补肾强督治偻方对脂多糖刺激的小鼠RAW264.7细胞炎性细胞因子表达的影响

Effect of Bushen Qiangdu Zhilyu Decoction on Lipopolysaccharide-induced Inflammatory Cell Factors Expression in RAW264.7 Cells

【目的】研究补肾强督治偻方及其不同萃取成分对脂多糖(LPS)诱导的细胞炎性模型的细胞因子肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)表达的影响,探讨其主要的抗炎有效成分。【方法】采用系统溶剂法萃取补肾强督治偻方各成分,体外培养RAW264.7细胞,以LPS(1μg/m L)诱导炎症模型,补肾强督治偻方总方及不同溶剂萃取成分干预24 h,采用实时荧光定量PCR(QPCR)检测细胞TNF-α、IL-1βm RNA的表达。【结果】LPS刺激的RAW264.7细胞的炎症细胞因子TNF-α、IL-1βm RNA表达均显著升高(P<0.05),水与正丁醇萃取的补肾强督治偻方成分均可显著抑制RAW264.7巨噬细胞炎症模型TNF-α、IL-1βm RNA的表达(P<0.05),且呈一定的剂量依赖关系。【结论】水提取与正丁醇提取的补肾强督治偻方成分可能是其主要的抗炎有效成分部位,抑制炎症细胞因子基因表达可能是补肾强督治偻方治疗强直性脊柱炎的机制之一。

Objective To observe the effect of Bushen Qiangdu Zhilyu Decoction（ BQZD） and its different extracts on m RNA expression of tumor necrosis factor alpha（ TNF-α） and interleukin 1 beta（ IL-1β） in lipopolysaccharide（LPS）-stimulated RAW264.7 cells and to explore the effective anti-inflammatory constitutes.Methods Different constitutes of BQZD were obtained after extraction with ethyl acetate, n-butanol and water respectively. RAW264.7 cells were co-cultured with LPS in vitro to induce inflammatory cell model, and then were treated with BQZD and its different extracts for 24 hours. Real-time quantitative polymerase chain reaction（ RT-PCR） was used to measure m RNA expression of TNF-α and IL-1β in LPS-stimulated RAW264.7 cells.Results The m RNA expression levels of TNF-α and IL-1β were obviously increased in RAW264.7 cells after stimulated by LPS（P〈0.05）. The m RNA expression of TNF-α and IL-1β was significantly suppressed by BQZD water extract and n-butanol extract（P〈0.05）, in a dose-dependent manner. Conclusion Water extract and nbutanol extract may be the main effective anti-inflammatory constitutes of BQZD, and they can inhibit the m RNA expression of inflammatory cell factors, which may be one of the therapeutic mechanisms of BQZD in treating ankylosing spondylitis.