Protective Effect Of The Serum Of Jinggu Bitong Decoction On The Rabbits Chondrocytes’ Apoptosis Induced By Interleukin-1β

Objective:To study the mechanism of osteoarthritis(OA), and probe into the effects of the Jinggu Bitong decoction on osteoarthritis(OA) proliferation and differentiation, by use serum of Jinggu Bitong (PNLK, drynaria rhizome, dipsacus asperoids and herba epimedii) decoction culture the chondrocytes in vitro, study the mechanism of the TCM therapy on osteoarthritis(OA) and the differentiation of chondrocytes by culture in vitro, in order to provide a platform to study the 1、degradation and restoration of osseous tissues. To study the effects and the mechanism of the Jinggu Bitong decoction on osteoarthritis (OA) by observing the effects of the serum of Jinggu Bitong decoction on the rabbits chondrocytes’ apoptosis and the expression of Bcl-2 and Bax.Methods:8 adult male New Zealand rabbits were randomly divided into four groups, and each group was gavaged with normal saline (NS), prescription for nourishing liver-kidney (PNLK, drynaria rhizome, dipsacus asperoids and herba epimedii), Duhuo Jisheng decoction and Jinggu Bitong decoction respectively. So the blank serum and the medicated serum were obtained. We identified chondrocytes by the toluidine blue staining and type II collagen immunofluorescent staining. The 3th passage rabbit chondrocytes were randomly assigned to 5 groups:blank group (group A), model group (group B), PNLK group (group C), Duhuo Jisheng decoction group (group D) and Jinggu Bitong decoction group (group E). The chondrocytes were cultured with 10% blank serum 36h in group A, with IL-1β 12h in group B, group C, group D and group E. Then the chondrocytes were cultured with 10% blank serum 24h in group B, with 10% PNLK serum in group C, with 10% Duhuo Jisheng decoction serum 24h in group D and with 10% Jinggu Bitong decoction serum 24h in group E. Use of Real-time quantitative PCR (RT-qPCR) and Western Blot to detect the expression of Bcl-2’s and Bax’s mRNA and protein. The apoptosis rate was analyzed by Flow cytometry (FCM).Results:All the cells were identified as chondrocytes. After the 24h, for the expressions of Bcl-2’s mRNA and protein,group C, group D and group E were higher than group B, group D and group E were higher than group C, and group E were higher than group D. The results were statistically different P<0.05. For the expressions of Bax’s mRNA and protein and the apoptosis rate, group C, group D, and group E were lower than group B, group D and group E were lower than group C, and group E were lower than group D. The results were statistically different P<0.05Conclusion:The serum of nourishing liver-kidney、Duhuo Jisheng decoction and Jinggu Bitong decoction all has the protective action to the rabbit apoptosis of chondrocytes, but the protection of Jinggu Bitong decoction is most significant, and the possible mechanism is acted by the mechanism maybe induced by IL-1β, the possible mechanism is acted by promote the Bcl-2’s expression and control the Bax’s expression and control proportional of Bcl-2 and Bax.