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Black Cumin (Nigella sativa) and Its Active Constituent, Thymoquinone: An Overview on the Analgesic and Anti-inflammatory Effects


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For many centuries, seeds of Nigella sativa (black cumin), a dicotyledon of the Ranunculaceae family, have been used as a seasoning spice and food additive in the Middle East and Mediterranean areas. Traditionally, the plant is used for asthma, hypertension, diabetes, inflammation, cough, bronchitis, headache, eczema, fever, dizziness, and gastrointestinal disturbances. The literature regarding the biological activities of seeds of this plant is extensive, citing bronchodilative, anti-inflammatory, antinociceptive, antibacterial, hypotensive, hypolipidemic, cytotoxic, antidiabetic, and hepatoprotective effects. The active ingredients of N. sativa are mainly concentrated in the fixed or essential oil of seeds, which are responsible for most health benefits. This review will provide all updated reported activities of this plant with an emphasis on the antinociceptive and anti-inflammatory effects. Results of various studies have demonstrated that the oil, extracts, and their active ingredients, in particular, thymoquinone, possess antinociceptive and anti-inflammatory effects, supporting the common folk perception of N. Sativa as a potent analgesic and anti-inflammatory agent. Many protective properties are attributed to reproducible radical scavenging activity as well as an interaction with numerous molecular targets involved in inflammation, including proinflammatory enzymes and cytokines. However, there is a need for further investigations to find out the precise mechanisms responsible for the antinociceptive and anti-inflammatory effects of this plant and its active constituents. Georg Thieme Verlag KG Stuttgart · New York.
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The use of medicinal plants in various ailments
dates back to the earliest years of manʼsevolution
[1]. Nigella sativa L. (Ranunculaceae) is an indige-
nous herbaceous plant native to Southwest Asia
including Iran, India, and Pakistan. The plant
grows to a maximum height of about 4070 cm
and has finely divided foliage and pale blue and
white flowers. From the fruit capsules, many
small caraway-type black seeds are produced
(length: 2.5 to 3.5 mm and width: 1.5 to 2 mm).
In different languages the plant is known by vari-
ous names, e.g., black cumin, black seed, black-
caraway (English), Habbah Al-Sauda, seed of
blessing (Arabic), chernushka (Russian), çörek otu
(Turkish), and Cyah-daneh in Persian. For thou-
sands of years, the seeds of this plant have been
used as a spice and additive in bread, cookies,
and other dishes in many Asian and Eastern coun-
tries [2]. Therapeutic benefits of black cumin and
its active ingredients have been demonstrated in
many investigations [35].
Chemical Composition
N. sativa seeds contain various compositions in-
cluding moisture, oil, proteins (eight of the nine
essential amino acids), carbohydrates, vitamins,
and minerals [6, 7]. The percentage of ingredients
varies with the geographic distribution, time of
harvest, and cultivation methods [8].
In a study by Cheikh-Rouhou et al. comparing Tu-
nisian and Iranian varieties for their quality at-
tributes, the Tunisian variety contained 8.65,
28.48, 26.7, 4.86, and 40.0% of moisture, oil, pro-
teins, ash, and carbohydrates, respectively, while
analysis of the Iranian variety showed 4.08,
40.35, 22.6, 4.41, and 32.7% of the respective at-
tributes [9].
Black cumin seed is composed of fixed (stable)
and essential (volatile) oil responsible for many
For many centuries, seeds of Nigella sativa (black
cumin), a dicotyledon of the Ranunculaceae fam-
ily, have been used as a seasoning spice and food
additive in the Middle East and Mediterranean
areas. Traditionally, the plant is used for asthma,
hypertension, diabetes, inflammation, cough,
bronchitis, headache, eczema, fever, dizziness,
and gastrointestinal disturbances. The literature
regarding the biological activities of seeds of this
plant is extensive, citing bronchodilative, anti-in-
flammatory, antinociceptive, antibacterial, hypo-
tensive, hypolipidemic, cytotoxic, antidiabetic,
and hepatoprotective effects. The active ingre-
dients of N. sativa are mainly concentrated in the
fixed or essential oil of seeds, which are responsi-
ble for most health benefits. This review will pro-
vide all updated reported activities of this plant
with an emphasis on the antinociceptive and
anti-inflammatory effects. Results of various
studies have demonstrated that the oil, extracts,
and their active ingredients, in particular, thymo-
quinone, possess antinociceptive and anti-inflam-
matory effects, supporting the common folk per-
ception of N. Sativa as a potent analgesic and anti-
inflammatory agent. Many protective properties
are attributed to reproducible radical scavenging
activity as well as an interaction with numerous
molecular targets involved in inflammation, in-
cluding proinflammatory enzymes and cytokines.
However, there is a need for further investigations
to find out the precise mechanisms responsible
for the antinociceptive and anti-inflammatory ef-
fects of this plant and its active constituents.
Black Cumin (Nigella sativa) and Its Active Constituent,
Thymoquinone: An Overview on the Analgesic and
Anti-inflammatory Effects
Authors Bahareh Amin1, Hossein Hosseinzadeh 2
Affiliations 1Department of Pharmacology and Physiology, School of Medicine, Sabzevar University of Medical Sciences, Sabzevar, Iran
2Pharmaceutical Research Center, Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad
University of Medical Sciences, Mashhad, Iran
Key words
"Nigella sativa
"analgesia activity
"antiinflammatory effect
received Nov. 22, 2014
revised June 29, 2015
accepted July 16, 2015
Published online
Planta Med © Georg Thieme
Verlag KG Stuttgart · New York ·
ISSN 00320943
Prof. Dr. Hossein Hosseinzadeh
Pharmaceutical Research Center
School of Pharmacy
Mashhad University of Medical
Department of Pharmaco-
dynamics and Toxicology
136591775 Mashhad, I. R.
Phone: + 98 51 38 81 90 42
Fax:+ 985138823251
Amin B, Hosseinzadeh H. Black Cumin (NigellaPlanta Med
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beneficial effects attributed to N. sativa. Fixed oil contains appre-
ciable quantities of unsaturated fatty acids (linoleic, oleic, and in-
olenic acids) as well as saturated fatty acids in minor amounts
(arachidonic and eicosenoic acids). Dihomo-γ-lionoleic acid is a
powerful antioxidant, which exists in the fixed oil of seeds [3,
10]. Besides the fatty acid profile, it also consists of considerable
quantities of vitamin E (tocopherol α,β,andγ), retinol (vitamin
A), carotenoids (β-carotene), and thymoquinone (2-isopropyl-5-
methyl-1,4-benzoquinone). Fat-soluble vitamins comprise more
than 0.2% of the total oil content [11, 12].
