Table 1.

Effects of NS in pre-clinical studies of asthma.

Studies	Study material	Minimal active dose	Model	Negative control	Positive control	Effects								Notes & limitations
						Serum immunoglobulins	Inflammatory mediators	Inflammatory cells, BALF	Inflammatory cells, lung	Histamine release	Block H1 receptors	Relaxation of SM	Other
Chakravarty (1993)	Nigellone	11 µg/ml	Mixed peritoneal cells of egg albumin induced Wistar rats	N/A	N/A					↓				No control group (without Thymoquinone)
Gilani et al. (2001)	NS (70% aqueous-methanol extract)	0.1–3.0 mg/ml	Guinea pig trachea	N/A	N/A							+		No control group (without Thymoquinone)
Al-Majed et al. (2001)	Thymoquinone	50 µM	Guinea-pig trachea	N/A	N/A						+	+		No control group (without Thymoquinone)
Boskabady and Sheiravi (2002)	Aqueous extracts of NS	0.3 ml	Guinea pig trachea	Saline	Chlorpheniramine						+
Mansour and Tornhamre (2004)	Thymoquinone	3 and 10 µM	Human granulocytes	Untreated human granulocytes	N/A		↓ leukotrienes							No positive control
Abbas et al. (2005)	NS fixed oil	5 ml/kg/day injected (ip) for 17 days	Conalbumin sensitised (CD1) albino mice	Untreated mice	Dexamethasone	↓ IgG	↓ serum IL-2 & IL-12		↓				↓ blood eosinophil count
Büyüköztürk et al. (2005)	NS fixed oil	0.3 ml/day for 1 month	OVA sensitised BALB/c mice	Saline			No change in IL-4, IL-10 and IFN-γ in splenic mononuclear cells							No positive control
El Gazzar et al. (2006)	Thymoquinone + 10% DMSO	3 mg/kg TQ in 10%DMSO injected (ip) for 5 days	OVA sensitised BALB/c mice and lung cells	Saline + 10% DMSO	OVA + 10% DMSO	↓ OVA IgE & IgG1	↓ IL-4, IL-5, IL-13 ↑ IFN-γ in BALF	↓ eosinophils	↓ eosinophils				↓ goblet cells hyperplasia
Boskabady et al. (2008)	Methanol and dichloromethane extracts of NS	-0.8 g% of methanol extract-1.2 g% of dichloromethane extracts	Guinea pig trachea	Saline	Theophylline							+
Shahzad et al. (2009)	NS fixed oil	4 ml/kg/day injected (ip) for 7 days	OVA sensitised E3 rats	Saline	N/A	↓ Total IgE ↓ IgG1 ↓ OVA IgG1	↓ mRNA expression of IL-4, IL-5, IL-6 and TGF-β1 from lung cells ↓ nitric oxide in BALF	↓ eosinophils, macrophages & lymphocytes	↓ eosinophils, macrophages, lymphocytes				↓ bronchial and alveolar epithelial hyperplasia ↓goblet cells and collagen fibres	No positive control
Abd El Aziz et al. (2011)	Thymoquinone	3 mg/kg injected (ip) for 5 days in guinea pig-8 mg/kg injected (ip) for 21 days in rats	OVA sensitised guinea pig trachea Mast cells of egg-albumin sensitised rats	Saline	N/A					↓		+		No positive control
Balaha et al. (2012)	NS fixed oil	Oral NS oil 4 ml/kg/day for 31 days	OVA sensitised BALB/c mice	Saline	N/A	↓ Total IgE ↓ OVA IgE & IgG1	↑ BALF Th1 cytokines ↓ BALF Th2 cytokines	↓ leukocytes, macrophages & eosinophils					↓ Airway hyperresponsivness	No positive control
Saleh et al. (2012)	NS fixed oil	Oral NS oil 2.5 ml/kg/day for 3 weeks	OVA sensitised guinea pig Isolated rat peritoneal mast cells	Saline	N/A		↑ PGE2 in lung tissue ↓ leukotrienes			↓				No positive control
Keyhanmanesh et al. (2013)	Fractions of 20% methanolic extract of NS	(50, 100, 150, 200 mg/L)	Guinea pig trachea	Saline	Theophylline							+
Kalemci et al. (2013)	Thymoquinone	3 mg/kg/day injected (ip) for 5 days	OVA sensitized BALB/c mice	Saline	Dexamethasone								↓ Subepithelial and epithelial hyperplasia ↓ Number of mast and goblet cells
Keyhanmanesh et al. (2014)	Thymoquinone	0.3 mg/kg i.p.	OVA-sensitized guinea pig	Saline	N/A		↑ Blood IFN-γ	↓ Eosinophil ↓ Basophils					↓ Tracheal responsiveness ↓ Airway membrane hyperplasia ↓ Respiratory epithelial denudation ↓ Cellular infiltration ↓ Emphysema	No positive control
Saadat et al. (2015)	α-hederin	0.3 mg/kg i.p.	OVA-sensitized guinea pig	Saline	Thymoquinone			↓ Total WBC ↓ Eosinophils ↓ Basophils				+	↓ Tracheal contractile response to histamine
Fallahi et al. (2016)	α-hederin	0.2 mg/kg i.p.	OVA-sensitized Wistar rats	Saline	Thymoquionone		↓ IL-13 mRNA ↓ miRNA-126						↓ Pneumocyte and fibroblastic hypertrophy and hyperplasia ↓ Hyperemia ↓ Haemorrhage ↓ Edematous and exudative changes
Arabzadeh et al. (2016)	Ethanolic extract of NS	Oral 500 mg/kg/day for 3 weeks	Exercise-induced Wistar rats	Exercise-induced Wistar rats without NS	N/A								↓ thickness epithelial bronchi, tunica media (muscle) bronchi, and tunica adventitia bronchi ↓ number of goblet cells	No positive control

ip; intraperitoneal. OVA; ovalbumin. BALF; Bronchoalveolar lavage fluid. N/A; data not included in the original study.