Elucidation of Danzhixiaoyao Wan and Its Constituent Herbs on Antioxidant Activity and Inhibition of Nitric Oxide Production

Danzhixiaoyao Wan (DW) is a common 10 herbs formulation in China for regulating several clinical conditions affecting women. This research tried to explain one of DW's functions, purging heat, using in vitro pharmacological analyses. The whole formulation and each single herb of DW were compared based on antioxidant activity with the oxygen radical absorbance capacity (ORAC) assay, and for their inhibitory effect (IE) on nitric oxide (NO) production by lipopolysaccharide (LPS)-activated RAW 264.7 macrophages with the Griess assay. The results showed that DW as a whole formulation had both antioxidant activity and an IE on NO production, while the individual herb component of DW varied in their ORAC values and inhibition of NO production. The ORAC value of the whole DW was 450 μmol TE g−1. The order of antioxidant (ORAC) activity of the single herbs was: Mentha haplocalyx (1352 μmol TE g−1) > Glycyrrhiza uralensis (1184 μmol TE g−1) > Gardenia jasminoides (1129 μmol TE g−1) > Paeonia suffruticosa (465 μmol TE g−1), with the contributions being additive rather than synergistic. The production of nitrite by stimulated RAW 264.7 murine macrophages (unstimulated: 0.5 ± 0.1 μM versus LPS: 38.9 ± 2.3 μM) was significantly inhibited (P < 0.05) by M. haplocalyx, G. jasminoides, Bupleurum chinense and Paeonia lactiflora. DW as a whole had an IE on NO production, but this was not significant. The single herb M. haplocalyx had the highest ORAC value and the highest IE on NO production, followed by G. jasminoides. Both of these herbs have the ‘purging heat’ property in the theory of traditional Chinese medicine and this property of the samples may be correlated with the antioxidant activity and IE on NO production

Antioxidant activity of 45 Chinese herbs and the relationship with their TCM characteristics

Here, 45 Chinese herbs that regulate blood circulation were analyzed for antioxidant activity using the oxygen radical absorbance capacity (ORAC) assay. A recent publication by Ou et al. identified a close relationship between in vitro antioxidant activity and classification of Chinese herbs as yin or yang. The 45 Chinese herbs in this study could be assigned the traditional characteristics of natures (cold, cool, hot and warm), flavors (pungent, sweet, sour, bitter and salty) and functions (arresting bleeding, promoting blood flow to relieve stasis, nourishing blood and clearing away heat from blood). These characteristics are generalized according to the theory of yin and yang. We identified a broad range, 40–1990 µmol Trolox Equivalent/g herbs, of antioxidant activity in water extracts. There was no significant correlation between ORAC values and natures or functions of the herbs. There was a significant relationship between flavors and ORAC values. Bitter and/or sour herbs had the highest ORAC values, pungent and/or sweet herbs the lowest. Other flavors had intermediate values. Flavors also correspond with the yin/yang relationship and our results are supportive of the earlier publication. We reported for the first time antioxidant properties of many Chinese herbs. High antioxidant herbs were identified as Spatholobus suberectus vine (1990 µmol TE/g), Sanguisorba officinalis root (1940 µmol TE/g), Agrimonia pilosa herb (1440 µmol TE/g), Artemisia anomala herb (1400 µmol TE/g), Salvia miltiorrhiza root (1320 µmol TE/g) and Nelembo nucifera leaf (1300 µmol TE/g). Antioxidant capacity appears to correlate with the flavors of herbs identified within the formal TCM classification system and may be a useful guide in describing their utility and biochemical mechanism of action

Effects and potential mechanisms of Danzhi Xiaoyao Pill on proliferation of MCF-7 human breast cancer cells in vitro

Objective: To investigate the effects of 50% ethyl alcohol (EtOH) extracts from Danzhi Xiaoyao Pill (DXP) on the proliferation of MCF-7 human breast cancer cells and potential mechanisms. Methods: ATP-Lite assay was performed to test the proliferation of the MCF-7 breast cancer cell line; and antioxidant activity was measured by the oxygen radical absorbance capacity (ORAC). The effects of DXP on nitric oxide (NO) production were tested by lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages using the Griess reaction. Results: The 50% EtOH DXP extracts displayed a cytotoxic response on MCF-7 cells at 0.10, 0.25 and 0.50 mg/mL dose-dependently with the proliferation inhibited by more than 85%. The ORAC value of the DXP was 820 μ moL Trolox equivalent/g, about 40% of the vitamin C value. DXP extracts had significant inhibitory effect on NO production at the concentration from 0.0625 mg/mL to 0.5 mg/mL (P\u3c0.05, P\u3c0.01). Conclusion: The extracts of DXP could significantly inhibit the proliferation of MCF-7 cells, with the effect possibly related to its antioxidant activity and the inhibition of NO production

Effect of Honghua (Flos Carthami) on nitric oxide production in RAW 264.7 cells and α-glucosidase activity

Objective: To study the effects of extracts from Honghua (Flos Carthami) on lipopolysaccharide induced nitric oxide (NO) production in RAW 264.7 cells and the influence of the extracts on yeast α-glucosidase activity. The total flavonoid content of the extracts was also determined. Methods: Cytotoxicity of the extracts to RAW 264.7 cells was evaluated by the ATPlite™ method. Inhibitory effects of the extracts on NO production were evaluated by Griess assay. Curcumin was used as a positive control. Screening of extracts for potential α-glucosidase inhibitors was done by a fluorometric assay. The assay was based on the hydrolysis of 4-methylumbelliferyl-a-D-glucopyranoside to form the fluorescent product, 4-methylumbellifer-one. Acarbose was used as a positive control. The total flavonoid content was tested using kaempferol as the standard. Results: There were significant inhibitory effects on NO production when the extracts were 25-100 μg/mL (P\u3c0.05) and curcumin was 2-4 μg/mL (P\u3c 0.001). The extracts showed an inhibitory effect on α-glucosidase activity at the concentrations of 15.6-125 μg/mL with a half maximal (50%) inhibitory concentration (IC50) of (32.8 ± 5.7) μg/mL, compared with the IC50 of acarbose at (1.8±0.4) μg/mL. There was a significant difference between the two IC50 values (P\u3c0.001). The total content of flavonoids per gram of dried herb was 1.14 mg. Conclusion: Honghua (Flos Carthami) showed inhibitory effects on NO production in activated RAW 264.7 macrophage cells and an inhibitory effect on yeast α-glucosidase. There might be a relationship between these pharmacological effects and its flavonoid content