五种兔唇花属植物全草水提物体外抗炎活性

Anti-inflammatory effects of aqueous extracts of whole herbs from five Lagochius species in vitro

目的研究五种兔唇花属植物全草水提物体外抗炎活性。方法采用脂多糖(LPS)刺激RAW264.7巨噬细胞建立细胞炎症模型,分为空白对照组(PBS)、模型对照组[PBS+LPS(3μg·mL^(-1)1和药物组[LPS(3μg·mL^(-1))+药物各浓度组(25、50、100、200、400μg·mL^(-1)];药物为冬青叶兔唇花、毛节兔唇花、二刺兔唇花、阔刺兔唇花与硬毛兔唇花全草水提物。MTT法测定细胞活力,Griess法测定NO含量,ELISA法测定TNF一α及IL-6的含量,RT-PCR测定iNOS、TNF一α及IL-6 mRNA的表达;采用DPPH法评价抗氧化活性。结果药物组五种兔唇花属植物全草水提物在25~400μg·mL^(-1)对巨噬细胞的活力无明显影响,与空白对照组相比,无显著差异(P>0.05);且在此浓度范围内能不同程度抑制巨噬细胞分泌NO、TNF一α、IL-6且无明显细胞毒性。其中,毛节兔唇花水提物在50~400μg·mL^(-1)能明显抑制巨噬细胞iNOS、TNF-α、IL-6 mRNA的表达,与模型对照组相比表现出非常显著的差异(P<0.01)。五种植物全草水提物在5~100μg·mL^(-1)还呈现出浓度相关性的抗氧化作用,毛节兔唇花水提物的清除能力最强。结论五种兔唇花属全草植物水提物具有潜在的抗炎作用,毛节兔唇花的活性最强,可能与下调相关炎症因子的分泌、基因表达及抗氧化作用有关。

AIM To investigate in vitro anti-inflammatory effect of aqueous extract from whole herbs of five Lagochilus species. METHODS LPS- induced RAW264.7 macrophage was used as an experimental inflammatory model. The cells were divided into three groups： control group （PBS）, model control group （PBS ＋ LPS （3 μg.mL-1））, drug group （LPS （3 μg.mL-1） ＋ aqueous extracts （25, 50, 100, 200, 400 μg.mL-1））. The aqueous extract was separately extracted from the whole herbs of Lagochilus ilicifolius Bunge （DQY）, L. lanatonodus C.Y.Wu et Hsuan （MJ）, L. diacanthophyllus （Palll） Benth （EC）, L. platyacanthus Rupr （KC）, L. hirtus Fisch. et Mey. （YM） . The proliferation of cells was examined by MTY assay. The level of nitric oxide （NO） was analyzed using Griess reaction, tumor necrosis factor ct （TNF-α） and interleukin-6 （IL-6） were determined using sandwich ELISA, and mRNA expressions of iNOS, TNF-α and IL-6 were detected by RT- PCR. DPPH-screening was selected to evaluate the antioxidant activity. RESULTS The aqueous extracts from five Lagochilus species were not able to alter the viability of macrophage at 25 - 400 μg＇mL-1 without obvious difference （P 〉 0.05） compared with the control group, but could decrease the production of NO, TNF-α and IL-6 in different degree, and L. lanatonodus showed relatively strong effect with correlation of concentration, while its aqueous extracts at 50 - 400 μg.mL-1 could obviously inhibit the mRNA expressions of iNOS, TNF-α, IL-6 with obvious difference （P 〈 0.01 ） compared with the model control group. The result of the 1, 1- diphenyl- 2- picrylhydrazyl （DDPH） assay showed that aqueous extracts at 5 - 100 p,g ＂mL-t also exhibited antioxidant properties at different degrees with correlation of concentration, and the aqueous extract of L. lanatonodus exibit the strongest effect. CONCLUSION The aqueous extracts of whole herbs from five Lagochilus species possessed potent anti-inflammatory properties with L. lanatonodus showing the best activity, which may contribute to its downregulating the inflammatory cytokines pathway and also attributing to its free radical scavenging activity.