柚皮苷抑制骨关节炎软骨细胞凋亡实验研究

Naringin inhibits human chondrocyte apoptosis in osteoarthritis

目的研究柚皮苷在炎症环境下对人骨关节炎(OA)软骨细胞凋亡的影响和可能的作用机制。方法采用不同浓度(0、0.1、1、10、100 mg/L)柚皮苷对OA软骨细胞进行预处理2 h后,加入或不加入炎症因子混合液(白细胞介素-1β5 ng/ml和肿瘤坏死因子-α20 ng/ml)共作用24 h。首先检测柚皮苷对炎症环境下软骨细胞活性和凋亡的影响,然后检测软骨细胞在柚皮苷作用下的一氧化氮(NO)水平及半胱氨酸天冬氨酸特异性蛋白酶(caspase)-3、caspase-8活性变化情况。结果柚皮苷预处理后软骨细胞在炎症因子刺激下发生凋亡显著减轻,且其抑制细胞凋亡作用具有浓度依赖性。进一步分子生物学检测显示,柚皮苷对NO生成及激活的caspase信号转导通路具有明显抑制作用。结论柚皮苷能有效抑制人OA软骨细胞凋亡,其作用机制可能是柚皮苷阻断了NO产生并抑制了下游的caspase信号通路,有望应用于OA治疗新药的研发。

Objective To investigate the anti-apoptotic effects of naringin on human osteoarthritis （OA） chondrocytes treated by inflammatory cytokines and the underlying mechanism. Methods OA Chondroeytes were pretreated with or without different concentrations （0, 0. 1, 1, 10, 100 mg/L） of naringin for 2 h and then co-treated for 24 h in either cell culture medium or a mixture of interleukin-Ⅲ （5 ng/ml） and tumor necrosis factor-α （20 ng/ml） （IT）. Cell viability and apoptosis were examined and molecules mediating apoptosis, induding nitric oxide （NO）, caspase-3 and caspase-8 were assayed. Results Naringin pretreatment inhibited OA ehondrocyte apoptosis induced by IT in a concentration-dependent manner. Mechanistically, the anti apoptotie effects of naringin resulted from decrease of NO production and inhibition of caspase pathways. Conclusion These results suggest that naringin might he a novel therapeutic agent for OA heeause of its suppression of apoptosis in OA chondrocytes via inhibiting NO and caspase signaling.