The Effect Of Bugan Jianyao Recipe On The Proliferation Of Rat Degenerative Endplate Chondrocytes And The Expression Of IKK? And NF-?B P65 MRNA

Objective:To observe the protective effect of Bugan Jianyao Decoction on rat degenerated endplate chondrocytes induced by interleukin-1? via the proliferation capacity and the expression of IKK? and NF-?B p65 mRNA,which may provide theoretical and experimental basis for the mechanism of Bugan Jianyao Decoction on IVDD.Methods:After the preparation of normal rat serum and Bugan Jianyao Decoction drug-containing serum,rat endplate chondrocytes were isolated,cultured,and verified by toluidine blue and collagen ? immunofluorescence staining.Experimental grouping and intervention were as follows: Blank group,induced group(0.5% volume fraction of IL-1?,10?g/L),normal serum group(induced group,with 5%,10% and 20% volume fraction of normal serum extra respectively)and Bugan Jianyao Decoction serum group(induced group,with 5%,10% and 20%volume fraction of drugcontaining serum extra respectively).Subsequently,Cell Counting Kit-8(CCK-8)assay was used to measure the proliferation of endplate chondrocytes at 24 h and 48h;real-time quantitative polymerase chain reaction(RT-q PCR)was used to detect I?B kinase?(IKK?)and NF-?B p65 mRNA expression.Results:Compared with the serum group and induced group corresponding to the volume fraction at 24 hours,the cell proliferation capacity increased,the expression of IKK? mRNA decreased in 48 hours(P < 0.05),distinctively,the expression of NF-?Bp65 mRNA was not changed(P> 0.05).Compared with the blank group,the proliferation capacity decreased and the expression of IKK?,NF-?B p65 mRNA increased in the induced group both at 24 h and 48h(P<0.05).Compared with the induced group(24 and 48h),the proliferation capacity increased after the treatment,while the expression of IKK?,NF-?B p65 mRNA decreased significantly(P < 0.05).In addition,significantly higher proliferation capacity and obviously lower IKK? and NF-?B p65 mRNA expression were observed in the 20% volume fraction of Bugan Jianyao Decoction serum group compared to 5% and 10% volume fraction Bugan Jianyao Decoction serum group(P<0.05).Conclusion:Bugan Jianyao Decoction can promote the proliferation capacity of rat degenerative endplate chondrocytes and regulate NF-?B pathway(inhibit the expression of IKK?,NF-?B p65 mRNA),which may be the potential mechanism of treating IVDD with the liver-reinforcing method.