A purified extract from Clematis mandshurica prevents staurosporin-induced downregulation of 14-3-3 and subsequent apoptosis on rat chondrocytes

• Published in: Journal of ethnopharmacology 2007-05, Vol.111 (2), p.213-218
• Main Authors: Lee, Sung Won, Choi, Sun Mee, Chang, Yun Suk, Kim, Kyung Taek, Kim, Tae Hyun, Park, Hwan Tae, Park, Bong Soo, Sohn, You Jeong, Park, Soo Kyung, Cho, Su Hyun, Chung, Won Tae, Yoo, Young Hyun
• Format: Article
• Language: English
• Subjects: 14-3-3; 14-3-3 Proteins - metabolism; Animals; Apoptosis; Apoptosis - drug effects; bioassays; Biological and medical sciences; Cartilage; Cartilage, Articular - cytology; Cell Culture Techniques; Cell Line, Transformed; Cell Survival - drug effects; Cells, Cultured; chemical constituents of plants; chondrocytes; Chondrocytes - cytology; Chondrocytes - drug effects; Clematis; Clematis - chemistry; Clematis mandshurica; Down-Regulation - drug effects; Femur Head - cytology; General pharmacology; Humerus - cytology; Medical sciences; medicinal plants; medicinal properties; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; plant extracts; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Rats; Rats, Sprague-Dawley; Staurosporine - pharmacology
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2006.11.012

To dissect the mechanism of the protection of staurosporin-induced apoptosis on rat chondrocytes by a purified extract from Clematis mandshurica. Primary cultured rat articular chondrocytes as well as RCJ3.1C.18 cells were incubated with 1 μM staurosporin and 300 μg/ml purified extract from Clematis mandshurica. Western blot assay, silencing 14-3-3 gene and immunoprecipitation were conducted. Clematis mandshurica prevented staurosporin-induced downregulation of several antiapoptotic bcl-2 family proteins Bcl-xL and Bcl-2, and staurosporin-induced upregulation of an apoptotic bcl-2 family protein Bax. Clematis mandshurica also prevented staurosporin-induced downregulation of a premitochondrial antiapoptotic protein 14-3-3. It is noticeable that siRNA to 14-3-3 abolished the prevention of caspase-3 activation by Clematis mandshurica. Furthermore viability assay corroborated that silencing of 14-3-3 gene abolished this apoptosis protection efficacy by Clematis mandshurica. Immunoprecipitation assay elucidated that Clematis mandshurica prevented the staurosporin-induced reduction of the interactions between 14-3-3 with phospho-ser112-Bad and Bcl-xL to phospho-ser155-Bad. Clematis mandshurica prevents staurosporin-induced apoptosis of rat chondrocytes via 14-3-3.