Other ingredients of N. sativa include minerals such as potassi-
um, phosphorus, calcium, and iron, in greater quantities, as well
as zinc, magnesium, manganese, selenium, and copper in fewer
amounts. Alkaloids such as nigellimine, nigellidine, and nigelli-
cine are also present in trace amounts [1113]. Black cumin has
been known to contain considerable quantities of phytosterols
including β-sitosterol, avenasterol, stigmasterol, campesterol,
and lanosterol [1416].
Moreover, essential oil extracted from black cumin is of function-
al importance because of its rich volatiles, such as 18.424 % thy-
moquinone (TQ), 46% monoterpenes including p-cymene, α-
pinene, thymol (THY), dithymoquinone (DTQ, nigellone), and
thymohydroquinone (THQ) [1719]. The general chemical com-
position of N. sativa seeds has been presented in l
"Table 1 [6
8,1017]. Photodimerization of thymoquinone as a consequence
of exposure to sunlight during separation and extraction of the
quinones from the seed produces dithymoquinone. Among the
components isolated from the volatile oil of N. sativa, TQ has been
demonstrated to be the principal active ingredient [11, 20].
The chemical structure of main ingredients of N. sativa oil includ-
ing thymoquinone, dithymquinone, thymohydroquinone, p-
cymene, and thymol is shown in l
"Fig. 1.
Traditional Uses of Nigella sativa
Traditional uses of this amazing herb originate from the ancient
Egyptians, Greeks, and Romans. Black seed is referred to by the
Islamic prophet Mohammed as having healing powers for every
disease except death. N. sativa has been advocated by Ibn Sina
(Avicenna), the most famous physician and philosopher of the Is-
lamic world, as the bodyʼs energizing compound and as a remedy
Fig. 1 Chemical structure of the active ingredients
of oil of N. sativa L. seeds.
Table 1 The general chemical composition of N. sativa seeds [68, 1117].
Constituents Chemical composition % Range (w/w)
Oil Fixed oil Linoleic acid (Omega-6), Oleic acid, Palmitoleic acid Linolenic acid (Omega-3), Myristoleic acid, Diho-
molionolenic acid, Stearic acid, Eicosadienoic acid, Myristic acid, Arachidic acid, Behanic acid, Sterols
(β-sitosterol, avenasterol, stigmasterol, campesterol and lanosterol), Tocopherols (α,β,and γ) Thymo -
quinone, Retinol (vitamin A), Carotenoids (β-carotene)
Volatile oil Thymoquinone, p-Cymene, Carvacol, α-Pinene, β-Pinene, Longifolene, t-Anethole Thymol, Thymo-
hydroquinone, Dithymoquinone (nigellone)
Protein Glutamic acid, Arginine, Aspartic acid, Leucine, Glycine, Valine, Lysine, Threonine, Phenylalanine Iso-
leucine, Histidine, Methionine
Carbohydrate Glucose, Rhamnose, Xylose,Arabinose 24.940%
Minerals Calcium, Phosphorus, Iron, Potassium, Sodium, Zinc, Magnesium, Manganese, Copper, Selenium 3.77%
Saponins α-Hederin (melanthin), Hederagenin (melanthigenin) 0.013%
Alkaloids Nigelicine, Nigellimine, Nigellidine 0.01%
Other Vitamins Vitamin A, Thiamin, Riboflavin, Pyridoxine, Niacin, Folacin, Vitamin C 14%
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for fever, headaches, toothaches, and common colds. It was also
recommended as a soothing agent for skin disorders, wounds,
and external irritations [21]. According to the Holy Bible, black
seed is described as the melanthion by Hippocrates and Dioscor-
ides and as the Gith by Pliny [22].
In folklore medicine, the seeds and oil of N. sativa have been fre-
quently prescribed as a natural remedy for a diverse range of dis-
eases, such as, fever, cough, nasal congestion, bronchitis, asthma,
dyspnea, hypertension, diabetes, inflammation, milk production,
eczema, dizziness, and gastrointestinal disturbances. Its use in
pain conditions such as headaches, toothaches, and back pain
has also been recommended [23, 24].
Pharmacological Studies on the Biological Activities
N. sativa and its main active constituent TQ have been attributed
to numerous pharmacological activities [25]. Up to now, cyto-
toxic [26,27], antioxidant [2831], immune enhancement [32,
33], gastroprotective, hepatoprotective [34,35], antitussive [36],
hypolipidemic, and cardioprotective effects [3739], increased
milk production [40], hypoglycemic [41], hypotensive [42], and
antimicrobial [43,44] effects have been demonstrated. In addi-
tion, beneficial effects of N. Sativa and thymoquinone on convul-
sions [45, 46], depression [47], menʼs infertility [48], memory im-
provement [49], nociception, and inflammation [50, 51] have
been discussed.
Antinociceptive activity
Early work by Abdel-Fattah and coworkers demonstrated that
oral administration of N. sativa oil (50400 mg/kg) dose-depen-
dently attenuated the nociceptive responses caused by the acute
nociceptive stimuli such as the hot-plate test (thermal stimulus),
tail-pinch test (mechanical stimulus), and the early phase of the
formalin test (chemical stimulus). It also suppressed inflamma-
tory nociception induced by acetic acid writhing without affect-
ing spontaneous motor activity in mice. In that study, the system-
ic administration (2.510 mg/kg, p. o. and 16 mg/kg, i. p.) and
the intracerebroventricular (i.c. v.) injection (14 mg/kg) of thy-
moquinone to mice attenuated the response in not only the early
phase, but also the late phase of the formalin test. The authors
suggested that N. sativa oil and TQ produce antinociceptive ef-
fects through indirect activation of the supraspinal µ-1 and κ-
opioid, but not the δ-opioid receptor subtypes. However, none of
these receptor subtypes was implicated in the antinociceptive ef-
fect of TQ in the late phase of the formalin test [52].
