Iranian Journal of Basic Medical Sciences
Volume:25 Issue: 8, Aug 2022

Saffron, the stigmas of Crocus sativus L., has been mentioned extensively in the traditional reference texts as a herbal medicine. Many clinical trials have been conducted on this valuable herbal substance and its main constituents following numerous cellular and animal assessments. In the present review, we have collected almost all of these clinical studies to clarify how much knowledge has clinically been achieved in this field so far and which scientific gaps are needed to be filled by more studies. A comprehensive literature review was conducted through a two-round search. First, we performed a general search for identifying the human disorders against which saffron was studied. Then, we searched specifically for the combination of saffron keywords and each disease name. Scientific databases including Scopus, PubMed, and Web of science were used for this search. Studies were collected through electronic databases from their inception up to August 2021. The largest number of these clinical studies represent the investigations into saffron efficacy in different neurological and mental disorders, particularly depression. This substance has clinically revealed significant protective effects against various types of depression, age-related macular degeneration, and allergic asthma. In some cases, such as sexual dysfunction, cognitive and metabolic disorder, the effects of saffron are still clinically open to dispute, or there are limited data on its positive influences. Overall, saffron and its constituents have promising effects on human disorders; however, it needs more clinical evidence or meta-analyses to be confirmed.

Atherosclerosis is the thickening or hardening of the arteries which is caused by a buildup of atheromatous plaque in the inner lining of an artery. Hypercholesterolemia, inflammation, oxidative stress, and trimethylamine N-oxide (TMAO) are important risk factors for atherosclerosis. Therefore, this study aimed to review the anti-atherosclerotic effects of Lactobacilli and Bifidobacterium via improving lipid profile and reducing the effects of oxidative stress, inflammation, and TMAO. To prepare the present review, several databases such as Scopus, PubMed, and Google Scholar were searched, and relevant articles from 1990 until 2022 were selected and reviewed. The present review showed that Lactobacilli and Bifidobacterium reduce the risk of atherosclerosis in both in vitro and in vivo studies by breaking down or altering cholesterol metabolism with the help of their by-products and by reducing inflammation and oxidative stress and TMAO. Consumption of Lactobacilli and Bifidobacterium can be useful in prevention of atherosclerosis.

Neonatal necrotizing enterocolitis (NEC) is the most common gastrointestinal critical illness in neonatal infants. TRPM7 reportedly plays a role in human inflammatory bowel disease (IBD) and colorectal cancer, but the role of TRPM7 in the pathogenesis of NEC remains vague. The expression of TRPM7 was determined in intestinal tissues of NEC patients and lipopolysaccharide (LPS)-induced IEC-6 cells. Subsequently, a loss-of-function assay was performed to assess the effects of TRPM7 on cell apoptosis and inflammatory response in IEC-6 cells after LPS induction. Furthermore, the modulation of TRPM7 on TLR4/NF-κB and MEK/ERK signaling pathways was validated. The expression of TRPM7 was higher in the intestinal tissues of NEC patients compared with the normal human intestinal tissues. Moreover, the expression level of TRPM7 was elevated in LPS stimulation IEC-6 cells. Knockdown of TRPM7 enhanced cell viability and suppressed apoptosis, accompanied by the decreased Bax/Bcl-1 ratio and cleaved-caspase3 expression in LPS-induced IEC-6 cells. Additionally, TRPM7 silencing attenuated LPS-induced expressions and secretions of proinflammatory cytokines. Mechanistically, TRPM7 knockdown inhibited the TLR4/NF-κB activation, while enhancing the MEK/ERK activation in LPS-treated IEC-6 cells. Overexpression of TLR4 or inhibition of MEK attenuated the inhibitory effects of TRPM7 knockdown on LPS-induced apoptosis and inflammation in IEC-6 cells. Knockdown of TRPM7 attenuated LPS-induced IEC-6 cell apoptosis and inflammation by modulating TLR4/NF-κB and MEK/ERK pathways.

