Mice and experimental groups. Male DBA1/J mice (26 mice; age, 8 weeks; weight, 30-40 g) were acquired from The Second Affiliated Hospital of Zhejiang Chinese Medical University (Hangzhou, China). The mice were housed in a 22˚C temperature controlled room with 50-60% humidity, a 12 h light/dark cycle, and free access to food and water. The mice were randomly divided into three groups: Control group (n=6), model group (n=10) and puerarin group (n=10). In the model and puerarin groups, DBA1/J mice were injected with 4 mg collagen antibody (clones D1, F10, A2 and D8 to collagen type II; Chemicon; EMD Millipore, Billerica, MA, USA). Mice with collagen antibody-induced arthritis in the puerarin group were injected with 100 mg/kg puerarin (chemical structure presented in Fig. 1; Sigma-Aldrich, St. Louis, MO, USA) in 20% propanediol via the tail vein for each day for 7 days. Mice in the control and model groups were injected with same volume of saline. Observational clinical scores was determined on a scale from 0 to 3, using a digital Vernier caliper (VWR International, Radnor, PA, USA) as follows: 0= normal paws, no swelling or redness; 1= swelling and/or redness in one digit or in the ankle; 2= swelling and/or redness in one or two digits and ankle, and 3= entire paw is swollen or red.
Puerarin
Manufactured by Merck Group
Sourced in United States, Sao Tome and Principe
Puerarin is a plant-derived compound that can be used as a laboratory reagent. It functions as an antioxidant and is extracted from the kudzu root. This compound has applications in biochemical and cell-based research.
Automatically generated - may contain errors
Lab products found in correlation
Daidzin, by Merck Group (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Geniposide, by Merck Group (4 mentions)
Genistein, by Merck Group (3 mentions)
Daidzein, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
Genistin, by Merck Group (3 mentions)
Formononetin, by Merck Group (2 mentions)
Dionex ultimate 3000 hplc system, by Thermo Fisher Scientific (2 mentions)
Apigenin, by Merck Group (2 mentions)
Kaempferol, by Merck Group (2 mentions)
Catechin, by Merck Group (2 mentions)
Rutin, by Merck Group (2 mentions)
Myricetin, by Merck Group (2 mentions)
Naringin, by Merck Group (2 mentions)
Glycitin, by Merck Group (2 mentions)
Ly294002, by Merck Group (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
45 protocols using puerarin
1
Collagen-Induced Arthritis Model in Mice
2
Quantification of Isoflavones and Antioxidant Evaluation
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Reference standards of daidzein, genistein, puerarin, formononetin, and biochanin A with a purity of ≥98% were purchased from Sigma Aldrich (St. Louis, MO, USA) and used without further purification. Methanol (99.9%, for HPLC gradient), formic acid (98.0–100%, Puriss., for meeting analytical specifications of DAC and FCC), acetic acid (99.8%, for HPLC), and acetonitrile (99.9%, HPLC) were purchased from Sigma Aldrich. Choline chloride (99%; pharmaceutical grade) was purchased from Acros Organics, Geel, Belgium. Citric acid (99%, food-grade) was purchased from Sigma Aldrich. Quercetin, gallic acid, and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich. Ethyl acetate and ethanol were purchased from Himreaktivsnab company, Ufa, Russia. All other reagents and chemicals used in this study were of analytical grade. An ultrasonic cleaner and laboratory centrifuge Elma PE-6926 with a 10 × 5 mL rotor were used for the extraction process. A spectrophotometer Shimadzu-UV 1800 (Chiyoda-ku, Tokyo, Japan) was used as an analytical tool for the evaluation of total polyphenol content and antioxidant activity. A hot oven (Dry Oven UN55, Memmert, Schwabach, Germany) was used to dry the samples. In addition, HPLC-ESI-HRMS was used to quantify isoflavone content.
3
Puerarin Ameliorates Diabetic Nephropathy in Mice
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8-week old eNOS-null male mice on a C57BL/6 background were purchased from The Jackson Laboratory (Bar Habor, ME). Diabetes was induced by intraperitoneal injection of freshly prepared streptozotocin (STZ) (Sigma-Aldrich, Saint Louis, MO) dissolved in 0.1 M citrate buffer, pH 4.5 at 50 mg/kg after 4–6 hours of food withdrawal for 5 consecutive days. Control mice were injected with sodium citrate buffer. Blood glucose was measured every week. 2 weeks after diabetes was confirmed in mice (blood glucose > 250 mg/dl), mice were given puerarin (Sigma-Aldrich, Saint Louis MO) dissolved in 5% DMSO by oral gavage at a dose of 20 mg/kg body weight/day, or 5% DMSO vehicle as control, for 8 weeks. Urine samples were collected every week until they were sacrificed. All experimental methods were performed in accordance with the approved guidelines as described in the Guide for the Care and Use of Laboratory Animals42 . All animal studies were approved by the Institutional Animal Care and Use Committee at the Icahn School of Medicine at Mount Sinai, New York, NY.