In addition, Al Nagger et al. studied the neuropharmacological
activity of N. sativa extracts. Aqueous and methanolic extracts of
black cumin were found to elicit thermal and mechanical ant i-hy-
peralgesic effects in the hot-plate and Randall-Selitto tests, re-
spectively [53]. In another study, the aqueous extract of N. sativa
(500 mg/kg, p. o.) significantly increased the hot plate reaction
time in mice [54]. In an investigation by De Sousa and coworkers,
thymoquinone and its para-benzoquinone analogues showed a
significant reduction in the paw licking time of animals in two
phases of the formalin test [55]. In a recent randomized control
trial study on mice (30 animals in each group), the ethanolic ex-
tract of N. sativa showed antinociceptive effects against an acetic
acid-induced writhing test [56].
Neurotransmitters, such as gamma aminobutyric acid (GABA),
have an important role in descending inhibitory pathways of pain
[57, 58]. An increase in GABAergic tone has been demonstrated in
the anxiolytic and anticonvulsant activities of extracts as well as
TQ [45,59, 60].
Another potential antinociceptive effect of thymoquinone might
be through intervening in the serotonin/5-hydroxytriptamine (5-
HT) pathway. The role of 5-HT and norepinephrine via descend-
ing inhibitory pathways has also been demonstrated in the mod-
ulation of pain [61, 62]. Fixed oil of N. sativa (0.1 mL/kg/day) for
four weeks elicited antidepressant activity through the increased
brain 5-HT levels and decreased 5-HT turnover [47].
Anti-inflammatory activity
The volatile oil (0.66 ml and 1.55 mL/kg, i.p.) of N. sativa and thy-
moquinone (0.5, 1.0, 5 mg/kg, i. p.) exhibited a dose-dependent
anti-inflammatory effect against carrageenan-induced rat hind
paw edema and cotton seed pellet granuloma comparable to the
reference drug indomethacin (3 mg/kg, i. p.) [63]. Similarly, Al-
Ghamdi and coworkers demonstrated that the aqueous extract
of N. sativa possesses an anti-inflammatory action in carra-
geenan-induced paw edema similar to 100 mg/kg aspirin; how-
ever, it had no antipyretic activity on yeast-induced pyrexia [54].
In another study by Hajhashemi et al. as well as Ghannadi et al.,
both oral and intraperitoneal administration of polyphenols ex-
tracted from N. sativa as well as essential oil (200, 400, and
800 µL/kg) containing p-cymene (37.3%) and thymoquinone
(13.7%) suppressed the early and late phases of the formalin test,
acetic acid-induced writhing in mice, carrageenan-induced paw
edema, and croton oil-induced ear edema in rats. The authors re-
ported that treatment with naloxone failed to reverse the analge-
sic activities of both the polyphenols and essential oil [50, 64].
The methanolic extracts of different germination phases of
N. sativa showed significant anti-inflammatory and antinocicep-
tive effects in kaolin-induced rat paw edema and hot-plate tests,
respectively, throughout the duration of the study (1, 3, 6, and
18 h after the injection of kaolin). The highest effect was ob-
served from the 5th day to the 11th day of germination [65].
"Table 2 summarizes studies investigating the antinociceptive
and anti-inflammatory effects of N. sativa and its main active
constituent TQ in different experimental models.
Potential antioxidant and anti-inflammatory
mechanisms according to in vitro studies
Numerous molecular targets are involved in the antioxidant and
anti-inflammatory activities of N. sativa and its active agents. As
reported by Singh et al., essential oil and oleoresins obtained
from black cumin seeds showed antioxidant effects with chelat-
ing activity on ferrous ions, a scavenging effect on 1,1-diphenyl-
2-picrylhydrazyl (DPPH) radical, and activity for the linseed oil
system. Meanwhile, such effects by essential oils were higher
than that observed by synthetic antioxidants such as buthylhy-
droxytoluene (BHT) and buthylhydroxyanisole (BHA) [19]. Both
the crude fixed oil of N. sativa and TQ dose-dependently attenu-
ated thromboxane B2 as well as leukotrienes (LT) B4 and C4 in rat
peritoneal leukocytes stimulated with calcium ionophore
A23187. Consequently, they can inhibit the cyclooxygenase
(COX) and 5-lipoxygenase (5-LPO) pathways of arachidonate me-
tabolism, respectively [66]. The inhibitory effect of fixed oil on
leukotriene generation and lipid peroxidation was greater than
that of thymoquinone. It seems that other ingredients, such as
unsaturated fatty acids, may also contribute to these protective
effects. Both compounds also inhibited nonenzymatic peroxida-
tion of brain phospholipid liposomes. In this pathway, the po-
tency of TQ was about ten times more than that of oil. Similarly,
Amin B, Hosseinzadeh H. Black Cumin (NigellaPlanta Med
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Mansour et al. reported that TQ potently inhibits the formation of
leukotrienes in human blood cells by inhibiting both 5-LPO and
LTC4 synthase activity [67].
Marsik et al. reported that quinolones from N. sativa seeds inhib-
ited COX-1 and 2. The most active ingredient against COX-1 was
thymol, while most inhibitory effects on COX-2 were observed
with THQ and TQ. Meanwhile, THQ was more specific for COX-2
than TQ [68]. Production of free radical nitric oxide (NO) by the
inducible nitric oxide synthase (iNOS) enzyme was dose- and
time-dependently inhibited by TQ in the supernatants of LPS-
stimulated macrophages [69].
In another study by Vaillancourt et al., TQ significantly abolished
LPS-induced proinflammatory cytokines such as interleukin-
1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), metallopro-
teinase-13 (MMP-13), COX-2, and prostaglandin E2 in an in vitro
model of rheumatoid arthritis [70].
There are enough investigations demonstrating that reactive
oxygen species (ROS), including NO, which subsequently causes
oxidative stress and mediators such as eicosanoids, proinflam-
matory cytokines, and lytic enzymes released by the inf lamma-
tory cells macrophages, microglia, and neutrophils, play an im-
portant role in the induction and maintenance of acute or chronic
conditions of pain [7177].