The effects of protein hydrolysates (FP) from Litopenaeus vannamei on oxidative stress, and autophagy gene expression was investigated in the NAFLD-induced rats.   For this purpose, twenty-four male rats were divided into four groups: Control, High-fat diet (HFD), FP20+HFD, and FP300+HFD (20 and 300 mg FP /kg rat body weight) and fed for 70 days.  The results indicated that the rat body and relative weight of the liver were not affected by experimental treatments (P>0.05) although the highest relative weight of the liver was observed in HFD treatment. The highest and lowest values for antioxidant enzymes and MDA concentration were observed in FP treatments (P<0.05). Also, the results showed that FP significantly decreased liver enzymes (ALT, AST) in the liver in comparison with HFD treatment (P<0.05). Plasma biochemical indices were investigated and the lowest amylase, ALP, fasting glucose, insulin, HOMA-IR, triglycerides, cholesterol, and inflammation cytokines (TNF-α, IL-6) were seen in the FP treatments which had a significant difference with HFD (P<0.05). Autophagy gene expression in the liver cells was affected by experimental diets and the lowest expression of Beclin-1 and Atg7 was observed in HFD and FP300 treatments. Interestingly, the highest expression of LC3-ɪ and P62 was seen in HFD and FP treatments, not in the control.  Overall, the results of this experiment indicated that FPs extracted from Whiteleg shrimp at 50 °C improve the oxidative status, glucose metabolism, and autophagy gene expression and could be used as a useful nutritional strategy in fatty liver prevention.

Atrial fibrillation (AF) is a common arrhythmia with atrial myocyte hypertrophy linked with stroke, heart failure, and increased mortality. Lysyl oxidase-like 2 (LOXL2) involves the cross-linking of collagen in the extracellular matrix (ECM). In the present study, we investigated the roles and underlying mechanisms of LOXL2 on cardiomyocyte hypertrophy.  The expression of LOXL2 mRNA and protein were detected in angiotensin II (Ang II) treated rat cardiomyocytes H9c2 by RT-qPCR and western blot. Small interfering RNA (siRNA) mediated LOXL2 gene silencing was used to evaluate cardiac hypertrophy and related markers. Also, the protein expression of EMT markers and Smad3/NF-κB pathway was determined by western blot.  Ang II significantly increased mRNA and protein expressions of LOXL2 and increased mRNA levels of myocardial hypertrophy markers, including ANP, BNP, and β-MHC in H9c2 cells. Silencing of LOXL2 significantly suppressed Ang II-induced hypertrophy and reversed the increase in ANP, BNP, and β-MHC mRNA levels. Also, EMT markers’ expressions, as evidenced by increased E-cadherin and decreased vimentin, α-smooth muscle actin (α-SMA), fibroblast-specific protein (FSP), and collagen 1A1. Mechanistically, we found that LOXL2 silencing suppressed protein expressions of TGF-β1, p-Smad3, and p-NF-κB in Ang II-stimulated H9c2 cells. LOXL2 silencing also attenuated Ang II-induced increased expression and content of proinflammatory cytokines IL-1β (H) and TNF-α.  Our data speculated that LOXL2 might be a potential contributing factor to Ang II-induced cardiac hypertrophy, and TGF-β1/Smad3/NF-κB is involved in a signal axis and might be a potential strategy in treating cardiac hypertrophy.

Photochemical internalization (PCI) is an important type of photodynamic therapy for delivering macromolecules into the cytosol by the endocytosis process. In this study, 6-mercapto-1-hexanol (MH) was used to functionalize the gold nanostructure as a primer for surface modification to improve conjugation of multi-agents such as protoporphyrin IX (Pp-IX) and folic acid with gold nanoparticles (PpIX/FA-MH-AuNP) to facilitate the photochemical internalization. After surface modification of AuNPs with MH, PpIX and FA are bonded to the surface of the MH-AuNPs through the coupling reaction to produce the desired conjugated AuNPs. In the next step, the synthesized nanostructures were characterized by different methods. Finally, after selecting specific concentrations, light treatments were applied and cell survival was measured based on MTT analysis. Also, in order to better study the morphology of the cells, they were stained by the Giemsa method. The SPSS 16 software was used for data analysis By surface modification of the nanostructure with MH and then conjugation of FA to it, the incubation time of the drug in PpIX/FA-MH-AuNP was reduced from 3 hr to 30 min. Also, at each light dose, cell death in the presence of PpIX/FA-MH-AuNP was significantly reduced compared with unconjugated conditions (P<0.001). Under these conditions, the ED50 for PpIX and PpIX-MH-AuNP and PpIX/FA-MH-AuNP at a concentration of 2.5 μg/ml is 8.9, 9.1, and 6.17 min, respectively. The results show that the PCI of PpIX/FA-MH-AuNP increases the selective phototoxicity efficiency on cancer cells compared with the conventional process of photodynamic therapy.

