Kaempferol (Robigenin) is a natural flavonoid and an inverse agonist of ERRα and ERRγ. Kaempferol has a wide range of antitumor, anti-inflammatory, antioxidant, antibacterial and antiviral activities.

METHODS: Human breast cancer cells MDA-MB-231 were treated with Kaempferol (0.01-100 μM) for 72 h, and cell viability was detected by MST assay.
RESULTS: Kaempferol inhibited cell proliferation in a dose-dependent manner with an IC50 value of 43 μmol/L. [1]
METHODS: Human gastric cancer cells AGS and SNU638 were treated with Kaempferol (50 μM) for 8-24 h. The expression levels of target proteins were detected by Western Blot.
RESULTS: Kaempferol increased cleaved caspase-3 and -9 and decreased Bcl-2 levels in a time-dependent manner. [2]

METHODS: To investigate the effects on hemorrhagic shock mice, Kaempferol (10 mg/kg) was administered intraperitoneally to C57/BL6 mice experiencing hemorrhagic shock.
RESULTS: Pre-treatment of hemorrhagic shock mice with Kaempferol significantly reduced plasma TNF-α and IL-6 levels; restored MPO, SOD and MDA in the heart, lungs and liver; and increased HO-1 expression in the same organs. [3]
METHODS: To study the anti-neuroinflammatory effects, Kaempferol (25-100 mg/kg) was administered by gavage to BALB/c mice once a day for seven days, and LPS (5 mg/kg) was injected on the seventh day.
RESULTS: Kaempferol may be a promising neuroprotective agent to alleviate inflammatory responses and blood-brain barrier dysfunction by inhibiting HMGB1 release and downregulating the TLR4/MyD88 inflammatory pathway. [4]

Right atria or sinus nodal cells are homogenized in lysis buffer consisting of (50 mM Tris-HCl pH 7.5, 100 mM KCl, 1 mM ethylenediamine tetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 1 mM dithiothreitol, 0.1 mM phenylmethylsulfonyl fluoride, 0.5 mM Benzamidine, 20 mg/L Leupeptin, 20 mM sodium pyrophosphate, 50 mM NaF, and 50 mM sodium β-glycerophosphate), and total protein content is determined by the Bradford assay. Caspase-3 activity is determined by EnzChek Caspase-3 Assay Kit[3].

Kaempferol is prepared in DMSO (100 mM) and stored (-20°C), and then diluted with appropriate medium[2]. Ovarian cancer cells are seeded in 96-well plates at 2000 cells/well and incubated overnight before treatment with 0-160 μM Kaempferol for 24 hours in triplicates. The medium is removed, and the plates are freeze-thawed to lyse cells. Each well is added with 200 μL 1× CyQUANT cell lysis buffer containing 5x SYBR Green I and incubated at room temperature (RT) for 5 minutes. The reaction (50 μL) is transferred to PCR strip tubes and the fluorescent signal is measured at 90°C with a real-time Chromo4 PCR instrument. To ensure that cell proliferation assays are performed within a linear range of cell numbers, a standard curve is generated by seeding different amount of OVCAR-3 cells (based on counting with a hemacytometer) in a 96-well plate, and measuring genomic DNA abundance after overnight incubation. Three independent experiments are performed and data is pooled for statistical analysis[2].