The inhibition of these targets is therefore exploited to attenuate
chronic conditions of pain, especially neuropathic pain [78, 79].
Another potential neuroprotective effect of thymoquinone might
be through the activation of PPAR-γ(peroxisome proliferator-ac-
tivated receptor γ) [80]. The activation of PPAR-γreduces estab-
lished neuropathic pain in addition to preventing its develop-
ment [81].
In addition, the phosphorylation of p38 mitogen-activated pro-
tein kinases (MAPK), extracellular-regulated kinases, and tran-
scriptional factor, and nuclear factor-kappa B (NF-κB) induced
by LPS were also blocked by TQ. The activation of MAPK and nu-
clear NF-κB has been demonstrated to contribute to chronic pain
states such as neuropathic pain [8284].
Potential antioxidant and anti-inflammatory
mechanisms according to in vivo studies
In line with the in vitro anti-inflammatory effects of N. sativa, in
vivo studies confirm such effects. Oral administration of TQ and
its metabolite dihydrothymoquinone (25, 50, and 100 mg/kg for
5 days to mice) showed superoxide anion scavenger activity in
different tissues [85]. An anti-inflammatory effect of TQ has been
reported in experimental allergic encephalomyelitis (EAE) in an
animal model for human multiple sclerosis by increasing the re-
duced glutathione (GSH) in the spinal cord of animals [86].
Oral administration of TQ (80 mg/kg) to diabetic rats for 45 days
reversed the decreased activities of catalase (CAT), glutathione
peroxidase (GPx), and glutathione-S-transferase (GST), and in-
creased antioxidants such as GSH and vitamins C and E, while it
attenuated levels of lipid peroxidation markers such as malon-
dialdehyde (MDA) in the kidney and liver tissues of diabetic rats
[87]. Similarly, in a study on rabbits, glutamate reductase, GPx,
Table 2 Selected studies showing the different doses and routes of administration of N. sativa seed components tested in experimental models of nociception
and inflammation.
Compound Dose Route Effect Animal Ref.
Volatile oil 0.66 and 1.55 mL/kg i. p. Decreased carrageenan-induced hind paws edema Rat [63]
Decreased cotton seed pellet granuloma
Thymoquinone 0.5, 1.0, 5 mg/kg
Fixed oil 50400 mg/kg oral Increased animal reaction time to hot plate Mice [52]
Increased animal reaction time in tail-pinch test
Inhibited acetic acid-induced writhing
Inhibited pain response in the first phase of formalin test
Thymoquinone 2.510 mg/kg oral Increased animal reaction time to hot plate Mice [52]
Increased animal reaction time in tail-pinch test
Inhibited acetic acid-induced writhing
16 mg/kg i. p. Inhibited pain response in both first and second phases of
formalin test
14 µg/mouse i. c.v
Aqueous extract 500 mg/kg oral Increased animal reaction time to hot plate Rats [54]
Decreased carrageenan-induced paw edema
Aqueous and methanolic
1.25 g/kg i. p. Increased animal reaction time to hot plate Mice [53]
Increased animal reaction time to Randall-Selitto
Polyphenols 200, 400, and 800 µL/kg oral Inhibited the earlyand late phases of formalin-induced pain Mice [64]
200, 400, and 800 µL/kg Inhibited acetic acid-induced writhing
Inhibited carrageenan-induced paw edema
Essential oil 200, 400, and 800 µL/kg Inhibited croton oil-induced ear edema Rats [50]
Increased animal reaction time in light tail flick test
(only by essential oil)
Ethanolic extract 50 mg/kg i.p. Reduced acetic acid-induced writhing Mice [56]
Thymoquinone and para-
benzoquinones analogues
10 mg/kg i.p. Inhibited formalin-induced paw edema Mice [55]
Methanolic extract 1 g/kg oral Reduced kaolin-induced paw edema Rats [51]
Increased the reaction time on hot plate
Thymoquinone 2.5, 5 mg/kg i.p. Reduced neuropathic pain-induced allodynia and hyperalgesia Rats [109]
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and GST activity of the liver were induced by orally administered
TQ (10 and 20 mg/kg/day) for 8 weeks, which could explain the
effect of the black seeds in inhibiting the generation of bioactive
metabolites known to promote carcinogenesis and oxidative cell
damage. However, cytochrome enzymes (CYP1A2 and CYP3A4,
but not CYP2E1) were decreased by TQ treatment. Hence, it
seems that the potential drug interaction of TQ should be taken
into consideration [88]. Ulcerative colitis, a common clinical chal-
lenge, is a chronic inflammatory disorder in the gastrointestinal
tract with unknown etiology. In a gastric mucosal ischemia/re-
perfusion (I/R) injury model, thymoquinone (10 and 20mg/kg)
decreased gastric acid secretion, and acid output as well as the
gastric mucosal content/activity of lipid peroxide, the proton
pump, and myeloperoxidase (MPO) as a biomarker of inflamma-
tion along with the ulcer index. In contrast, GSH, total nitric ox-
ide, and superoxide dismutase (SOD) were decreased. Such ef-
fects were comparable to that of omeprazole as a reference drug
[89]. Moreover, thymoquinone showed protective effects on the
dextran sodium sulfate (DSS)-induced colitis with a significant
reduction in colonic MPO activity and MDA levels as well as an
increase in glutathione levels [90]. According to the work of Mah-
goub et al., acetic acid-induced colitis in rats was attenuated by
pretreatment with thymoquinone (10 mg/kg) for 3 days with a
comparable or even higher effect than sulfasalazine, a known
anti-colitis drug. In addition, the activity of MPO, platelet activat-
ing factor (PAF), and histamine, mediators of inflammation and
anaphylaxis, were decreased. In contrast, content of antioxidant
GSH was normalized [91].
However, in an earlier report by Juhás et al., TQ (0.05%) had no
protective effects against trinitobenzensulfonic acid (TNBS)-in-
duced colitis in mice [92]. In an ointment formulation, the black
seeds showed anti-inflammatory effects comparable to commer-
cial products [93].
The exposure of the brains of rats to lead revealed that TQ
(20 mg/kg, oral) for one month was able to ameliorate lead-in-
duced neuronal degeneration through inhibiting the microglial
reaction [94].