Sleep deprivation is a common health problem in modern society and is negatively associated with deleterious effects on cognitive functions such as learning and memory ability. This study was undertaken to provide a detailed account of the effect of chronic ozone intraperitoneal injection on the deleterious effects of sleep deprivation on brain function in rats.  Sleep deprivation was induced using the modified multiple platform model. The rats received REM sleep deprivation with an intraperitoneal injection of ozone or midazolam for 28 days. The effects of ozone on REM sleep deprivation-induced hippocampus-dependent learning and memory deficits were studied by the following approaches: Morris water maze (MWM) tests were used to evaluate spatial learning and memory of rats. Morphological changes in the brain were evaluated using hematoxylin and eosin (HE) staining. RNA-sequence was performed to seek a common mechanism. The expression of the targeted gene was examined by qPCR and Western blotting. Ozone intraperitoneal injection reversed spatial learning and memory deficits associated with REM sleep deprivation, ameliorating pathological brain damage and down-regulating the hippocampal expression of Sema3A in rats after REM sleep deprivation.  Ozone intraperitoneal injection alleviated sleep deprivation-induced spatial learning and memory deficits by protecting hippocampal neurons via down-regulation of the expression of Sema3A in the hippocampus.

Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic complication characterized by many side effects. Coenzyme Q10 (CoQ10) is a protective lipophilic molecule with an extensive range of biological functions, but its possible protective effect on the ovary in OHSS has not as yet been studied. The present study aimed to investigate the potential protective effects of CoQ10 on ovarian histological and molecular alterations in an experimental model of OHSS in rats.  Thirty female (2 months old) Wistar rats were randomly divided into 6 equal groups: control, OHSS, OHSS+CoQ10 (OHSS+ 200 mg/kg CoQ10 for 10 days), OHSS+ cabergoline (CAB) (OHSS+ 100 µg/kg CAB for 6 days), and CoQ10 and CAB (rats receiving similar doses to treatment groups.( In the end, the effects of treatments were assessed by measuring expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in the ovary via western blotting, ovarian histomorphological alterations assessments, and serum estradiol and progesterone levels via ELISA.  There were histological alterations in OHSS groups, including the elevation of diameter and numbers of the corpus luteum and atretic follicles, and decreasing follicular reserve count, hyperemia, and hemorrhage at ovarian stroma. Treatment of OHSS groups with CAB and CoQ10 could decrease histological changes, serum estrogen and progesterone, and overexpression of VEGF and COX-2 proteins. Our results showed that ovarian histological and molecular alterations observed in experimental OHSS can be ameliorated by administration of CoQ10, indicating that CoQ10 can be used as new supportive care for OHSS patients.

Rotavirus (RV) is one of the most significant pathogens associated with childhood diarrhoeal deaths worldwide. Elevated cytoplasmic calcium is required for RV replication, but the underlying mechanisms responsible for calcium influx remain poorly understood. The Calcium-sensing receptor (CaSR) is an important Ca2+ sensor that regulates the transport of Ca2+ into or out of the extracellular space by affecting the status of Ca2+ ion channels on the membrane of cells. Currently, the function of CaSR in RV replication is unclear.  We evaluated the mRNA and protein levels of CaSR in RV-infected cells using qRT-PCR and Western blotting, respectively. Furthermore, we silenced or overexpressed CaSR in Caco-2 cells using siRNA or a CaSR gene contained adenovirus (Adv-CaSR). qRT-PCR, plaque assay, and Western blotting were used to determine the synthesis of virus genomic RNA, production of progeny virion, and the levels of viral proteins. The content of Ca2+ in cells was observed under confocal microscopy.  Compared with control cells, the RV-infected cells presented significantly decreased CaSR expression. Moreover, adenoviral-mediated over-expression or induction of CaSR by R568 greatly inhibited the RV RNA synthesis, protein expression, and formation of viroplasm plaques, thereby suppressing RV replication. In contrast, CaSR-silenced cells exhibited significantly enhanced RV replication. Compared with the Adv-Control group, the concentration of cytosolic Ca2+ significantly decreased in the Adv-CaSR group.  These findings demonstrated that CaSR is a potential target for inhibition of RV replication. Therefore, enhancing the expression of CaSR might protect hosts from RV infections.

