Table 2. Pharmacological findings of biochanin-A validated via in vivo experiments.

(↑increase, ↓decrease, × inhibit, + activation).

Pharmacological application	Study model	Dose and route used	Findings	Reference
Diabetes	STZ-rat model	10 mg/kg bodyweight per os (p/o)	HbAIc level ↓ Glucose tolerance, Insulin resistance ↓	( Harini, Ezhumalai and Pugalendi, 2012)
		10–15 mg/kg p/o	Glucose digestion ↑ HbA1C levels ↓ Serum visfatin amount ↑	( Azizi, Goodarzi and Salemi, 2014)
		10–15 mg/kg p/o	FBS ↓ hyperglycemia-induced free radicals	( Sadri et al., 2017)
		15 mg/kg p/o	Nesfatin-1 ↑ Insulin ↑ FBG level ↓	( Eskandari Mehrabadi and Salemi, 2016)
		10–15 mg/kg p/o	Adiponectin ↑ insulin ↑ serum resistin ↓ serum adiponectin ↑	( Salemi et al., 2018)
		0.1, 1 and 5 mg/kg intraperitonial (i/p)	Mechanical allodynia ↓ Hyperalgesia ↓	( Chundi et al., 2016)
		10–15 mg/kg p/o	The concentrations of VEGF, TNF-α and IL-1β in retina ↓ blood sugar ↓ inflammation ↓ angiogenesis ↓	( Mehrabadi et al., 2018)
		10–20 mg/kg/day p/o	Diabetic nephropathy × Renal TGF-β expression↑	( Ahad, 2013)
	db/db diabetic mice model	10–50 mg/kg/day red clover extract (Red clover extract containing 9.6% biochanin-A) p/o	Hepatic PPARα/γ stimulation ↑ Hepatic fatty acid synthase levels ↓	( Qiu et al., 2012b)
	STZ-diabetic C57BL/6 mice	1 mg/kg/day p/o	Hepatic PPARα ↑ Lipid profile ↓ Glucose levels ↓	( Qiu et al., 2012a)
	High fat diet + streptozotocin	10, 20 and 40 mg/kg	Glucose tolerance ↓ Insulin resistance ↓ Insulin sensitivity, SIRT-1 expression ↑	( Oza and Kulkarni, 2018)
	Streptozotocin- Nicotinamide rat model	5 mg/kg	Fasting blood glucose ↓ Body weight ↑	( Ghadimi et al., 2019)
Dyslipidaemia	HFD Mice model	30mg/kg/day,60mg/kg/day, and 120mg/kg/day	LDL,total cholesterol ↓ Lipoprotein lipase and hepatic triglyceride lipase ↑	( Xue et al., 2017)
Obesity	HFD-induced obese rats.	p/o	HFD induced trace elements metabolism ↓ Glucose, insulin, ferritin, total cholesterol, phospholipids, free fatty acids ↓	( Antony Rathinasamy et al., 2020).
	C57BL/6 mice HFD	0.05% biochanin-A p/o	PPAR ↑ glucose 6-phosphatase and pyruvate kinase × Obesity-induced hepatic steatosis and insulin resistance ×	( Park et al., 2016)
Cardiovascular disorders	Transient coronary ligation in Sprague-Dawley rats	12.5, 25 and 50 mg/kg intragastrically	TLR4/NF-kB/NLRP3 signalling × AST, CK-MB, LDH enzyme releases ↓	( Bai et al., 2019)
	apoE-/- mice fed with Western diet.	50 mg/kg intragastrically	pro-inflammatory cytokines ↓	( Yu et al., 2020)
	isoproterenol-induced MI rats	10 mg/kg body weight subcutaneously	antioxidant levels ↑ lipid peroxidation and detoxifying enzyme systems ↓	Govindasami et al., 2020
Airway hyper responsiveness	Ovalbumin-Induced Airway Hyperresponsiveness BABL/c mice model	100 μmol/kg, p/o	Total inflammatory cells ↓ Eosinophils ↓ Neutrophils ↓ Th1-released IL-2 & TNF-α ↓ Th2-released IL-4 & IL-5 ↓	( Ko et al., 2011)
	Male Hartley guinea pigs	10–30 μm	baseline tension and cumulative OVA-induced contractions in isolated sensitized guinea pig tracheal × degranulation of mast cells× inflammation ↓	( Ko et al., 2011)
	PM2.5-induced lung toxicity in SD rats	5, 50,100 mg/kg intragastric administration	cell death ↓ release of pro-inflammatory mediators, MDA, LDH, AKP ↑ antioxidant enzymes levels ↑	( Xue et al., 2020).
Osteoarthritis	Osteoarthritis rabbit model	5–25 mM intra-articular injection	Cartilage damage ↓ MMP, NF-κB × TIMP-1 +	( D. Q. Wu et al., 2014)
Anti-microbial activity	BALB/c mice+ intragastric administration of Salmonella	2 μg/mL	Salmonella infection × AMPK/ULK1/mTOR pathway	Zhao et al., 2018a)
Neurological disorders:
Cerebral ischemia:	C57BL/6 male mice middle cerebral artery occlusion induced Focal cerebral ischemia	5,10 mg/kg intraperitoneal	GOT protein expression ↑ stroke lesion volume ↓	( Khanna, Stewart, Gnyawali, Harris, Balch, Spieldenner, Chandan K. Sen, et al., 2017)
	Ischemic stroke Rat model	10, 20, or 40 mg/kg/day	infarct size and brain edema ↓ SOD,GSH-Px ↑ MDA,NF-κB × Nrf2 nuclear translocation, HO-1 +	Guo et al., 2019)
	Middle cerebral artery occlusion (MCAO) rat model	10, 20, or 40 mg/kg/day	inflammatory response TNF-α and IL-1β levels, MPO activity↓ p38 signalling ↓	( Wang et al., 2015b)
Parkinson’s disease	SD Rats with Iron and Rotenone Co-treatment	30 mg/kg	malondialdehyde ↓ glutathione levels ↑ striatal dopamine depletion↓ behavioural impairments ↓ maintains redox equilibrium	( Yu et al., 2017)
	C57BL/6 with enhanced neonatal Iron and MPTP co-treatment	10–60 mg/kg	microglial p38 MAPK activation × behavioral and neurochemical deficits ↑	( Li et al., 2021b)
Alzheimer's disease	LPS-induced PD rats	12.5, 25, 50 mg/kg	inflammatory response ↓ MAPK signalling pathway × nitric oxide, IL-1, and TNF-α	( Wang et al., 2016).
	scopolamine-treated mice and natural aged cognitive deficit mice	40 mg/kg	AchE activity ↓ GSH ↑ thiobarbituric acid, ROS ↓ pyknotic neurons ↓	( Biradar et al., 2014)
	ovariectomized (OVX) rats	5,20,60 mg/kg	restore learning and memory deficits, MDA ↓ SOD and GSH-Px ↑	( Youn et al., 2016)