El-Mahmoudy and coworkers determined that the protective ef-
fects of TQ in streptozocine (STZ)-induced diabetic rats is mediat-
ed via inhibiting the phosphorylation of MAPK [69]. In an exper-
imental model of romatoid arthritis, the oral administration of
5 mg/kg/day thymoquinone significantly reduced the ser um lev-
els of proinflammatory cytokines, IL-1β, and TNF-αas well as
bone turnover markers such as alkaline phosphatase and tar-
trate-resistant acid phosphatase [70]. Similarly, thymoquinone
was found to inhibit IL-1β, 5-LPO, LTs B4 and C4, T helper 2 cyto-
kines (IL-4, IL-13, and IL-10), and eosinophils in the bronchoal-
veolar lavage fluid of ovalbumine-induced allergic asthma in
mice [95].
The aqueous extract of N. sativa (200 mg/kg, oral, for 5 days) and,
especially, the protein fraction of it possessed a potent protective
effect against tetrachloride carbon (CCL4)-induced hepatotoxic-
ity by a significant reduction of TNF-α, IL-1β, interferon gamma
(IFN-γ), and MDA levels, while it increased GSH content both in
serum and liver mice tissues. The ethanolic extract showed a less
protective effect in that study [96].
In one mouse model of experimentally induced morphine toler-
ance and dependence, brain oxidative stress and increased iNOS
expression were attenuated by coadministration with thymoqui-
none in mice. However, a morphine-induced progressive increase
in the brain glutamate level was not inhibited by this compound
[97]. Similarly, Gilhota et al. repor ted that thymoquinone (20 mg/
kg) significantly attenuated the immobilization-induced increase
in plasma NO levels of stressed mice [60]. Thymoquinone and
black cumin seed oil protected against the lipid peroxidation lev-
el after global cerebral ischemia-reperfusion injury in the hippo-
campus of rats [98]. In a recent work, N. sativa oil protection
against thioacetamide-induced liver injury was the result of scav-
enging the free radicals and protecting the liver cells against oxi-
dative damage [99].
In a stroke mode of rats, chloroform and petroleum ether ex-
tracts, and aqueous and hydroalcoholic extracts of N. sativa
(400 mg/kg, orally) for 7 days protected animals against focal ce-
rebral ischemia via a decrease in the concentration of thiobarbi-
turic acid reactive substance (TBARS) as well as via an increase in
the levels of GSH and antioxidant enzymes such as SOD and CAT
N. sativa and thymoquinone suppressed COX-2 and oxidative
stress markers by decreasing the levels of lipid peroxidation
MDA and increasing the level of SOD antioxidant enzyme in pan-
creatic tissue of STZ-induced diabetic rats [102].
Dariani and colleagues showed that the oral administration of
thymoquinone (10 mg/kg) attenuated seizure activity induced
by an intrahippocampal kainite model of temporal lobe epilepsy
in rats. Lipid peroxidation was attenuated by decreasing MDA, ni-
trite, and nitrate levels. Thymoquinone also lowered hippocam-
pal neuronal loss and mitigated astrogliosis [103].
In a model of vancomycin-induced nephrotoxicity in rats, thymo-
quinone (10 mg/kg, i. p.) for 8 days decreased kidney levels of
MDA; in contrast, it increased activities of SOD and GSHPx [104].
In addition, neuroprotective effects of thymoquinone have been
shown in the 6-hydroxydopamine-induced parkinsonism model
via decreasing the MDA and nit rite levels as well as increasing the
activity of SOD in the midbrain homogenate of rats [105].
Thymoquinone alleviated serum lipase, amylase, capase-1, and
MPO activity as well as proinflammatory cytokines (IL-1beta
and IL-18) in rats receiving ethanol and a high-fat diet. An opti-
mum protective effect was obtained with 100 mg/kg of thymo-
quinone [106].
Hepatic fibrosis was prevented by oral gavage of thymoquinone
(20 mg/kg or 40 mg/kg) through the regulation of phosphoryl-
ation of monophosphate-activated protein kinase (AMPK), liver
kinase B, and the inhibition of proinflammatory cytokines. The
expression of toll-like receptor 4 (TLR4) was also decreased by
thymoquinone [107]. Activation of this receptor has an important
role in inflammatory as well as neuropathic pain conditions.
After nerve injury, the activation of such a receptor induces mi-
croglia and astrocytes as well as the production of the proinflam-
matory cytokines in the spinal cord, leading to the development
and maintenance of inflammatory or neuropathic pain [108].
In a recent work, we examined the effects of thymoquinone on
the pain behavioral parameters in rats subjected to chronic con-
striction injury (CCI) of the left sciatic nerve as a neuropathic pain
model [109]. Anti-allodynia and anti-hyperalgesia effects ob-
served with thymoquinone (2.5 and 5 mg/kg, i. p.) were still evi-
dent after 2 weeks of treatment, which is in agreement with
those studies who suggest that the opioid system might have a
limited role in the antinociceptive effects of thymoquinone in
this model of neuropathic pain [52, 64]. We showed that antino-
ciceptive effects of thymoquinone might be attributed, in part, to
antioxidant activity, microglia activity inhibition, and a reduction
in the extent of apoptosis.
According to the Sethi et al. study, it is quite likely that many anti-
inflammatory activities assigned to N. sativa and its main constit-
Amin B, Hosseinzadeh H. Black Cumin (NigellaPlanta Med
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uent thymoquinone, including the effects on enzymes COX, iNOS,
and 5-LPO as well as proinflammatory cytokines such as TNF-α,
IL-1, and IL-6, may be mediated, in part, through the suppression
of NF-κB activation [110]. The expression of genes of many en-
zymes and cytokines are regulated by this transcription factor
[111]. The various molecular targets modulated by N. Sativa and
its major component thymoquinone are depicted in l
"Fig. 2 [69,
70,80, 95, 102,104, 105, 108].
Safety and Potential Toxicity
There are some studies suggesting that the therapeutic doses of
N. sativa and thymoquinone have low toxicity and a wide margin
of safety. Intraperitoneal administration of N. sativa (50 mg/kg)
for 5 days did not change the levels of enzymes and metabolites
in the liver and kidney of rats [23]. Zaoui et al. investigated the
acute and chronic toxicity of N. sativa fixed oil. Lethal dose 50%
(LD50) values obtained by single doses (acute study) were deter-
mined to be 28.8 mL/kg and 2.06 mL/kg after oral and intraperi-
toneal administration, respectively, in mice. In chronic toxicity,
rats were treated daily with an oral dose of 2 mL/kg for 12 weeks.
