TABLE 1.

Pharmacological activities of Daphnetin.

Pharmacological action	In-vivo/In -vitro study	Cell line/Animal	Method	Dose	Molecular mechanism	Effects/Targets	References
Anti-stress	In-vivo/in-vitro	Kunming mice Cortical neurons from SD rat brains	Unpredictable stressor	2 and 8 mg/kg	↓GRs	↓ in spatial learning and memory improves the cognitive deficits caused by chronic stress	Liao et al. (2013)
Neuroprotective	In-vivo	E18 C57BI/6 mice	NMDA induced excitotoxicity	20 and 40 mg/kg	× NR2B-containing NMDA receptors	× apoptosis × calcium overload	Yang et al. (2014)
Hepatoprotective	In-vivo In-vitro	male C57BL/6 mice	Oxidative stress induced hepatotoxicity	20, 40 and 80 mg/kg	↑ Keap1-Nrf2/ARE-Trx-1↓ASK1/JNK, P53 protein	↓ t-BHP in HepG2 cells ↑ Nrf2/Trx-1 ↑GSH, ↓ ROS	Lv et al. (2020)
Helicobacter Pylori infection	In-vitro	20 H. pyloristrains isloted from gastric antrum		6.25 or 12.5 μg/ML	↑ DNA damage, ↑recA ×membrane changes	↓ bab A, urel transcription and H. pylori adhesion to GES-1 cell line	Wang et al. (2019)
Lung protection	In-vitro/In-vivo	Mice	Endotoxin induced Lung injury	5, 10 mg/kg	× activation of macrophage and human alveolar epithelial cells, induced TNFAIP3 ↓pro-inflammatory cytokines	NF-Kb related signal pathway, anti-inflammatory potential	Yu et al. (2014b)
	In-vivo	C57BL/6 mice	L-arginine	2-4 mg/kg i.p	↓IL-6, TNF-α, MPO ↓JAK-2, STAT-3	↓infiltration and cytokine secretion in inflammatory cells	Yang et al. (2021c)
Rheumatoid arthritis	In-vivo	Rats	Freund’s complete adjuvant induced arthritis	2.25 and 4.5 mg/kg	↓IL-1, TNF-α and MIF	↓paw swelling and arthritic scores × inflammatory cells infiltration and articular cartilage degeneration	(Gao et al., 2008; Yao et al., 2011; Tu et al., 2012)
	In-vivo	Female Wistar rats	Collagen induced arthritis	1 and 4 mg/kg	↑ Foxp3	↓Th1/Th2/Th17	Yao et al. (2011)
					↓ activity of Th17 ↓ RORγt, NF-KB, CD77 ↓IL-10 ↑ Tregs	↓ paw swelling ×hyperplasia of synovial, destruction and degeneration of chondrocytes Modulate balance of Th17 and Tregs
Osteoporosis	In-vivo In-vitro	Sprague Dawley rats MC3T3-E1 pre osteoblasts	Dexamethasone induced	1 and 4 mg/kg	activate Wnt/GSK-3β/ß catenin signaling pathway	↓ body weight gain, bone mineral content and microstructure parameters	Wang et al. (2020b)
						↑ osteoblast proliferation, differentiation and mineralization
Hepatoprotective	In-vivo	Mice	Lipopolysaccharide/D-galactosamine induced liver failure	20, 40, 80 mg/kg	× Inos × COX-2 ↑ autophagy ↑ pro-autophagy protein expression	↓ ALT, AST ↓pro-inflammatory cytokines ↓MDA ↓myeloperoxidase ↑GSH, SOD level × MAPK, NF-kβ, NLRP3	Lv et al. (2018)
Liver cancer	In-vivo	Huh7 and SK-HEP-1		0, 5, 10, 50 and 100 µM	↑ G1 phase arrest	×cell viability	Liu et al. (2022)
