Studies on mutagenic, antimutagenic and anti-inflammatory effects of water extracts from Chinese medicinal plants.

Abstract

This study was to explore the mutagenicities, antimutagenicities and anti-inflammatory effects of water extracts from Chinese medicinal plants, including Amomi fructus, Ampelopsis brevipedunculata (Maxim.) Trautv., Echinops grijsii, Millettia reticulata Benth, Rhinacanthus nasutus L. Kurz, Salvia plebeia R. Brown and Zizyphus jujuba Mill. The chemical compositions of water extracts from these prepared Chinese medicinal plants were also determined. The results showed that among the chemical compositions including moisture, protein, lipid, ash, carbohydrate and polyphenol each water extracts were different. All the water extracts (0.001~1000 μg/plate) demonstrated no mutagenicitiy against Salmonella typhimurium TA98 and TA100 whether cultured with or without S9 mix. However, S. plebeia (10 μg/plate) and A. fructus (1000 μg/plate) inhibited the mutagenicities of S. typhimurium TA98 induced with 4-NQO (0.5 μg/plate) by 50% or above. E. grijsii, A. brevipedunculata, S. plebeia and M. reticulata at the concentration of 1000 μg/plate inhibited the mutagenicities of S. typhimurium TA98 induced with IQ (50 ng/plate) by 50% or above. E. grijsii, R. nasutus, S. plebeia and A. fructus at the concentration of 1000 μg/plate inhibited the mutagenicities of S. typhimurium TA100 induced with 4-NQO (1 μg/plate) by 50% or above. E. grijsii, A. brevipedunculata, S. plebeia and M. reticulata at the concentration of 1000 μg/plate inhibited the mutagenicities of S. typhimurium TA100 induced with IQ (1 μg/plate) by 50% or above. To investigate anti-inflammatory effects of these water extracts from Chinese medicinal plants, pro-inflammatory cytokines, such as IL-1β, IL-6 and TNF-α, and anti-inflammatory cytokines, such as IL-10, secreted by peritoneal macrophages in vitro and in vivo from female BALB/c mice were determined. In vitro study, E. grijsii, R. nasutus, A. brevipedunculata, S. plebeia, Z. jujuba, A. fructus and M. reticulate at the concentrations of 500, 500, 250, 250, 500, 250 and 50 µg/mL, respectively, inhibited the inflammatory responses of peritoneal macrophages stimulated by lipopolysaccharide (LPS). In 28 days consecutive feeding study with female BALB/c mice, it was shown that E. grijsii, R. nasutus, A. brevipedunculata, S. plebeia, Z. jujuba, A. fructus and M. reticulate at the doses of 0.13, 0.08, 0.11, 0.07, 0.13, 0.04 and 0.06 g/kg body weight, respectively, also inhibited the inflammatory responses of peritoneal macrophages stimulated by lipopolysaccharide (LPS). The results showed that the seven Chinese medicinal plants extracts inhibited pro-inflammatory cytokines production (IL-1β, IL-6 and TNF-α) of peritoneal macrophages stimulated by lipopolysaccharide (LPS), while they increased anti-inflammatory cytokines IL-10 production. These results suggested that E. grijsii, R. nasutus, A. brevipedunculata, S. plebeia, Z. jujuba, A. fructus and M. reticulate might repress the inflammatory response induced by LPS in vitro and in vivo and demonstrate anti-inflammation potentials. These results also suggested that the water extracts from the seven medicinal plants might prevent and decelerate the injury during an inflammation status.

本論文旨在探討砂仁(Amomi fructus) 、山葡萄(Ampelopsis brevipedunculata (Maxim.) Trautv.)、山防風(Echinops grijsii)、雞血藤(Millettia reticulata Benth)、白鶴靈芝(Rhinacanthus nasutus L. Kurz)、荔枝草(Salvia plebeia R. Brown)及紅棗(Zizyphus jujuba Mill.)等七種中草藥水萃取物之致突變性、抗致突變性及對發炎反應的影響，利用體外(in vitro)及體內(in vivo)實驗模式，進行抗發炎功效之評估，此外，亦對此七種中草藥進行化學組成分析及基因毒性分析。 結果顯示，七種中草藥水萃取物之冷凍乾燥樣品成分之組成差異極大，除水分、灰分、蛋白質、脂質及醣類外，尚有大量多酚類存在。各水萃取物於所測試的濃度範圍(0.001~1000 μg/plate)內均對沙門氏菌變異株Salmonella typhimurium TA98、TA100不具致突變性。在抗致突變性方面，砂仁(1000 μg/plate)、荔枝草(10 μg/plate)可抑制4-NQO (0.5 μg/plate)對S. typhimurium TA98所誘發的基因突變50%以上，山葡萄、山防風、雞血藤及荔枝草於濃度1000 μg/plate時，可抑制IQ (50 ng/plate)對S. typhimurium TA98所誘發的基因突變50%以上；砂仁、山防風、白鶴靈芝及荔枝草於濃度1000 μg/plate時，山防風及雞血藤於濃度1000 μg/plate時，分別可抑制4-NQO (1 μg/plate)和IQ (1 μg/plate)對S. typhimurium TA100所誘發的基因突變50%以上。 於抗發炎研究中，若可抑制促發炎細胞激素(IL-1β、IL-6、TNF-α)或促進抗發炎細胞激素(IL-10)分泌，即判定具有抗發炎作用。在動物體外抗發炎實驗發現，砂仁、山葡萄、山防風、鷄血藤、白鶴靈芝、荔枝草及紅棗，在外在病原(lipopolysaccharide, LPS)存在下，可分別於培養濃度250、250、500、50、500、250及500µg/mL時，可抑制LPS (10 µg/mL)所誘導之腹腔巨噬細胞發炎反應。在動物體內試驗中亦發現，砂仁、山葡萄、山防風、鷄血藤、白鶴靈芝、荔枝草及紅棗，分別於管餵濃度0.04、0.11、0.13、0.06、0.08、0.07及0.13 g/kg body weight時，可抑制LPS (10 µg/mL)所誘導之腹腔巨噬細胞發炎反應。綜合體內及體外的實驗結果顯示，七種中草藥水萃取物皆可抑制LPS所誘導之發炎反應。動物體外試驗中以山防風、白鶴靈芝及荔枝草具有較佳的抗發炎效果。動物體內試驗中，以紅棗、荔枝草、鷄血藤及山防風具有較佳的抗發炎效果。