No changes were observed in the key hepatic enzymes levels, in-
cluding aspartate-aminotransferase, alanine-aminotranferase,
gamma-glutamyltransferase, and histopathological modifica-
tions in the heart, liver, kidneys, and pancreas after 12 weeks of
treatment. The serum cholesterol, triglyceride, and glucose levels
as well as the count of leukocytes and platelets decreased signifi-
cantly compared to the control animals. In contrast, hematocrit
and hemoglobin levels increased [112].
In another study, feeding Hibro broiler chicks with a diet contain-
ing 20 or 100 g/kg of N. sativa ground seeds for 7 weeks did not
adversely affect the animalsʼgrowth [113].
In an investigation using Sprague Dawley rats, the fixed oil of
black seed (4.0%) and the essential oil (0.30%) were safe, as sero-
logical factors such as liver and kidney functioning tests, serum
protein profile, cardiac enzymes as well as electrolyte balance re-
mained within normal values after 56 days. Similarly, indices of
red and white blood cells showed no significant variation. How-
ever, rats treated with the fixed oil moiety gained less weight
compared to the control, suggesting that black seeds could have
beneficial effects in obesity-related disorders [114].
In another study on mice, the LD50 of thymoquinone was
104.7 mg/kg and 870.9 mg/kg after oral and int raperitoneal ad-
ministrations, respectively, whereas the LD50 in rats was deter-
mined to be 57.5 mg/kg and 794.3 mg/kg after oral and intraper-
itoneal administration, respectively [115]. As reported by Man-
sour et al., thymoquinone was effective against CCl4-induced he-
patic damage only at a dose of 12.5 mg/kg, but not the higher
doses (25 and 50 mg/kg). It might be hypothesized that TQ at the
higher doses induces oxidative stress [116]. In a phase I clinical
study conducted on adult patients with advanced malignant can-
cers and treated with thymoquinone, oral doses of thymoqui-
none were tolerable for patients up to 2600 mg/day [117]. In a re-
cent study conducted by Tubesha and coworkeres on Sprague
Dawley rats, animals treated with 20 ml of thymoquinone-rich
fraction nanoemulsion (containing 44.5 mg/kg TQ) appeared
normal and there was no mortality or any signs of organ toxicity
during the 14-day experimental period [118]. Although, the oil of
N. sativa was marketed to treat disorders of skin such as acne and
eczema [119], two cases of allergic contact dermatitis were re-
ported after topical application of the oil in two persons who suf-
fered from maculopapular eczema [120,121].
Fig. 2 Different molecular targets of N. sativa and
thymoquinone in inflammatory diseases. CAT: cat-
alase; COX: cyclooxygenase; GPx: glutathione per-
oxidase; GSH: reduced glutathione; GST: gluta-
thione-S-transferase; IFN-γ: interferon gamma; IL:
interlukine; iNOS: inducible nitric oxide synthase; 5-
LPO: 5-lipooxygenase; p38MAPK: p38 mitogen-ac-
tivated protein kinases; MMP: matrix metallopro-
teinase; MPO: myeloperoxidase; NF-κB: nuclear fac-
tor-kappa B; PPAR-γ: peroxisome proliferator-acti-
vated receptor γ; SOD: superoxide dismutase; TLR-
4: toll-like receptor; TNF-α: tumor necrosis alpha
[69, 70, 80, 95, 102, 104, 105,108]. (Color figure
available online only.)
Amin B, Hosseinzadeh H. Black Cumin (NigellaPlanta Med
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N. sativa seeds contain a complex of more than 100 compounds,
some of which have not yet been studied or even identified. Un-
saturated fatty acids in fixed oil and essential oil components, es-
pecially thymoquinone, dithymoquinone, thymohydroquinone,
thymol, alkaloids, saponins, and vitamins as well as trace ele-
ments contribute to its health benefits associated with black cu-
min seeds. The original research articles published so far have
shown the antinociceptive and anti-inflammatory potential of
N. sativa seeds active ingredients, in particular, thymoquinone,
the main active constituent. Although the safety of N. sativa oil
and its active constituents has been investigated in some studies,
data on the bioavailability and other pharmacokinetic parame-
ters of this magic spice are still incomplete.
Furthermore, broad spectrum studies on specific cellular and
molecular mechanisms of action as well as controlled clinical tri-
als to prove its efficacy in humans are really needed to further as-
sess the application of N. sativa and/or thymoquinone as an anti-
nociceptive agent.
Conflict of Interest
The authors declare that there are no conflicts of interest.
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... CQ and HCQ have a temporary inhibitory effect on the entry of SARS-CoV-2 through changing the pH of the perfusion fluid. The issue can be reversed when either of the drugs is removed, while TQ increases endosomal pH, preventing SARS-CoV-2 entry into the cell, together with simultaneously attacking the virus due to the two single oxygens in the TQ molecule, thereby acting as both a shield and sword [81]. Moreover, the cationic amphiphilic nature of CQ and HCQ [82] results in their immobilization in the hydrophilic environment through the body, with the inability to reach the target organs. ...
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In today’s world, the outbreak of the coronavirus disease 2019 (COVID-19) has spread throughout the world, causing severe acute respiratory syndrome (SARS) and several associated complications in various organs (heart, liver, kidney, and gastrointestinal tract), as well as significant multiple organ dysfunction, shock, and even death. In order to overcome the serious complications associated with this pandemic virus and to prevent SARS-CoV-2 entry into the host cell, it is necessary to repurpose currently available drugs with a broad medicinal application as soon as they become available. There are several therapeutics under investigation for improving the overall prognosis of COVID-19 patients, but none of them has demonstrated clinical efficacy to date, which is disappointing. It is in this pattern that Nigella sativa seeds manifest their extensive therapeutic effects, which have been reported to be particularly effective in the treatment of skin diseases, jaundice, and gastrointestinal problems. One important component of these seeds is thymoquinone (TQ), which has a wide range of beneficial properties, including antioxidant and anti-inflammatory properties, as well as antibacterial and parasitic properties, in addition to anticarcinogenic, antiallergic, and antiviral properties. This comprehensive review discussed the possibility of an emerging natural drug with a wide range of medical applications; the use of TQ to overcome the complications of COVID-19 infection; and the challenges that are impeding the commercialization of this promising phytochemical compound. TQ is recommended as a highly effective weapon in the fight against the novel coronavirus because of its dual antiviral action, in addition to its capacity to lessen the possibility of SARS-CoV-2 penetration into cells. However, future clinical trials are required to confirm the role of TQ in overcoming the complications of COVID-19 infection.