Previous studies have shown that ulinastatin (UTI) alleviates pulmonary edema in acute lung injury (ALI) caused by lipopolysaccharide (LPS), although the mechanism behind this action is uncertain. This research aimed to identify the fundamental mechanism by which UTI alleviates pulmonary edema. We established a model of acute lung injury (ALI) in rats by using LPS as the inciting agent.The control, LPS, and LPS+UTI groups were each comprised of a specific number of randomly selected Wistar rats. We evaluated lung injury and determined pulmonary edema. The concentrations of TNF-α, IL-1β and IL-6 in BALF and the expression levels of α1Na, k-ATPase, β1Na, K-AtPase, α-ENaC, β-ENaC, γ-ENaC, Zonula occludens (ZO)-1, Occludin, Caludin-5, PI3K, Akt, TLR4, MyD88 and NF-ƘBwere identified in lung tissues. The presence of UTI was associated with a reduction in lung pathological alterations, lung injury scores, the lung W/D ratio, and MPO activity, in addition to the improved gas exchange (P<0.01). Furthermore, UTI alleviated EB leakage and stimulated AFC (P<0.01). Importantly, UTI increased the expression of ZO-1, Occludin, Caludin-5, α1Na, K-ATPase, β1Na, K-AtPase, α-ENaC, β-ENaC, and γ-ENaC (P<0.01). Furthermore, UTI inhibited the inflammatory response, enhanced the expression of PI3K and Akt and hindered TLR4, MyD88, and NF-ƘB expression (P<0.01) in lung tissues. Our results demonstrated that UTI attenuated pulmonary edema by reducing pulmonary permeability and promoting AFC through inhibiting the inflammatory response, and the mechanism is related to promoting PI3K/Akt signaling pathways and suppressing TLR4/MyD88/NF-ƘB signaling pathways.

Dietary supplementation combined with exercise may potentiate the beneficial effects of exercise by reducing exercise-induced oxidative stress and improving mitochondrial quality and capacity. In this study, the effects of creatine monohydrate (CrM) supplementation with low and high-intensity exercise on mitochondrial biogenesis regulators, Nrf2 anti-oxidant signaling pathway and muscle damage levels were investigated.  Balb/c male mice were divided into six experimental groups: control, control+CrM, high-intensity exercise, high-intensity exercise+CrM, low-intensity exercise, and low-intensity exercise+CrM. Mice were given CrM supplementation and at the same time, low and high-intensity exercise was applied to the groups on the treadmill at 30min/5day/8week. Then, mitochondrial biogenesis marker (PGC-1α, NRF-1, TFAM), Nrf2 and HO-1 protein expressions, total oxidant-anti-oxidant status level, and histopathological changes were investigated in serum and muscle tissue.  Exercise intensity and CrM supplementation were found to be effective factors in mitochondrial biogenesis induction via the PGC-1α signaling pathway. Nrf2 and HO-1 protein levels increased with exercise intensity, and this result was directly related to serum oxidative stress markers. In addition, CrM supplementation was effective in reducing exercise-induced muscle damage.  This combination induced skeletal muscle adaptations, including mitochondrial biogenesis and enhanced anti-oxidant reserves. This synergistic effect of dietary supplementation with low-intensity exercise may be valuable as a complement to treatment, especially in diseases caused by mitochondrial dysfunction.

Multiple sclerosis (MS) is a chronic inflammatory disease affecting sensory and motor function in the central nervous system. Physical activities in the prevention and treatment of such conditions have shown promising results. However, their mechanisms of action have not been fully known yet and need further study. The present study aimed to evaluate the preventive effect of swimming exercise on some liver factors involved in inflammation and MS.  In this study, experimental autoimmune encephalomyelitis was induced in C57BL/6 mice, and the effect of a 6-week swimming exercise on the levels of fetuin-A, AMP-activated protein kinase (AMPK), and Nicotinamide adenine dinucleotide (NAD+) in their liver tissue was investigated by western blot analysis and NAD+ colorimetric assay. The study showed that EAE induction substantially (3.5 - fold) enhanced the fetuin-A levels and caused a reduction in AMPK and NAD+ amount. This is when doing 6 weeks of swimming exercise reduced fetuin-A to slightly above control. Also, levels of AMPK and NAD+ markedly increased in C57BL/6 mice with EAE. Doing regular exercise may limit the body’s inflammatory responses and reduce the severity of MS by regulating the expression of fetuin-A and increasing AMPK and NAD⁺ levels in liver tissue.

