Effects of Dang-Gui-Bu-Xue-Tang, an herbal decoction, on iron uptake in iron-deficient anemia

Figures & data

Figure 1 Experimental outline for the ferritin formation assay in Caco-2 cells.

Abbreviation: DBT, Dang-Gui-Bu-Xue-Tang.

Table 1 Formula of the AIN-93G diet

Item	(g/kg)
Casein	200
Cornstarch	629.486
Soybean oil	70
Fiber	50
Mineral mix AIN-93G, Fe-freea	35
Vitamin mix AIN-93Gb	10
l-Cysteine	3
Choline bitartrate	2.5
Total	1,000

Notes:

a The AIN-93G iron-free mineral mix contains the following components (in g/kg mixture): calcium carbonate (357), potassium phosphate monobasic (157), tri-potassium citrate monohydrate (102), NaCl (74), K₂SO₄ (46), MgO (24), zinc carbonate (1.65), manganese carbonate (0.63), cupric carbonate (0.3), KIO₃ (0.01), sodium selenite (0.01025), chromium potassium sulfate 12 hydrate (0.275), ammonium paramolybdate 4 hydrate (0.00795), sodium meta-silicate 9 hydrate (1.45), lithium chloride (0.0174), boric acid (0.0815), sodium fluoride (0.0635), nickel carbonate (0.0318), ammonium vanadate (0.0066), and finely powdered sucrose (235).

b The AIN-93G vitamin mix contains the following components (in g/kg feed): thiamine HCl (0.6), riboflavin (0.6), pyridoxine HCl (0.7), nicotinic acid (3.0), d-calcium pantothenate (1.6), folic acid (0.2), d-biotin (0.02), cyanocobalamin (0.0025), retinyl palmitate (400,000 IU), dl-α (all-rac)-tocopheryl acetate (7,500 IU), cholecalciferol (100,000 IU), K-1 (0.075), and finely powdered sucrose (974.655).

Abbreviations: AIN, American Institute of Nutrition; IU, international unit.

Figure 2 Animal model of iron-deficiency anemia.

Abbreviations: AIN, American Institute of Nutrition; DBT, Dang-Gui-Bu-Xue-Tang; Hb, hemoglobin; SD, Sprague-Dawley; h, hours.

Table 2 Phytochemical compositions of the DBT extract

Item	Method	Equivalent	Concentration range (µg/mL)	Linear equation	r^2	Content (mg/g)
Total polyphenol	Folin–Ciocalteu	Gallic acid	3.906–500	y=0.0056x+0.039	0.9999	6.51±0.29
Total flavonol	Vanillin	Catechin	3.906–250	y=0.0134x+0.1161	0.9996	2.31±0.05
Total saponin	Sulfuric acid	Diosgenin	7.812–1,000	y=0.0025x+0.0767	0.9970	34.48±3.99
Total Fe^2+	ICP-MS	–	–	–	–	0.0145
Ferulic acid	HPLC-UV system	Ferulic acid	0.488∼500	y=59550x+47568	1.0000	30.83±0.29

Abbreviations: DBT, Dang-Gui-Bu-Xue-Tang; ICP-MS, inductively coupled plasma mass spectrometry; HPLC-UV, high-performance liquid chromatography with ultraviolet detector; r², coefficient of determination.

Figure 3 Ferritin protein formations by Caco-2 cells and after co-treatment with DBT at low and high concentrations of FeSO₄.

Notes: Bars represent the mean ± standard deviation of three independent experiments. Statistically significantly differs from cell-only group at *P<0.05; **P<0.001. Statistically significantly differs from the FeSO₄-only group at ^#P<0.05; ^##P<0.001.
Abbreviation: DBT, Dang-Gui-Bu-Xue-Tang.

Figure 4 Ferritin protein formations by Caco-2 cells with an interval between administration of DBT and FeSO₄.

Notes: Bars represent the mean ± standard deviation of three independent experiments. Statistically significantly differs from the cell-only group at *P<0.05; **P<0.001. Statistically significantly differs from the FeSO₄-only group at ^#P<0.05; ^##P<0.001.
Abbreviation: DBT, Dang-Gui-Bu-Xue-Tang.

Figure 5 The gain in Hb in DIA rats with an interval of 2 h between the administration of DBT and FeSO₄.

Notes: Bars represent the mean ± standard deviation of six rats per group. Statistically significantly differs from the blank group at *P<0.05; **P<0.001; ***P<0.0001. Statistically significantly differs from the FeSO₄-only group at ^#P<0.05.
Abbreviations: DBT, Dang-Gui-Bu-Xue-Tang; DIA, diet-induced anemia; Hb, hemoglobin; h, hours.

Figure 6 Ferritin protein formation by Caco-2 cells when treated with FeSO₄ in combination with ferulic acid.

Notes: Bars represent the mean ± standard deviation of three independent experiments. Statistically significantly differs from the cell-only group at *P<0.05. Statistically significantly differs from the FeSO₄-only group at ^#P<0.05.

Figure 7 Effects of FA and DBT on the colorimetric determination of FeSO₄ concentration.

Notes: Results are shown as the mean ± standard deviation of three independent experiments, each performed in triplicate.
Abbreviations: DBT, Dang-Gui-Bu-Xue-Tang; FA, ferulic acid.