Human renal cancer 786-O cells were obtained from Shanghai Cell Institute Country Cell Bank (Shanghai, China). 786-O cells were grown in DMEM supplemented with 10% fetal bovine serum (FBS) (Gibson/BRL, MD, USA), 100 U/mL penicillin G, and 100 μg/mL streptomycin (Sigma-Aldrich Corp., St. Louis, MO, USA). Cells were maintained at 37°C in a humidified 5% CO2 incubator. For Eupatilin (Sigma, USA) treatment, cells were treated with different concentrations of Eupatilin (5, 10, and 20 mM) for 24 h.
Eupatilin
Manufactured by Merck Group
Sourced in United States
Eupatilin is a laboratory equipment product manufactured by Merck Group. It is a chemical compound used in various research and analytical applications. The core function of Eupatilin is to serve as a standard or reference material for analytical procedures. It can be utilized in techniques such as spectroscopy, chromatography, and other analytical methods to ensure the accuracy and reliability of experimental results.
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Lab products found in correlation
Lipofectamin 3000, by Thermo Fisher Scientific (2 mentions)
Fatal bovine serum, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Donkey anti rabbit and anti mouse immunoglobulin secondary antibodies, by Enzo Life Sciences (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Mk2206, by Merck Group (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Cell Signaling Technology (1 mentions)
Bay11 7082, by Merck Group (1 mentions)
5 and 6 carboxy 2 7 dichlorodihydro fluorescein diacetate h2dcfda, by Merck Group (1 mentions)
Collagenase a, by Merck Group (1 mentions)
Enzyme linked immunosorbent assay reader, by Agilent Technologies (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Sodium lauryl sulfate, by Merck Group (1 mentions)
Copovidone kollidon va64, by BASF (1 mentions)
9 protocols using eupatilin
1
Human Renal Cancer Cells Eupatilin Treatment
2
Eupatilin and PMA Modulate MUC5AC Expression
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Eupatilin and PMA were obtained from Sigma (St. Louis, MO, USA). RPMI-1640 medium, fatal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). TRIzol reagent was obtained from Life Technologies (Carlsbad, CA, USA). For cDNA synthesis, SuperScript II Reverse Transcriptase was purchased from Thermo Scientific (Thermo Fisher Scientific, Vilnius, Lithuania). A monoclonal b-actin anti-body was obtained from Sigma. Antibodies for phospho-ERK-1/2 (p-ERK), total ERK-1/2 (ERK), phospho-p38 (p-p38), total p38 (p38), phospho-JNK (p-JNK), and total JNK (JNK) were obtained from Cell Signaling Technology Inc. (Beverly, MA, USA). Anti-MUC5AC antibody (SC-21701 AF488) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Donkey anti-Rabbit and anti-Mouse immunoglobulin secondary antibodies were obtained from Enzo Life Sciences (Farmingdale, NY, USA). The study protocol was approved by the institutional review board of Wonkwang University Hospital (IRB No. 2019-01-028-001). Informed consent was confirmed by the IRB.
3
Evaluating Particulate Matter Signaling
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Fine atmospheric particulate matter NIST 2786, eupatilin, 5-(and 6)-carboxy-2′,7′-dichlorodihydro-fluorescein diacetate (H2DCFDA), and Collagenase A were obtained from Sigma-Aldrich (St Louis, MO, USA). Antibodies against p38 mitogen-activated protein kinase (MAPK, #9212), phosphorylated p38 MAPK (#4511), Protein Kinase B (Akt, #9272), phosphorylated Akt (#9275), Nuclear factor kappa B (NF-κB) p65 (#8242), phosphorylated p65 (#3036), and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH, #5174) were provided by Cell Signaling Technology (Danvers, MA, USA). An inhibitor of Akt (MK-2206) and an inhibitor of NF-κB p65 (BAY11-7082) were purchased from Sigma-Aldrich (St Louis, MO, USA). FPM was suspended in phosphate-buffered saline (PBS). eupatilin and inhibitors were dissolved in dimethylsulfoxide (DMSO). DMSO alone had no observable effect at concentrations used (data not shown).
4
Eupatilin's Effects on Glioblastoma Cells
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The LN229 and U87MG cells were seeded in 96-well culture plates at a density of 5×104 cells/well. Following 24 h, they were treated with 12.5, 25 or 50 µM eupatilin (Sigma-Aldrich) for 24, 48, 72 or 96 h. Control group cells were treated with 0.1% dimethylsulfoxide (DMSO; Sigma-Aldrich) in culture medium. Subsequent to treatment application, all cells were incubated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT; Sigma-Aldrich) solution for 24 h at 37°C. Then DMSO was added in each well and shaken for 10 min at room temperature. The optical density was measured with an enzyme-linked immunosorbent assay reader (BioTek Instruments, Inc., Winooski, VT, USA) at a wavelength of 570 nm. Each experiment was performed at least three times.