Neobavaisoflavone (NBIF) is an isoflavone isolated from Psoralea corylifolia L. It can effectively regulate the redox state as a natural anti-oxidant and show some anti-inflammatory activity. However, its molecular mechanism is poorly studied. In this study, RAW264.7 cells were treated with lipopolysaccharide (LPS) to investigate the anti-inflammatory activity and potential NBIF mechanism. RAW264.7 cells were treated with LPS (62.5 ng/ml) and exposed to different concentrations of NBIF (0.01, 0.1, and 1 μM) for 24 hr. Inflammatory cytokines of RAW264.7 cells were measured by the Griess method, ELISA, and western blot. Phagocytosis of RAW264.7 macrophages was measured by FITC-dextran uptake assay. The phosphorylation protein expression levels of MAPKs (JNK, p38, and ERK), NF-κB p65, IκBα, and IκB kinase were analyzed by western blot. The results were analyzed using one-way ANOVA with Tukey’s multiple comparison test. NBIF significantly inhibited NO and ROS production by down-regulation of iNOS and COX-2 protein expression. Additionally, the amount of release and protein levels of inflammation cytokines IL-6, IL-1β, and TNF-α were significantly decreased by NBIF. Moreover, FITC–dextran uptake assay by flow cytometry presented that NBIF significantly enhanced the phagocytic capacity of RAW264.7. Mechanistically, NBIF significantly down-regulated MAPK activation and inhibited the nuclear translocation of NF-κB p65.  The present study demonstrates that NBIF inhibited inflammation and enhanced the phagocytic capacity of RAW264.7 cell-related MAPKs and NF-κB signaling pathways induced by LPS. These findings suggest that NBIF may have clinical utility as an anti-inflammatory agent.

CB1 antagonism arbitrates a dormant shape to the endocannabinoid system that alleviates diverse pathological incidents of diabesity. The present study pursued the synthesis and evaluation of thiazolidine derivative (BAC) having pleiotropic action on CB1R, with or without AM251 (selective antagonist of the CB1 receptor) against high-fat diet (HFD) induced diabesity in C57BL/6 mice.  A molecular docking study for CB1 antagonistic potential was conducted by Maestro 11.4 program (Schrodinger Inc., USA), and the thiazolidine derivative BAC was synthesized. The assessment of varied parameters including anthropometric, neurobehavioral, hyperglycemia, dyslipidemia, oxidative stress, and inflammatory cytokines was evaluated in HFD-fed animals as compared with individual and combined treatments of BAC and AM251.  Incomparable to AM251, the treatment of BAC was reported for a significant reduction in food intake and obesity, diabetic biomarkers, lipid profile, oxidative stress, and proinflammatory cytokine release. Moreover, the BAC treatment showed no significant alteration in neurobehavioral activity, including anxiety and depression.  The preliminary in silico study suggests that BAC has a close interaction with CB1 antagonism but has no sign of neurobehavioral alteration. Simultaneously, this compound showed significant ability to ameliorate diversity by the underlying mechanisms of minimizing oxidative stress, regularizing the lipid profile, and reducing pro-inflammatory cytokines.

Ischemia-reperfusion injury is a life-threatening clinical problem that can occur after transplantation or a number of clinical procedures. The purpose of the study was to investigate the effects of Ambroxol on kidney damage caused by experimentally induced ischemia-reperfusion injury in rats.  Wistar albino rats were divided into 3 groups: Control (CTR, n=6), Kidney ischemia-reperfusion (K-IR, n=6), And kidney ischemia reperfusion+Ambroxol (K-IR+AMB, n=6). In K-IR+AMB group, Ambroxol (30 mg/kg) was administered orally 30 min before the ischemia period. K-IR and K-IR+AMB groups underwent 45 min of kidney ischemia followed by a 6-hour reperfusion period. At the end of the reperfusion period, blood and kidney tissue samples were collected after euthanasia. From the blood samples, BUN and creatinine  levels were determined to assess kidney function, and TNF-α and IL-1β concentrations were evaluated to determine inflammatory response.  While serum BUN, creatinine  activities, and TNF-α and IL-1β concentrations were higher in both IR groups compared with the CTR group, these values were found to be lower in the K-IR+AMB group compared with the K-IR group. Histopathological examination revealed that interstitial edema and desquamation of tubular cells in the K-IR group were more severe than in the K-IR+AMB group.  Ambroxol treatment alleviated the production of pro-inflammatory cytokines and the harmful cellular effects in the tubular cells.