Iranian Journal of Basic Medical Sciences
Volume:25 Issue: 2, Feb 2022

Immunotherapy is a novel preference for the treatment of various complex diseases. Considering the application of varying agents for suppression or activation of the immune system, immunogene therapy was confirmed to stand as a proper alternative for other immunotherapeutic strategies due to its capability in targeting cells with more specificity that leads to controlling the expression of therapeutic genes. This method facilitates the local and single-dose application of most gene therapies that result in the usage of high therapeutic doses with a low risk of systemic side effects while being cost-efficient in long-term administrations. However, the existing barriers between the administration site and cell nucleus limited the clinical uses of genetic materials. These challenges can be overcome through the promising method of exerting non-carriers with high stability, low toxicity/immunogenicity, and simple modifications. In this study, we attempted to review the potential of nanoparticle application throughout the immunogene therapy of different diseases including cancer, microbial diseases, allergies, inflammatory bowel disease, rheumatoid arthritis, and respiratory infections. We included the outline of some challenges and opportunities in regards to the delivery of genetic materials that are based on nano-systems through immunotherapy of these disorders. Next to the promising future of these vectors, more detailed analyses are required to overcome the current limitations in clinical approaches.

Calcium dobesilate (CaD) has anti-oxidant, anti-inflammatory, and anti-apoptotic effects. In this study, the protective effects of CaD against hepatorenal damage induced by carbon tetrachloride (CCl4) in mice were evaluated. 

Thirty male mice were randomly divided into five groups: Control, CaD 100 mg/kg, CCl4, CCl4+CaD 50 mg/kg, and CCl4+CaD 100 mg/kg. CaD (50 and 100 mg/kg) was administered orally once a day for 4 weeks. The liver and kidney indices (serum creatinine, blood urine nitrogen, alanine aminotransferase, and aspartate aminotransferase levels) were determined. Also, liver and kidney tissue oxidant/anti-oxidant markers (glutathione peroxidase, malondialdehyde, total anti-oxidant capacity, and superoxide dismutase) were measured. Cleaved caspase-3, Bax, cytochrome-c, and Bcl-2 protein levels were measured by immunoblotting method in the liver and kidney tissues. The liver and kidney histopathological changes were evaluated by the Hematoxylin and Eosin (H&E) staining method.

CCl4 induced significant oxidative stress and apoptosis in kidney and liver tissues that was concomitant with histopathological abnormalities in these organs in the CCl4 group versus the control (P<0.05). However, CaD (100 mg/kg) could significantly improve the histopathological change in the liver and kidney tissues of CCl4+CaD 100 mg/kg mice versus the CCl4 group (P<0.05). In addition, CaD (100 mg/kg) attenuated the pro and anti-apoptotic markers in the liver and kidney tissues of CCl4+CaD 100 mg/kg mice versus the CCl4 group (P<0.05).

CaD (100 mg/kg) has a protective effect against hepatorenal injury induced by CCl4 at least via its anti-apoptotic and anti-oxidant properties.

Adipose tissue-derived mesenchymal stromal cells (ASCs) are useful in cell-based therapy.  However, it is well known that diabetes mellitus (DM) alters ASCs’ functionality. The majority of in vitro studies related to ASCs are developed under non-physiological oxygen conditions. Therefore, they may not reflect the full effects of DM on ASCs, in vivo. The main aim of the current study is to identify molecular pathways and underlying biological mechanisms affected by diabetes on ASCs in physiological oxygen conditions.

ASCs derived from healthy (ASCs-C) and diabetic (ASCs-D) rats were expanded under standard culture conditions (21% O2) or cultured in physiological oxygen conditions (3% O2) and characterized. Differential gene expressions (DEGs) of ASCs-D with respect to ASCs-C were identified and analyzed with bioinformatic tools. Protein-protein interaction (PPI) networks, from up- and down-regulated DEGs, were also constructed.

The bioinformatic analysis revealed 1354 up-regulated and 859 down-regulated DEGs in ASCs-D, with 21 and 78 terms over and under-represented, respectively. Terms linked with glycosylation and ribosomes were over-represented and terms related to the activity of RNA-polymerase II and transcription regulation were under-represented. PPI network disclosed RPL11-RPS5 and KDR-VEGFA as the main interactions from up- and down-regulated DEGs, respectively.

These results provide valuable information about gene pathways and underlying molecular mechanisms by which diabetes disturbs ASCs biology in physiological oxygen conditions. Furthermore, they reveal, molecular targets to improve the use of ASCs in autologous transplantation.

