京尼平苷对硝普钠诱导软骨细胞凋亡影响的实验分析.docxVIP

Abstract Objective To observe effects of Geniposide on SNP (Sodium nitroprusside) -induced apoptosis of the primary cultured chondrocytes of rats in vitro on proliferation and apoptosis and protein expressions of type II collagen, then explore treatment mechanism of Geniposide on OA. Methods (1) The rat articular chondroeytes were isolated with the method of enzyme digestion, morphology visualized by hematoxylin and eosin staining (HE) and identified by Collagen II of immunicytochemistry. (2) RandomLy divide the second generation of chondrocyte cells through synchronized cultivating were involved in experiment, into control, model and Geniposide groups. (3) Chondrocyte proliferation was measured by MTT assay. (4) Flow cytometry (FCM) were adopted to observe cell cycle and apoptosis rate, and NO examination adopted nitrate reductase method and efleets of Geniposide on the SNP-induced apoptosis of chondrocytes and NO in the culture supernatants were observed. (5) RT-PCR (reverse transcript-polymerase chain reaction) and Western Blot were applied to detect the mRNA and protein expressions of type II collagen? Results (1) The primary culture system of rat articular chondroeytes was successfully built.(2) Geniposide could decrease the percentage of SNP-induced chondroeytes in Go/Gi phase and increased percentage in S phase and G2/M phase (P 0.05) ,the apoptosis of chondrocytes and the concentration of NO in the culture supernatants were both reduced significantly (P 0.01 ) by MTT and FCM assay. (3) Geniposide intervention on SNP-induced apoptosis of chondrocytes was able to enhance both of mRNA and protein expressions of type II collagen. Conclusion Effects of Geniposide on SNP-induced apoptosis of chondrocytes were attributed to Geniposide could restrain the impacts of SNP on apoptosis of chondrocytes and thereby protect chondrocytes through regulating cell cycle, promoting proliferation, decreasing the apoptosis of chondrocytes and the concentration of NO in the cultur