5
Protective Effects of Eupatilin on Cisplatin-Induced Ototoxicity
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The House Ear Institute-Organ of Corti 1 (HEI-OC1) cell line is a widely used auditory hair cell line derived from the cochlea of the immortomouse [28 (link)], and it was cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM; AG29643096; Hyclone) supplemented with 10% fetal bovine serum (FBS; 10099-141; Gibco) and grown under 33°C and 10% CO2. The cells were trypsinized at 80% confluence using 0.25% trypsin/EDTA (25200056; Life Technologies). eupatilin was purchased from MCE (HY-N0783; MCE) and is dissolved at a stock concentration of 50 mM in dimethyl sulfoxide (DMSO; SHBM2270; Sigma-Aldrich) and further diluted to the desired concentrations. HEI-OC1 cells were pretreated with varying concentrations of eupatilin for 2 h and then cotreated with the optimal concentration (30 μM) of cisplatin for 24 h (479306; Sigma-Aldrich), which is dissolved at a stock concentration of 1 mM in H2O.
6
Formulation Development of Eupatilin Tablets
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Standardized ethanol extract of A. asiatica and commercialized IR tablet dosage forms of DA-9601 were provided by Dong-A Pharmaceutical Co. Ltd. Eupatilin (purity over 99% w/w) hydrochloride and sodium lauryl sulfate were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Copovidone (Kollidon VA64) and poloxamer P407 were kindly provided by BASF Co. Ltd. (Ludwigshafen, Germany). Hydroxypropyl methylcellulose (HPMC K4M) was obtained from Shin-Etsu Chemical Co. (Tokyo, Japan). Microcrystalline cellulose (Avicel® PH 101) was obtained from Asahi Kasei (Tokyo, Japan). All other chemicals used were of reagent grade and were used without further purification.
7
Antioxidant and Chemical Profiling
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Trolox®, α-tocopherol, butylated hydroxytoluene (BHT), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH), fluorescein, sodium persulfate, sodium acetate, n-tetradecane (≥99%), quercetin-3-O-glucoside (≥90%), rosmarinic acid (≥97%), naringenin (≥95%), apigenin (≥95%), eupatilin (≥98%), ursolic acid (≥90%), rutin (≥94%), (E)-β-caryophyllene (≥98.5%), α-humulene (≥96%), (2E, 6Z)-farnesol (≥96%), benzyl benzoate (≥99%), and benzyl salicylate (≥99%) were acquired from Sigma-Aldrich (St. Louis, MO, USA). Dipotassium phosphate was obtained from J.T. Baker (Phillipsburg, NJ, USA). Luteolin-7-O-glucoside (≥98%), luteolin-7-O-glucuronide (≥98%), cirsimaritin (≥98%), acacetin (≥98%), scopoletin (≥98%), and salvigenin (≥98%) were acquired from ChemFaces (Wuhan, China). Vitexin (≥95%) was obtained from Phytolab (Vestenbergsgreuth, Germany). Luteolin (≥98%) was purchased from Chemcruz (Santa Cruz Biotech., Dallas, TX, USA). The mixture of n-alkanes C8-C25 was obtained from AccuStandard, Inc. (New Haven, CT, USA). Helium, air, hydrogen, and nitrogen (99.995%) for GC analysis were purchased from Messer (Bucaramanga, Colombia). Ethanol (96%) was acquired from Suquin S.A.S. (Bucaramanga, Colombia). Type I water (18.2 MΩ cm) was produced using a Millipore Direct-QTM (Merck, Darmstadt, Germany) purification system.
8
Ciliogenesis Induction in Confluent Cells
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Ciliogenesis in confluent-grown MEFs and NIH3T3 cells was induced by serum starvation (0.5% FCS) for at least 24 h. For DNA transfection, Lipofectamin 3000 (Invitrogen) was used following the manufacturer’s guidelines. Appropriate empty vectors were used as transfection control (TF-Ctrl). NIH3T3 cells were treated with 20 μM eupatilin (#SML1689; Sigma-Aldrich) or DMSO as a solvent control for 24 h.
9
Ciliogenesis Induction in MEFs and NIH3T3
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Ciliogenesis in confluent grown MEFs and NIH3T3 cells was induced by serum-starvation (0.5% FCS) for at least 24 hours. For DNA transfection, Lipofectamin 3000 (Invitrogen) was used following the manufactures guidelines. NIH3T3 cells were treated with 20 μM eupatilin (#SML1689; Sigma-Aldrich) or DMSO as a solvent control for 24 h.
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