Sleep deprivation (SD) has a negative impact on cognitive functions including learning and memory. Many studies have shown that rapid-eye-movement (REM) SD also disrupts memory performance. In this study, we aimed to investigate the effect of multi-epitope Gag-Pol-Env-Tat derived from Human immunodeficiency virus 1 (HIV-1) on REM SD-induced spatial memory impairment with respect to the levels of interleukin-4 (IL-4), interleukin-17 (IL-17), interferon-gamma (IFN-γ), immunoglobulin G1 (IgG1), immunoglobulin G2a (IgG2a), and lymphocyte proliferation in NMRI mice. We used multi-epitope Gag-Pol-Env-Tat derived from HIV-1 because Gag-Pol-Env-Tat immunogen sequence is one of the most sensitive immunogen sequences of HIV-1 that can significantly augment cellular and humoral immune systems, leading to the improvement of cognitive functions.  Morris water maze apparatus was used to assess spatial memory, and multi-platform apparatus was used to induce RSD for 24 hr. Multi-epitope derived from HIV-1 was subcutaneously injected at the dose of 20 µgr/ml, once and fourteen days before RSD.  RSD impaired spatial memory and injection of multi-epitope derived from HIV-1 reversed this effect. RSD decreased IL-4, IgG1, and IgG2a levels, while multi-epitope derived from HIV-1 reversed these effects. Multi-epitope derived from HIV-1 also increased lymphocyte proliferation and decreased IL-17 levels in both control and RSD mice.  Multi-epitope derived from HIV-1 may improve memory performance via induction of anti-inflammatory immune response.

We thought that astaxanthin (ASX) might be a protective agent in oxidative stress damage that develops against ischemia and reperfusion injury in the rat ovary.

The experimental groups consisted of healthy, I (Ischemia), I+ASX50, I+ASX100, I/R (Ischemia/Reperfusion), I/R+ ASX50, and I/R+ ASX100. Vascular clamps were applied to the ovaries for 3 hr to induce ischemia. For the reperfusion groups, the clamps were opened and blood flow was restored to the ovaries for 3 hr. At the end of the experiment, biochemical, histopathological, and immunohistochemical analyses were made from the tissue samples taken.

While MDA levels increased significantly in I and I/R groups, SOD levels decreased. It was found that ASX significantly decreased MDA levels and increased SOD activity in treatment groups depending on the dose. Caspase 3, IL-1 β, and IL-6 expressions were severely increased in ischemia and I/R groups, while the severity of I+ASX50 and I/R+ASX100 immunoreactivity was decreased. While severe hemorrhage areas were observed in I and IR groups, minimal hemorrhage areas were observed in the treatment groups, especially in I/R+ASX100 groups. In addition, inflammatory cells and necrotic cells in the I/R group were not observed in I/R+ASX50 and I/R+ASX100 groups.

As a result, it was determined that ASX has a strong protective role against oxidative damage in the treatment of ovarian ischemia-reperfusion injury.

Methylglyoxal (MG) provokes endoplasmic reticulum (ER) stress in β-cells and triggers pancreatic β-cell dysfunction. Crocin has anti-diabetic properties. The present study investigated whether crocin prevented pancreas damages induced by MG.

Diabetes was induced by MG administration (600 mg/kg/day, PO). On the fourteenth day, after proving hyperglycemia, crocin (15, 30, and 60 mg/kg) and metformin (MT) (150 mg/kg) were used for detoxification of MG until the end of the experiment. The animals were divided into 6 groups: 1) control, 2) diabetic by MG, 3) MG + crocin 15 mg/kg, 4) MG + crocin 30 mg/kg, 5) MG + crocin 60 mg/kg, and 6) MG + MT. The data were analyzed by one-way analysis of variance and significant differences were compared by Tukey and Bonferroni tests (P<0.05). Biochemical assays, antioxidant evaluation, and microRNAs expression associated with ER stress were assessed.

MG induced hyperglycemia, insulin resistance, and dyslipidemia (P<0.001). Crocin and MT significantly ameliorated β-cell function through reduction of fasting blood glucose, malondialdehyde levels (P<0.001), and  significant elevation of anti-oxidant enzyme activity accompanied by regulation of glutathione and glyoxalase1-Nrf2 in MG induced diabetic mice. Crocin and MT significantly down-regulated microRNAs 204, 216b, 192, and 29a expression (P<0.001). Crocin (60 mg/kg) (P<0.01) and MT (P<0.001) could improve diameter of pancreatic islets in MG treated mice. 

Crocin prevents the progression of diabetes through modulating ER stress-associated microRNAs and GLO1 activity with the helpful effects of glutathione and Nrf2.

Frizzled-7, the most common receptor of the Wnt signaling pathway, was significantly over-expressed in gastric (GC) and colorectal (CRC) cancers and stimulated tumorigenesis. The extracellular domain of Fzd7 (sFzd7) as a decoy receptor, could competitively bound with ligands and antagonize the interaction between Fzd7 receptors and Wnt ligands. 