... However, Also, vitamins (1-4%), proteins (25%), carbohydrates (33%), alkaloids (0.01%), fixed oil (22-38%), minerals (3.7-7%), essential oil (0.40-1.5%), and saponins (0.013%) are included in its composition. Different components in its content are responsible for the different effects of NS [34]. It can be revealed which compound is for which target with further studies. ...
Introduction: Acute necrotizing pancreatitis with a high mortality rate and the search for treatment continues. We investigated the protective effect of Nigella Sativa (NS), with antioxidant and anti-inflammatory effects, in experimental acute necrotizing pancreatitis. Materials and Methods: Thirty six male Wistar albino rats (weights 180-220 g) were randomised into four groups. Group 1 (Control): Rats were given standard mouse chow. No pro-drug or pancreatic intervention was performed. Group 2 (NS): In addition to their standard diet, rats were given 0.1 ml/100gr of NS orally for 3 days prior to the experiment. Group 3: Necrotizing pancreatitis was induced by retrograde administration of 3% Na-Taurocholate through the distal common bile duct of the rats on on experiment day. Group 4 (NS+ANP): Necrotizing pancreatitis was also formed in rats receiving 0.1ml/100 mg of NS for 3 days. Rats were given high-dose anesthesia 8 hours after the onset of pancreatitis. Immunohistochemical (TNF-a, MDA, MPO, Caspase), histological pancreatitis scoring and biochemical (LDH, Lipase, amylase) analyzes were performed from the blood and pancreatic tissue samples obtained. Results: There was no difference in histopathological, immunohistochemical and biochemical values between Group 1 and Group 2 (p>0.05). There were significiant differences between Group 4 and Group 3 in terms of histopathological, immunohistochemical and biochemical parameters (p<0.001). The pancreatitis findings of the Group 4 were found to be significantly milder than Group 3, which did not receive NS. Conclusion: NS pretreatment alleviates NaTaurocholate-induced experimental pancreatitis. NS firstly studied in experimental models of pancreatitis.
... Because of these ingredients, Nigella sativa can reduce cholesterol synthesis by hepatocytes and reduce its absorption from the small intestine. Nigella sativa also activates LDL receptors by reducing intracellular cholesterol leading to rapid clearance of LDL from the blood circulation [7][8][9]. ...
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Black cumin is one of the most important seed spice belongs to Ranunculaceae family, used in several cooked and processed products in India due to its antimicrobial and antioxidant properties. It is commonly used as a form of traditional medicine in Middle-Eastern countries due to having high antioxidant potential and immunity boosting properties. So, this study was planned to prepare the seed extracts by using different solvents and evaluating their antioxidant activity along with the determination of total flavonoids and phenolic compounds. The analysis revealed that the highest antioxidant activity (58.08%) of methanol seed extract was obtained and the highest level of total phenolic compounds was found in ethyl acetate extract as (148.59 mg/GAE/g). Moreover, the ethyl acetate extract showed the highest flavonoids content (870.0 mg/g QE) respectively. Thus, the findings showed that a high level of natural antioxidants can be derived from Nigella sativa seed extracts by methanol solvent.
In our study, it was aimed to examine the effects of thymoquinone, the active ingredient of Nigella sativa, which has known anticancer activities, and capsaicin, which is an important part of the endocannabinoid system, on the neuroblastoma cell line SH-SY5Y cells. SH-SY5Y cells were grown in culture in conventional culture flasks in DMEM medium at 37 °C and 5% CO2. When the cells were 70-80% confluent, morphological changes were examined under an inverted microscope. The cells were passaged into 96 microplates, and after passage, different concentrations of thymoquinone (2.5; 5; 10; 25; 50; 100; 200; 300 µM/ml) and capsaicin (0.675; 1.25; 2.5; 5; 10; 20; 50; 100 µM/ml) were applied to the cells. After administration, cytotoxic effect and proliferation rates/cell proliferation were analyzed by the MTT method. When compared to the control group, cultured cells treated with 200 and 300 μM thymoquinone and 5 and 100 μM capsaicin had reduced cell proliferation at statistically significant levels (p
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Neurodegenerative diseases, the leading cause of morbidity and disability is gaining increased attention as it imposes a considerable socioeconomic impact, due in part to the ageing community. Neuronal damage is a pathological hallmark of Alzheimer's and Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia and multiple sclerosis, although such damage is also observed following neurotropic viral infections, stroke, genetic white matter diseases and paraneoplastic disorders. Despite the different aetiologies e.g. infections, genetic mutations, trauma and protein aggregations, neuronal damage is frequently associated with chronic activation of an innate immune response in the CNS. The growing awareness that the immune system is inextricably involved in shaping the brain during development as well as mediating damage but also regeneration and repair, has stimulated therapeutic approaches to modulate the immune system in neurodegenerative diseases.
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Nigella sativa has attracted healers in ancient civilizations and researchers in recent times. Traditionally, it has been used in different forms to treat many diseases including asthma, hypertension, diabetes, inflammation, cough, bronchitis, headache, eczema, fever, dizziness and influenza. Experimentally, it has been demonstrated that N. sativa extracts and the main constituent of their volatile oil, thymoquinone, possess antioxidant, anti-inflammatory and hepatoprotective properties. In this review we aimed at summarizing the most recent investigations related to a few and most important effects of thymoquinone. It is concluded that thymoquinone has evidently proved its activity as hepatoprotective, anti-inflammatory, antioxidant, cytotoxic and anti-cancer chemical, with specific mechanisms of action, which provide support to consider this compound as an emerging drug. Further research is required to make thymoquinone a pharmaceutical preparation ready for clinical trials.
