Background and the purpose of the study: Affinity-based target deconvolution is an emerging method for the identification of interactions between drugs/drug candidates and cellular proteins, and helps to predict potential activities and side effects of a given compound. In the present study, we hypothesized that a part of safranal pharmacological effects, one of the major constituent of Crocus sativus L., relies on its physical interaction with target proteins.Methods: Affinity chromatography solid support was prepared by covalent attachment of safranal to agarose beads.After passing tissue lysate through the column, safranal-bound proteins were isolated and separated on SDS-PAGE or two-dimensional gel electrophoresis. Proteins were identified using MALDI-TOF/TOF mass spectrometry and Mascot software.Results and major conclusion: Data showed that safranal physically binds to beta actin, cytochrome b-c1 complex sub-unit 1, trifunctional enzyme sub-unit beta and ATP synthase sub-unit alpha and beta. These interactions may explain part of safranal’s pharmacological effects. However, phenotypic and/or biological relevance of these interactions remains to be elucidated by future pharmacological studies.

Safranal (a monoterpene aldehyde) is the major volatile component of saffron which is responsible for the saffron unique odor. Several studies have shown the pharmacological activities of safranal including anti-oxidant, anti-inflammatory, cardioprotective, neuroprotective, nephroprotective, gastrointestinal protective, etc. This study was designed to review the pharmacological and medical effects of safranal and up-to-date previous knowledge. Moreover, some patents related to the pharmacological effects of safranal were gathered. Therefore, electronic databases including Web of Sciences, Scopus, and Pubmed for pharmacological effects and US patent, Patentscope, and Google Patent for patents were comprehensively searched by related English keywords from 2010 to June 2022. According to our review, most of the studies are related to the safranal effects on CNS such as antianxiety, analgesic, anticonvulsant, antiischemic, anti-tremor, memory enhancement and its protective effects on neurodegenerative disorders such as Alzheimer’, s, Parkinson and Huntington diseases. Other effects of safranal are antiasthmatic, antihypertensive, antiaging, anticataract, etc. Moreover, the protective effects of this agent on metabolic syndrome and diabetic nephropathy have been shown. Different mechanisms including anti-oxidant, anti-inflammatory, muscle relaxation, antiapoptotic, and regulatory effects on the genes and proteins expression related to signaling pathways of oxidative stress, inflammation, apoptosis, proliferation, etc. are involved in safranal pharmacological effects. Some patents for the prevention and/or treatment of different diseases such as liver cancer, sleep disorder, depression, cognitive disorder, obesity and PMS were also included. Based on the documents, safranal is considered a promising therapeutic agent although more clinical studies are needed to verify the beneficial effects of safranal in humans.

Background: Several biological and medical benefits of Saffron, Crocus sativus (Iridaceae), have been demonstrated. However, mechanisms of actions for purified constituents are greatly unknown. Objective: To examine the effects of Safranal, a main constituent of Saffron stigma, on cell viability and cytokine profile of peripheral blood mononuclear cells (PBMC) were examined. Methods: Effects of Safranal at 0. 1, 0. 5 and 1 mM concentrations were evaluated on cell viability and production of interleukin 4 (IL-4), IL-10 and interferon-γ (IFN-γ ) from non-stimulated and phytohemagglutinin (PHA) stimulated PBMCs, compared to 0. 1 mM dexamethasone and saline. Results: In stimulated cells, different concentrations of Safranal caused significant decrease of lymphocytes viability (p<0. 001 for all concentrations). All concentrations of Safranal inhibited IFN-γ and IL-10 secretion in stimulated cells (p<0. 01). In addition, high concentration of Safranal significantly decreased cell viability of non-stimulated PBMCs (p<0. 001). The effect of 1 mM Safranal on IL-4 secretion was less than dexamethasone (p<0. 05). Safranal showed a stimulatory effect on IFN-γ secretion in non-stimulated cells. The IFN-γ /IL-4 ratio at the presence of two higher Safranal concentrations both in non-stimulated and stimulated cells were significantly higher than those of control and PHA stimulated groups, respectively (p<0. 05). Conclusion: The IFN-γ /IL-4 ratio increases in the presence of Safranal which indicates an effect on Th1/Th2 balance. Therefore, Safranal may have therapeutic effects in inflammatory diseases associated with Th1/Th2 imbalance.

Gentamicin is an important aminoglycoside antibiotic. However its use is limited to serious and life threatening gram negative infections, because of its high nephrotoxicity potential in patients. There are reports that safranal, the active ingredient of saffron with antioxidant properties, exerts protective effect against ischemic injuries occurred by certain nephrotoxins including gentamicin. Therefore, in the present study, we examined the protective effect of safranal against gentamicin-induced nephrotoxicity in rat. After acclimatization, animals were randomly divided to three groups (8 rats /each group). On day one, each animal was placed separately in a metabolic cage for collecting 24-hour urine samples. On day two, after collecting  urine samples for measuring glucose and protein, the rats in group 1 received saline 1 ml/kg for 6 days, those in group 2 received gentamicin 80 mg/kg/day for 6 days, and the remaining rats in group 3 received safranal 0.5 ml/kg followed by gentamicin 80 mg/kg/day for 6 days. Injections were intraperitoneally. All the animals were euthanized 24 hours after the last dose. Blood samples were collected by cardiac puncture, and concentration of blood urea, creatinine, and urinary glucose and protein, as the indicators of nephrotoxicity were measured. Our results showed that in group 2, concentration of blood urea nitrogen p<0.01, creatinine p<0.05, urinary glucose p<0.001, and protein p<0.01 were significantly increased compared with the control and safranal-treated groups. There was no significant difference between the control and safranal-treated groups. Safranal exerts protective effects against gentamicin-induced nephrotoxicity in rat.

Background: Hexachlorobutadiene (HCBD) is a potent nephrotoxin in rodents, which can cause degeneration, necrosis and regeneration in renal tubular epithelial cells. It has been shown that safranal, the active ingredient of saffron, has a protective effect against ischemic injuries. The aim of this study was to examine the protective effect of safranal against HCBD-induced nephrotoxicity in rats.Method: Thirty Wistar albino rats were randomly divided in five groups. The rats received a single dose of corn oil 1ml/kg (group1), HCBD 50mg/kg (group 2), or safranal at doses of 0.5, 0.25 and 0.1 ml/kg one hour before HCBD (50mg/kg) injection (groups 3-5). All injections were carried out intraperitoneally. Urine samples were collected one day before, and one day after injections. On day 3 the animals were sacrificed and both kidneys were removed. The right kidney was fixed in formalin for histological examination and the left kidney was homogenized for measuring malondialdehyde (MDA). Blood samples were taken by cardiac puncture and used for the measurement of urea, creatinine, glucose and protein concentrations. Results: Blood urea concentration in HCBD treated group was significantly higher compared with group 3 (p<0.01) and groups 1 and 4 (p<0.001). There was no significant difference in urea concentrations between group 5 and HCBD treated group. Urinary concentration of glucose was significantly higher in group 2, compared with groups 1, 3 and 4 (p<0.001) No significant differences were observed in urinary glucose concentrations between HCBD- and safranal (0.1ml/kg)- treated groups. Concentration of protein was also significantly higher in group 5 than those of other tested groups (p<0.001). Conclusion: Safranal at doses of 0.25 and 0.5ml/kg has a protective effect against HCBD-induced nephrotoxicity in rats.