Bioactive Substances in Safflower Flowers and Their Applicability in Medicine and Health-Promoting Foods

In vitro: HSAEC1-KT HSAECs (human small airway epithelial cells)	ELISA kits/concentrations of inflammatory cytokines (IL-6, IL-1β, and TNF-α) Western blot analysis/protein expression Calcium-sensitive fluorescent probe Fluo3/AM/intracellular calcium ion concentration Dual-luciferase reporter assay/transcriptional activities of NF-κB and AP1	9, 27, and 81 mmol/l
[74]
In vivo: rats In vitro: H9c2 (rat cardiomyoblast cells)	Double-staining method (with Tcc and Evans blue stains)/infarct size ELISA kits/cTnI, IL-6, and LdH levels Flow cytometry analysis, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling)/apoptosis level ELISA/caspase-3 activity CCK-8/cell viability Western blot analysis/protein expression Span diagnostic reagent kit/serum glutamic oxaloacetic transaminase Span diagnostic reagent kit/serum alkaline phosphatase Agappe diagnostic kit/serum total bilirubin	5 mg/kg HSYA; intraperitoneally injected 20 μM	[47]
In vivo: mice	SP kit (MMP-2, MMP-9, and COX- 2)/immunohistochemical analysis	1.125 mg/kg, 2.25 mg/kg, intraperitoneally, for 14 days
In vitro: H22 (murine hepatoma cells), HepG2 (human hepatocellular carcinoma cells)	Growth inhibition assay—CCK-8/rate of growth inhibition (IC50 80 μM for 72 h) Western blot analysis/protein expression Clonogenic assay/optical density at 570 nm	40, 60, 80, 120, and 160 μM, for 24, 48, and 72 h	[91]

Table 3: Continued.

Test organism	Type of test/parameter Concentration/dose		Authors
	Wound healing assay/cell migration, via Image-Pro Plus
In vitro: KYSE-30 cells (esophageal cancer cells)	CCK-8 assay/proliferative activity (optical density of cells) Microscopic measurement of the number of cells/cell invasion and cell migration assays Flow cytometric analysis of cell apoptosis/apoptotic rate of cells Western blot analysis/protein expression Serum levels of AFP, ALT, TBIL, and ALB/automatic analyzer	0,1, 1, 10, 20, and 50 μM, for 24, 48, and 72 h 20 μM, for 24 h	[92]
In vitro: A2780/DDP (ovarian carcinoma cells)	Cytotoxicity assay (CCK-8)/cell survival rates RTCA test (real-time cellular analysis)/monitoring cell proliferation Flow cytometry (Guava EasyCyte Plus)/evaluation of apoptosis	1 mg/ml 1 mg/ml, for 24	[45]
In vivo: mice	Electronic digital caliper/weekly measurement of tumor size and volume Western blot analysis/protein expression	1.1 g/kg body weight; once every three days, for five weeks