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Protective effect of the serum of Jinggu Bitong decoction on the rabbits chondrocytes’apoptosis induced by interleukin-1β
XIAO Long YUAN Hai-zhou ZHONG Qin HUANG Jian XIAN Hua HUANG Ying-ru*
Acta Anatomica Sinica ›› 2015 ›› Issue (3) : 342-347.
Protective effect of the serum of Jinggu Bitong decoction on the rabbits chondrocytes’apoptosis induced by interleukin-1β
Objective To investigate the effect 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 Eight adult male New Zealand rabbits were randomly divided into four groups which were 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. 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 36 hours in group A, with IL 1-β 12 hours in group B, group C, group D and group E. Then the chondrocytes were cultured with 10% blank serum 24 hours in group B, with 10% PNLK serum in group C, with 10% Duhuo Jisheng decoction serum 24 hours in group D and with 10% Jinggu Bitong decoction serum 24 hours in group E. Real-time quantitative PCR (Real-time PCR) and Western blotting were used 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 24 hours, for the expressions of Bcl-2’s mRNA and protein in group C, group D and group E was higher than group B, group D and group E was 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＜0.05). Conclusion The Jinggu Bitong decoction has a significant protective action to the rabbit chondrocytes’s apoptosis induced by IL-1β, and the possible mechanism is to promote the Bcl-2’s expression and control the Bax’s expression and control proportional of Bcl-2 and Bax.
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