4
Chromatographic Analysis of Isoflavonoids
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For chromatographic and chemical analyses, high-performance liquid chromatography (HPLC)-grade methanol was purchased from Merck Co. (Darmstadt, Germany) Ethanol, methanol, chloroform, and benzene, all of analytical grade, were obtained from Pecking Chemical Reagents (Beijing, China). Isoflavonoid standards, daidzin, and puerarin used for assay were purchased from Sigma Chemical Co. (St. Louis, MO, USA) Genistin, puerarin, daidzin, and daidzein (purity >97%) standards were purchased from the National Regulatory Authority, the Ministry of Public Health, DPR of Korea, and used as standards for thin-layer chromatography (TLC) analysis and assay.
5
Puerarin and Sterile Titanium Particles
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Puerarin (purity ≥98.0%, MW: 416.38) was purchased from Sigma-Aldrich (St. Louis, USA). Ti particles were obtained from Johnson Matthey Chemical (MA, USA). The particles were washed in 70% ethanol and sterilized at 180 °C for 6 h. Ti particles were endotoxin-negative, as verified by a Limulus amebocyte lysate assay (Biowhittaker, USA) (Frellsen et al., 2016 (link)).
6
MTT Assay for Cell Viability
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A 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) kit was purchased from Promega (USA). MMP9, ERK1/2, and phosphorylated ERK1/2 (p ERK1/2) antibodies were purchased from Abcam (USA). An RNA extraction kit, reverse transcription kit, and quantitative polymerase chain reaction (qPCR) SuperMIX were purchased from TAKARA (Japan). Puerarin was purchased from Sigma (USA). Matrigel was purchased from BD (USA). The gene amplification instrument and fluorescence imaging system were obtained from Bio-RAD (USA). A microscopic imaging system (Leica, Germany) from Germany was also used.
7
Culturing BGC-823 Gastric Cancer Cells
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The BGC-823 human gastric cancer cell line was purchased from the Shanghai Cell Collection (Shanghai, China). The cells were maintained in DMEM (Gibco-BRL Gaithersburg, MD, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% antibiotic solution (penicillin 100 U/ml and streptomycin 100 g/ml; Beyotime, Shanghai, China) at 37°C in a humidified atmosphere of 95% air/5% CO2. Puerarin and 5-FU were obtained from Sigma-Aldrich (P5555, F6627; St. Louis, MO, USA) and were 98% pure by reverse-phase high-performance liquid chromatography.
8
Puerarin Treatment for Mouse Inflammation
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In the therapeutic treatment protocol (Fig. 1A ), mice were treated with puerarin (Sigma-Aldrich; Merck KGaA) via intraperitoneal injection for continuous 7 days (8 mg/kg for the low dose, 40 mg/kg for the middle dose and 200 mg/kg for the high dose) (11 (link),12 (link)). As a positive control, dexamethasone (1 mg/kg) (Sigma-Aldrich; Merck KGaA) was administered intraperitoneally from day 1 to day 7.
9
Puerarin Modulates Vascular Inflammation
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Puerarin (Fig. 1 , purity: ≥ 98%) and SB203580 were purchased from Sigma-Aldrich (St Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Invitrogen (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from Gibco (Grand Island, NY, USA). Interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and vascular cell adhesion molecule-1 (VCAM-1) enzyme-linked immunosorbent assay (ELISA) kits were purchased from eBioscience (San Diego, CA, USA). Cell Counting Kit-8 (CCK-8), nitric oxide (NO), endothelin-1 (ET-1), superoxide dismutase (SOD) and malonaldehyde (MDA) kits were purchased from Jiancheng Bioengineering Institute (Nanjing, China). BCA protein assay kit was purchased from Beyotime (Shanghai, China). Proliferating cell nuclear antigen (PCNA), total-p38 mitogen-activated protein kinase (t-p38 MAPK), phospho-p38 MAPK (p-p38 MAPK) and β-actin antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA), Bromodeoxyuridine (BrdU) cell proliferation detection kit was purchased from KeyGen BioTECH (Nanjing, China), BrdU secondary antibody was purchased from Abcam (Cambridge, UK).![]()
Chemical structure of Puerarin
10
Estrogen and Dexamethasone Compound Effects
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DEX and 17β-estradiol (E2) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Puerarin (dissolved in dimethyl sulfoxide (DMSO; molecular weight 416.38; purity >98%) was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). DEX and pueranrin were used in a concentration gradient between 10−5 and 10−10 M. The final concentration of DEX (10−5 M) and of E2 (10−5 M) was dissolved in ethanol, and Puerarin (10−5 M) was dissolved in DMSO (Sigma-Aldrich). The DEX, Puerarin and E2 were stored at −20°C.
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