A purified extract from Clematis mandshurica prevents staurosporin-induced downregulation of 14-3-3 and subsequent apoptosis on rat chondrocytes.
A purified extract from Clematis mandshurica prevents staurosporin-induced downregulation of 14-3-3 and subsequent apoptosis on rat chondrocytes.
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DOI：
10.1016/j.jep.2006.11.012
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发表时间：
2007-05
影响因子：
5.4
通讯作者：
S. W. Lee;Sun-Mee Choi;Y. Chang;K. T. Kim;Tae Hyun Kim;H. Park;B. Park;You Jeong Sohn;S. Park;Su-Hyun Cho;W. Chung;Y. Yoo
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文献类型：
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作者：
S. W. Lee;Sun-Mee Choi;Y. Chang;K. T. Kim;Tae Hyun Kim;H. Park;B. Park;You Jeong Sohn;S. Park;Su-Hyun Cho;W. Chung;Y. Yoo
目的探讨东北铁线莲提取物对星形孢菌素诱导的大鼠关节软骨细胞凋亡的保护机制。结果东北铁线莲可阻止星形孢菌素诱导的几种抗凋亡bcl-2家族蛋白Bcl-xL和Bcl-2的下调，以及星形孢菌素诱导的凋亡bcl-2家族蛋白Bax的上调。东北铁线莲也防止staurosporin诱导的前线粒体抗凋亡蛋白14-3-3的下调。值得注意的是，针对14-3-3的siRNA取消了东北铁线莲对caspase-3激活的阻止。此外，细胞活力测定证实，沉默14-3-3基因取消了东北铁线莲的这种凋亡保护作用。免疫沉淀实验表明，东北铁线莲可抑制星形孢菌素诱导的14-3-3与phospho-ser 112-Bad和Bcl-xL相互作用降低为phospho-ser 155-Bad。
OBJECTIVETo dissect the mechanism of the protection of staurosporin-induced apoptosis on rat chondrocytes by a purified extract from Clematis mandshurica.DESIGNPrimary cultured rat articular chondrocytes as well as RCJ3.1C.18 cells were incubated with 1μM staurosporin and 300μg/ml purified extract from Clematis mandshurica. Western blot assay, silencing 14-3-3 gene and immunoprecipitation were conducted.RESULTSClematis 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.CONCLUSIONSClematis mandshurica prevents staurosporin-induced apoptosis of rat chondrocytes via 14-3-3.