						×tumorigenesis
						↑cell apoptosis
						×Wnt/ß catenin signaling
Nephroprotective	In-vivo	C57BL/6 mice	Cisplatin induced nephrotoxicity	2.5,5,10 mg/kg	↓ TNF- α, IL-1β, ROS, MDA	×NF-kB signaling pathway activate Nrf2 pathway	Zhang et al. (2018)
						↓ BUN, creatinine
						↓ renal injury
						↓inflammation, oxidative stress, apoptosis
	In-vivo	WT and Nrf2 mice	Cisplatin induced nephrotoxicity	20-40 mg/kg	↑SOD, GSH, SIRT1, SIRT6, HO-1, Nrf2, NQO1 ↓MDA, MPO	↓ BUN, creatinine ↓ renal injury ↓inflammation, oxidative stress, apoptosis	Fan et al. (2020)
Diabetic nephropathy	In-vivo	mesangial cells	High Glucose induced	0, 10, 20, 40 μM	↑ Nrf2 ×p-Akt ×p-p65	↓ ROS, MDA ↓TNF- α, IL-1β ↓IL-6, ↓fibronectin ↓collagen IV ↑ SOD activity ↓ cell proliferation	Xu et al. (2019)
Cerebral Ischemia/Reperfusion injury	In -vivo	C57BL/6mice	Middle cerebral artery occlusion	5, 10, 20 mg/kg	↓TNF- α, IL-1β, IL-6, TLR4	×TLR4/NF-kβ ↓IkBα degradation ↓neural cell apoptosis	Liu et al. (2016a)
	In-vitro	Hippocampal neuron	Reoxygenation induced lung injury	10, 20 and 40 µm	↑ Nrf2 ↑ HO1	× oxidative stress and neuronal apoptosis	Zhi et al. (2019)
Ischemic brain injury	In-vitro	HT22 cells	glutamate induced toxicity in hippocampal HT22 cell	5, 10, 25, 50, 75, and 100 μM/L	×NF-kB pathway	↑SOD, GSH ×TLR4/NF-kB pathway	Du et al. (2014)
Microglial activation	In-vivo	Murine microglia	Intracellular signal transduction	0–160 µm	×iNOS ×COX-2	↓ TNF-α, IL-1β, IL-6 NO, ×microglial activation ↓Th17 development ×NF-kB × MAPK × IKK /IkB PI-3K/Akt	Yu et al. (2014a)
Psoriasis	In-vitroIn-vitro	HaCaT human keratinocytes in Mice	imiquimod induced psoriasis like skin lesion	50-100 mg/ cm	↓ IL-1β, IL-6, IL-8, IL-17A, TNF- α, IL23A, MCP-1	pathway ×p65 phosphorylation ×nuclear translocation ↓erythema ↓scaling, epidermal hyperplasia, inflammatory cells infiltration	Gao et al. (2020)
Cell proliferation and Estrogenicity	In-vivo In-vitro	MCF-7 cells Female mice		17.5, 35, 70, 140 mg/kg	↓ Cyclin D1 ↑ p27	↑GO phase ↓G1 phase ↑S phase ↑G2 phase ↑M phase ↓cyclin/CDK2 ↓cyclin D1/CDK4 ×cyclin D1	Jiménez-Orozco et al. (2011)
Leukemia	In-vivo	Albino Wistar rats	Benzene induced	12.5, 25, 50 mg/kg	↑sphingosine1-phosphate receptor-1 ↓SGOT ↓Cytochrome P450 ↓CYP2E1	↓ NF-kB ↑Hematological parameter ↑nucleated bone marrow cells ↑megakaryocyte , SOD, GSH ↓MDA, ↓8- OhdG ↑albumin, total protein ↓BUN, bilirubin ↓prothrombin time	Pei et al. (2021)
Ovarian cancer	In-vitro	A2780		5, 10, 20, 40 μg/ml	↓ p-Akt ↓ p-mTOR ↑p-AMPK, LC3 II, p62	↑ROS production ×cell proliferation ↑apoptosis, autophagy, Blood count, Hemoglobin↓ proinflammatory cytokines	Fan et al. (2021a)
