复方甘草酸苷制剂对脂多糖诱导小鼠RAW264.7细胞分泌炎症因子的调节作用

Regulating Effect of Compound Glycyrrhizin on LPS-induced Inflammatory Factor Release from Murine RAW 264.7 Cells

目的:研究复方甘草酸苷片剂和注射剂对脂多糖诱导小鼠巨噬细胞RAW 264.7分泌炎症因子的调节作用。方法:采用脂多糖诱导的小鼠巨噬细胞RWA264.7,建立体外炎症模型。取复方甘草酸苷片剂和注射剂,制备供试药液。分别通过,MTT、Griess和双抗体夹心ABC-ELISA等实验,测定在不同浓度的供试药液作用下RAW264.7细胞活力以及经脂多糖诱导后的RAW264.7细胞的一氧化氮、肿瘤坏死因子TNF-α及白介素IL-1β、-6和-10等炎症因子分泌量的变化。结果:复方甘草酸苷的2种制剂药液在0～300μmol.L-1浓度范围内对RAW264.7细胞活力无影响。复方甘草酸苷注射剂药液在75、150和300μmol.L-1浓度下对经脂多糖处理的RAW264.7细胞的一氧化氮分泌抑制率分别为13.8%、40.4%和53.8%,并致细胞的TNF-α分泌量降低29.8%、41.5%和52.1%,IL-1β分泌量降低39.5%、55.6%和69.6%,IL-6分泌量降低18.3%、29.5%和39.1%,但IL-10分泌量却提高了8.2%、23.8%和30.8%;而复方甘草酸苷片药液在相同浓度下对同样细胞的一氧化氮分泌抑制率分别为9.8%、27.1%和37.8%,致细胞的TNF-α分泌量降低16.6%、37.0%和48.4%,IL-1β分泌量降低28.1%、47.9%和57.9%,IL-6分泌量降低13.5%、19.9%和28.2%,IL-10分泌量提高了3.9%、14.8%和23.4%。复方甘草酸苷的2种制剂对细胞分泌炎症因子的调节作用与阳性对照药地塞米松相似,其中注射剂的作用强于片剂。结论:复方甘草酸苷可剂量依赖性地显著抑制脂多糖诱导小鼠RAW 264.7细胞产生促炎因子一氧化氮、TNF-α及IL-1β和-6并促进抗炎因子IL-10的表达,这可能为其抗炎作用机制。

Objective： To investigate the regulating effect of compound glycyrrhizin tablet（GLT） and injection（GLI） on lipopolysaccharide （LPS）-induced inflammatory factor release from murine RAW 264.7 cells. Methods： The in vitro inflammation model was established using LPS-induced RAW 264.7 cells. The test solutions were prepared with GLT and GLI. The change of RAW264.7 cell viability and the secretion level of inflammatory factors, NO, TNF-α, IL-1β, IL- 6 and IL-10, from LPS-treated RAW264.7 cells exposed to different concentrations of the test solutions were determined respectively using MTT, Griess and Sandwich ELISA assays. Results： The two preparations exhibited no effect on RAW264. 7 cell viability at 0-300μmol·L^-1concentrations. 13.8%, 40.4% and 53.8% inhibition on the secretion of NO, 29. 8%, 41.5% and 52. 1% reduction in the secretion of TNF-α, 39.5%, 55.6% and 69.6% reduction in the secretion of IL-1β, 18.3%, 29.5% and 39.1% reduction in the secretion of IL-6 and 8.2%, 23.8% and 30. 8% increase in the secretion of IL-10 respectively by 75,150 and 300 μmol·L^-1 GLI , as well as 9. 8%, 27. 1% and 37. 8% reduction in the secretion of NO, 16. 6%, 37. 0% and 48. 4% reduction in the secretion of TNF-α, 28. 1%, 47.9% and 57.9% reduction in the secretion of IL-1β, 13.5%, 19. 9% and 28.2% reduction in the secretion of IL-6 and 3.9%, 14. 8% and 23.4% increase in the secretion of IL-10 respectively by 75,150 and 300 p, mol. L-1 GLT , were determined in LPS-treated RAW264. 7 cell culture. The regulating effect of the two preparations on LPS-induced inflammatory factor release from RAW 264. 7 cells was similar to that of the positive control drug dexamethasone, with GLI being superior to GLT. Conclusion： GL could significantly inhibit LPS-induced production of proinflamma- tory factors, NO, TNF-α, IL-1β and IL- 6, and provoke antiinflammatory factor IL-10 expression in RAW264. 7 cells in a concentration-dependent manner, which is its potential anti-inflammatory mechanism.