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Nigella sativa is an annual plant in the family Ranunculaceae, native to southern Europe, north Africa and southwest Asia. N. sativa seeds contain fixed oils, proteins, alkaloids (such as nigellicine and nigellidine), quinones (such as thymoquinone), saponin and essential oil. In traditional medicine, it has been used for several diseases such as antihelmintic, amenorrhea, asthma, diabetes, cough, diuretic and lactagogue. Pharmacological and biological activities of N. sativa and its constituents such as thymoquinone have been shown to be antioxidant, anti-ischemia, antinociceptive and anti-inflammatory, anticonvulsant and antitussive.
The essential oil of black cumin seeds, Nigella sativa L., was tested for a possible antioxidant activity. A rapid evaluation for antioxidants, using two TLC screening methods, showed that thymoquinone and the components carvacrol, t-anethole and 4-terpineol demonstrated respectable radical scavenging property. These four constituents and the essential oil possessed variable antioxidant activity when tested in the diphenylpicrylhydracyl assay for non-specific hydrogen atom or electron donating activity. They were also effective ·OH radical scavenging agents in the assay for non-enzymatic lipid peroxidation in liposomes and the deoxyribose degradation assay.
The present study was conducted to study the ameliorating effect of Nigella sativa seed oil (5ml/ kg body weight, 10 ml/kg body weight) on the liver damage caused by thioacetamide (20 mg/kg body weight) in albino rats for a period of eight week. A significant (p<0.05) improvement in the altered levels of bilirubin, albumin, total protein, alanine transaminase, alkaline phosphatase, y-glutamyl transferase was observed after treatment with 10 ml/kg body weight of Nigella sativa oil. Antioxidant enzymes like catalase, superoxide dismutase glutathione peroxidase, thiobarbituric acid reactive substances and reduced glutathione also showed significant improvement in their altered levels in Nigella sativa (10 ml/kg body weight) treated rats. The results confirm the ameliorating effect of Nigella sativa oil on liver injury caused by thioacetamide and suggest the ability of Nigella sativa oil in scavenging the free radicals and protecting the liver cell against oxidative damage. The histopathological examination of liver section also confirm the ability of Nigella sativa oil in decreasing the severity of histopathological injury caused by thioacetamide.
The anti-inflammatory activity of the volatile oil (v.o.) of Nigella sativa L. seeds and its active principle thymoquinone has been examined using carrageenan-induced oedema in rat hind paws and cotton seed pellet granuloma in rats. Both the v.o. and thymoquinone were found to produce a significant dose-dependent anti-inflammatory effect as evidenced by the significant inhibition of oedema formation and reduction of the granuloma weight. The v.o. (0.66 ml and 1.55 ml/Kg, i.p.) inhibited rat hind paw oedema formation by 64.12% and 96.26%, while thymoquinone (0.5, 1.0, 5 mg/Kg, i.p.) caused a reduction of 38.85%, 56.63% and 104.88%, respectively. Indomethacin (3 and 9 mg/Kg, i.p.) inhibited the oedema by 46.90% and 67.83%, respectively. In addition, the v.o. (0.33 ml and 0.66 ml/Kg, i.p.) inhibited granuloma formation by 17.64% and 46.86%, while thymoquinone (3 and 5 mg/Kg, i.p.) reduced granuloma weight by 13.04% and 48.09%. These effects were nearly comparable to indomethacin (3 mg/Kg, i.p.) which reduced granuloma weight by 34.37%. It was suggested that the anti-inflammatory activity of the v.o. of Nigella sativa seeds may be due to inhibiting the generation of eicosanoids and lipid peroxidation.
INTRODUCTION AND AIMS: Interleukin-1β (IL-1β) is an important inflammatory mediator of immune complex-induced glomerulonephritis (GN). As IL-1β is activated by Caspase 1 in an inflammasome-dependent intracellular process, we examined the functional role of the inflammasome components Nlrp3 and its adapter molecule Asc in autologous murine nephrotoxic serum nephritis (NTN), a model of immune complex-mediated GN. METHODS: NTN was induced in wild-type, Nlrp3- and Asc-deficient C57BL/6 mice after preimmunisation with rabbit IgG. After 21 days functional parameters (serum urea, total serum protein, serum cholesterol, albuminuria), renal histology and renal leukocyte infiltrates were compared between the three groups. Moreover, cellular and humoral immune responses against rabbit IgG were analysed. RESULTS: In comparison to wild-type, Nlrp3 knockout mice revealed significantly lower serum urea levels and developed a less pronounced nephrotic syndrome (less hypoproteinemia, hypercholesterolemia, and albuminuria). Consistently, renal leukocyte infiltrates were significantly decreased in Nlrp3-deficient mice compared to wild-type. The reductions in renal leukocyte counts were 45% for CD45+ leukocytes, 50% for CD4+ T-cells, 40% for CD8+ T-cells, 39% for CD11c+ dendritic cells, and 50% for F4/80+ mononuclear phagocytes. The reduced accumulation of renal leukocytes correlated with a significantly decreased expression of renal mRNA for inflammatory chemokines and cytokines like Ccl2, Ccl5, Cxcl10, Ccr6, Tnf, Inf-γ, and IL-1β. In contrast, NTN was not attenuated in Asc-deficient mice. Functional parameters and leukocyte infiltration were comparable between Asc knockout mice and wild-type controls. Analysis of systemic immune responses revealed a decreased cellular activation of Nlrp3-deficient splenocytes after re-stimulation with rabbit-IgG (reduced surface expression of CD69 on CD4+ T-cells, less secretion of Inf-γ). However, humoral immunity was exacerbated in Nlrp3-deficient mice, as indicated by increased autologous anti rabbit-IgG serum titers. Asc knockout mice also demonstrated an attenuated cellular immune response, with humoral immune responses being comparable to those of wild-type controls. CONCLUSIONS: In summary, these results identify Nlrp3 as an important pro-inflammatory mediator of immune complex glomerulonephritis. Surprisingly, we could demonstrate an inflammatory function of Nlrp3 independently of its adapter molecule Asc. Thus, therapeutic blockade of Nlrp3 receptors, but not Asc may be a new strategy in the treatment of immune complex nephritis.