Human renal cell carcinoma	In-vitro	A-498 cells		10 and 50 µm	↑p38 MAP kinase ↑cytokeratin 8 and 18	MAPK Signaling pathway ×ERK1/ERK2 pathway ×S phase transition ×DNA synthesis	Finn et al. (2004)
Corneal inflammation and neovascularization	In-vivo	Male ICR mice	Alkali burn	(10-20 μmol/L) DAP eyedrops, q.i.d	↓HUVECs ↑STAT3, ERK, AKT	×corneal inflammation (↑VEGF-A) and neovascularization (↑TLR4/NLRP3)	Yang et al. (2022)
In various tumors	In-vivo	Female in bred BDF1 C57Bl/6	S180 sarcoma, MXT breast adenocarcino ma, C26 colon carcinoma	10,20 and 40 mg/kg	↑p38 MAP kinase ↑cytokeratin 8 and 18 ↑pro-apoptotic caspase-3	×mitogenic pathway ↓Cyclin D1 ×S phase ×Akt/ NF-kβ pathway ×proliferation	Jiménez-Orozco et al. (2020)
	In-vivo	Murine	Osteosarcoma LM8 cells	30-60 µm	↓ RhoA ↓Cdc42	↓intracellular stress fibers and filopodia	Fukuda et al. (2016)
Mitochondrial dysfunction and cell death	In-vitro	C57Bl/6 mice	Tert-butyl hydroperoxide	2.5,5,10 µg/ml	↑HO-1, SOD ↑NADPH, NQO1, GCLM ↑GCLC, BCl2 ↓ Bax, Caspase 3	×ROS production ×cytochrome c release, NLRP3 activation ↑Nrf2 pathway activate JNK and ERK	Lv et al. (2017)
CFA induced inflammatory pain	In-vivo	Murine	CFA	4 and 8 mg/kg	↓ spinal pro-inflammatory cytokines	×spinal glial activation × NF-kβ pathway ↑Nrf2 pathway/HO-1 signaling pathway	Yang et al. (2021a)
Inflammatory bowel disease	In-vivo	Mice	Fecal transplantation	16, 8, 4 mg/kg		↑ T reg cells development ↓ Th 17 cell differentiation	Ji et al. (2019)
Lipid metabolism	In-vitro	HepG2 cells		5, 20 and 50 µm	↑ PNPLA3	↓TG	Liu et al. (2019)
Insulin resistance	In-vitro	HepG2 cells		20 and 50 µm	↑ pAKT/AKT P13K	↑ glucose uptake	Liu et al. (2019)
Oxidative stress	In-vitro	HepG2 cells		5, 20, and 50 µm	↓ CYP2E1 and CYP4A ↑ Nrf2	↓ oxidative stress	Liu et al. (2019)
Angiogenesis	In-vivo	Rat	TNF and VEGF induced	9.375–900 µM	↓ c-Src, FAK, ERK1/2, Akt, VEGFR2, iNOS, MMP2	× angiogenesis ×migration ×invasion ×tube formation × NF-kβ pathway ×TNF-α induced IkBα degradation ×translocation of the NF- kβp65 protein ↑apoptosis	Kumar et al. (2016b)

Abbreviations: Inhibits; ↑: Upregulates, Increase; ↓: Downregulates, Decrease; CUS, chronic unpredictable stress; GRs: Glucocorticoid receptors; ALF, acute liver failure; APAP, acetaminophen; ASK1, Apoptosis signaling-regulating kinase 1; AREs, Antioxidant response elements; HO-1, Heme oxygenase-1; JNK, c-Jun N-terminal kinase; NF-κB, Nuclear factor-kappaB; Nrf2, Nuclear factor erythroid 2-related factor; 2NLRP3, Nucleotide-binding domain-like receptor protein 3 ROS, reactive oxygen species; Trx-1, Thioredoxin-1; Txnip, Thioredoxin-interacting protein; PALI, pancreatic acute lung injury; JAK-2, Janus kinase-2; STAT-3, Signal transducer and activator of transcription 3; VEGFR2, Vascular endothelial growth factor 2; iNOS, inducible nitric oxide synthase.