We expressed and purified the extracellular region of Fzd7 including cysteine-rich domain (33 aa–185 aa) from Escherichia coli by chromatography. The effect of sFzd7 was evaluated on AGS gastric and SW480 colon cancer cell lines expressing high levels of Fzd7 receptor. Accordingly, cell viability and apoptosis were measured using MTT and flow cytometry assays, respectively. Real-Time PCR determined the relative expression of the β-catenin and cyclin-D1 genes. 

After three days of treatment with sFzd7, the viability of AGS and SW480 cell lines was decreased in a dose-dependent manner. In addition, sFzd7 at concentrations of 10 and 20 ug/ml increased the rate of apoptosis. Especially at the concentration of 20 ug/ml, the apoptosis rate was remarkably high in AGS (P-value= 0.003) and SW480 cells (P-value= 0.0007). Finally, the expressions of β-catenin (P-value= 0.01) and cyclin-D1 (P-value= 0.02) were obviously decreased in SW480 cells. The same results were obtained in AGS cells, although not statistically significant. 

sFzd7 decoy receptor inhibits tumor cell progression by attenuating the Wnt pathway through inhibiting Fzd7 receptors and Wnt ligand interaction. Hence, sFzd7 can be proposed as a candidate therapy for GC and CRC cells with high levels of Fzd7 expression.

Renal tubular damage is critical pathological feathers of diabetic nephropathy (DN). This study aimed to explore the protective activity and related mechanisms of crocin in renal epithelial cell injury induced by high glucose. Renal tubular epithelial HK-2 cells were cultured with D-glucose to establish an in vitro DN model. Cell viability was evaluated by CCK-8 assay. Apoptosis was detected by Annexin V-FITC kit. Oxidative stress was evaluated by colorimetry. RT-qPCR was carried out to determine the mRNA expressions of NF-E2-related factor 2 (Nrf2) and its pathway genes. Western blot was applied to determine the protein expressions of Nrf2 and related proteins. High glucose (5.5, 30, and 50 mM D-glucose) decreased cell viability at 72 hr, which was attenuated by crocin (25 and 50 μM). Crocin also attenuated the high glucose (30 mM D-glucose) induced apoptosis of HK-2 cells,  decreased MDA content, and increased SOD activity in culture media. Crocin increased mRNA levels of Nrf2, HO-1, and NQO1. Moreover, crocin increased protein expressions of Nrf2, Sirtuin 1 (SIRT1), and p-Akt (Ser473). Inhibition of Nrf2 using siRNA, and inhibitors of SIRT1 (nicotinamide, NAM, 20 μM) and PI3K/Akt (LY294002, 50 μM) all attenuated the protective effect of crocin. Nrf2 siRNA and NAM also partially attenuated the inhibitory effect on oxidative stress and increase in the Nrf2 protein by crocin treatment. Crocin protects renal epithelial cells against injury induced by high glucose, and the mechanism is associated with partial activation of the SIRT1-Nrf2 pathway.

As olanzapine has side effects such as weight gain and metabolic disorders, and alpha-mangostin has been shown to control metabolic disorders, the effects of alpha-mangostin on metabolic disorders induced by olanzapine were investigated in this study. Obesity was induced in female Wistar rats by daily administration of olanzapine (5 mg/kg/day, IP, 14 days). Rats were divided into 6 groups:1) vehicle (control); 2) olanzapine (5 mg/kg/day); 3,4,5) olanzapine+ alpha-mangostin (10, 20, 40 mg/kg/day, IP); 6) alpha-mangostin (40 mg/kg/day). Weight changes were measured every 3 days and food intake was assessed every day. Systolic blood pressure, plasma levels of blood sugar, triglycerides, total cholesterol, HDL, LDL, leptin, oxidative stress markers (MDA, GSH), AMPK, and P-AMPK protein levels in liver tissue were assessed on the last day of the study.  Administration of olanzapine significantly increased weight gain, food intake, blood pressure, triglycerides, LDL, blood sugar, leptin, and MDA in rat liver tissue and also decreased GSH, AMPK, and P-AMPK in liver tissue compared with the control group. Different doses of alpha-mangostin significantly reduced weight gain, food intake, systolic blood pressure, triglycerides, LDL, blood sugar, leptin, and MDA. Also, they significantly increased GSH, AMPK, and P-AMPK in liver tissue compared with the olanzapine group. Olanzapine increases leptin levels, food intake, and weight, induces oxidative stress, decreases the levels of AMPK and P-AMPK proteins in liver tissue, and causes metabolic disorders. But, alpha-mangostin reduces the negative effects of olanzapine by activation of AMPK.

