摘要
目的：探讨白藜芦醇是否可通过抑制TLR4/NF-κB信号通路发挥抗骨性关节炎的作用。方法：采用两步酶消化法分离培养肥胖者骨性关节炎软骨细胞。免疫细胞化学染色及甲苯胺蓝染色对细胞进行表型鉴定。应用MTT法测定白藜芦醇对软骨细胞增殖活力的影响。ELISA法检测细胞培养基上清TNF-α的水平。RT-PCR法检测TLR4、NF-κBm RNA表达水平。结果：软骨细胞Ⅱ型胶原免疫细胞化学染色及蛋白多糖甲苯胺蓝染色呈阳性。白藜芦醇浓度在6.25~100μmol·L-1时对细胞增殖均无抑制作用,其中低浓度的白藜芦醇（6.25~12.5μmol·L-1）能促进细胞增殖（P〈0.05）。IL-1β（10 ng·ml-1）能抑制细胞增殖（P〈0.05）。白藜芦醇浓度在6.25~12.5μmol·L-1时,对IL-1β诱导的细胞活力降低有显著抑制作用。相比对照组,IL-1β诱导组的培养基上清中TNF-α水平及软骨细胞TLR4、NF-κBm RNA表达均明显增加（P〈0.01）,而加入白藜芦醇（6.25~12.5μmol·L-1）处理后,TNF-α水平及TLR4、NF-κBm RNA表达均显著降低（P〈0.01）。结论：白藜芦醇可能通过抑制TLR4/NF-κB信号通路发挥抗骨性关节炎效应。
Objective： To investigate whether resveratrol can exert anti-osteoarthritic effect via the TLR4 /NF-κB signaling pathway. Methods： Human articular chondrocytes were isolated and cultured by two-step eny matic digestion. The chondrocytes were identified by immunocytochemistry staining and toluidine blue staining. The cell viability was determined by the MTT assay. The levels of TNF-α in cell culture supernatant were measured by ELISA. The expression of TLR4 and NF-κB m RNA were examined by RT-PCR. Results： Cultured chondrocytes presented positive cells by Type Ⅱ collagen immunocytochemistry staining and toluidine blue staining. MTT assay showed that resveratrol（6. 25-100 μmol·L-1） had no discernable inhibition effects on chondrocytes cultured in the presence or absence of IL-1β. At 6. 25 and 12. 5 μmol·L-1,resveratrol significantly stimulated chondrocyte proliferation. In addition,viability of cells exposed to IL-1β（10 ng·ml-1） was significantly impaired but not in the presence of resveratrol（6. 25-12. 5 μmol·L-1）（P〈0. 05）. The level of TNF-α and the expression of TLR4 and NF-κB m RNA were significantly upregulated in the presence of IL-1β（10 ng·ml-1）（P〈0. 01） compared with control group,while resveratrol（6. 25-100 μmol ·L-1） treatment could dramatically downregulate TNF-αlevels and TLR4 and NF-κB m RNA expression（ P〈0. 01）. Conclusion： The anti-inflammatory effect of resveratrol on human articular chondrocyte may at least partly via the inhibition of the TLR4 / NF-κB signaling pathway.
出处
《东南大学学报（医学版）》
CAS
北大核心
2015年第3期342-346,共5页
Journal of Southeast University(Medical Science Edition)
基金
国家自然科学基金(81372971)
辽宁省自然科学基金(201202267)资助项目
