrecan and MMP-3 were detected by ELISA,and the levels of IKK? and NF-?B p65 protein expression were detected by Western-Blot.Results:Compared with 24 h,the expression of Aggrecan in normal ser um group and Bugan Jianyao decoction containing serum group incr eased at 48 h,the expression of IKKa and NF-?B p65 protein decr eased(P<0.05),the expression of MMP-3 did not change significan tly(P>0.05);Compared with the degeneration group,the expression of Aggrecan,MMP-3,IKK?,NF-?B p65 protein in the normal seru m group showed no significant changes(P>0.05),the expression of Aggrecan in the drug-containing serum group increased significantl y(P<0.05),and the expression of MMP-3,IKK?,NF-?B p65 prote in decreased significantly(P<0.05);Comparison of drug-containing se rum group with normal serum group in the same volume fraction,Aggrecan expression increased significantly(P<0.05),while MMP-3,IKKa,NF-?B p65 protein expression decreased significantly(P<0.05);In the drug-containing serum group,20%volume fraction increas ed significantly compared with 5%and 10%volume fraction of Ag grecan expression(P<0.05),while MMP-3,IKK?,NF-?B p65 protei n expression decreased significantly(P<0.05).Conclusion:Bugan Jianyao Decoction can promote Aggrecan expression in degenerated rat endplate chondrocytes and inhibit the expression of MMP-3,IKKa and NF-?B p65 proteins by regulating the NF-?B si gnaling pathway.Therefore,it delays the degeneration of rat endplat e chondrocytes,which may be one of the mechanisms of Bugan Jia nyao Decoction in treating and delaying intervertebral disc degenerat ion.