Klebsiella pneumoniae is the common cause of pneumonia in hospitalized patients, particularly in intensive care units (ICU). The infection can transfer by medical equipment such as mechanical ventilators. This study aimed to investigate the molecular typing of the extended-spectrum beta-lactamase-producing K. pneumoniae isolates recovered from Beheshti Hospital, Kashan, Iran. 

K. pneumoniae isolates producing ESBLs have been collected from the samples obtained from Shahid Beheshti hospital, Kashan, Iran. Antimicrobial susceptibility was determined using the Kirby Bauer disk diffusion method. The presence of ESBLs was evaluated using CLSI for ESBL screening by the double-disk diffusion method. Molecular typing was conducted by pulsed-field gel electrophoresis (PFGE).  In total, 89 K. pneumoniae isolates were recovered, of which 47.1% were ESBL producers.

Results showed that all of the clinical and environmental isolates were resistant to ceftriaxone, meropenem, cefazolin, cefotaxime, cephalothin, and piperacillin-tazobactam. All isolates were grouped under four clusters (A-D). The major cluster was related to the C cluster with 22 isolates (19 clinical and 3 environmental). Seventy-two percent of isolates were from the ICU ward. There was no correlation between antibiotic resistance patterns and PFGE clusters (P=0.2).

We observed a common molecular signature among both clinical and environmental K. pneumoniae isolates, indicating a similar genotype and likely a common origin for ESBL producer isolates found in different hospital wards. Therefore, hospitals need to implement an effective infection control system to decrease the spreading of ESBL strains within the hospitals and subsequently the transmission of the infection to patients.

This study aimed to investigate the possible consequences of administering exogenous melatonin as prevention or treatment against tunicamycin-induced endoplasmic reticulum (ER) stress in the testicular tissue of rats. 

In this study, 42 adult Sprague Dawley rats, randomly divided into seven equal groups, were administered intraperitoneal tunicamycin to induce ER stress. Both prophylactic (PMel) and therapeutic melatonin (TMel) groups were administered melatonin for seven days. ER stress in the cell was detected through immunohistochemical and molecular analyses using GPR78 expression. 

Increased oxidant levels and apoptosis rates were shown in testicular tissue because of ER stress. The sections in the melatonin-administered and control groups were similar, with melatonin-administered groups showing an increase in the antioxidant ratio. Histometric examinations revealed both TMel and melatonin applications reduced the diameter of the tubules. However, immunohistochemical and molecular analyses showed that PMel administration decreased the concentration of GRP78 more effectively than TMel. 

Applying melatonin prior to cell damage occurrence can be recommended for its effectiveness in protecting from tunicamycin-induced ER stress.

Tuberculosis affects one-third of the world’s population and leads to a high rate of morbidity and mortality. Bacillus Chalmette–Guerin (BCG) as the only approved vaccine for the Mycobacterium tuberculosis (Mtb) does not show enough protection in the vaccinated population. 

The main aim of this study was to prepare a self-assembled nanomicelle composed from a di-block polymer in which, a di-fusion peptide was the hydrophobic block and polyethylene glycol (PEG) was the hydrophilic block. The micelles were characterized in vitro and in vivo as an antigen delivery system/adjuvant both with and without a prime BCG. 

The micellar nanovaccine was able to elicit good dendritic cell maturation. Nanomicelles could efficiently induce systemic cytokines as well as nasal secretory predominant antibody titers (sIgA). The expression pattern of cytokines indicated the superiority of cellular immunity. Nasal administration of two doses of nanomicelles after a prime subcutaneous administration of BCG induced the highest mucosal and systemic immune responses. 

Based on our results PEG-HspX/EsxS self-assembled nanomicelle is highly immunogenic and can be considered a potential vaccine candidate against Mtb to boost BCG efficiency.

Antimicrobial peptide compounds (AMPs) play important roles in the immune system. They also exhibit significant anti-tumor and antibacterial properties. Most AMPs are cationic and are able to bind bacterial cell membranes through electrostatic affinity. Ib-AMP4 is a plant-derived AMP that exerts rapid bactericidal functions. In the present study, the antibacterial efficiency of the produced recombinant Ib-AMP4 in elimination of Methicillin-resistant Staphylococcus aureus (MRSA) bacterial infection, was investigated under in vitro and in vivo situations. 

The synthesized Escherichia coli codon-optimized gene sequences of the Ib-AMP4 were expressed in E. coli BL21 (DE3) pLysS. The recombinant Ib-AMP4 was purified and refolding conditions were optimized. The antibacterial efficiency of the refolded peptide against MRSA was tested under in vivo and in vitro situations for treatment of skin and systematic infection of MRSA in a mouse model.

Antibacterial assays confirmed the antibacterial function of Ib-AMP4 against MRSA. SEM results proved the destructive effects of applying Ib-AMP4 on MRSA biomembrane. Time-kill curve and growth kinetic assay illustrated rapid antibacterial activity of the produced Ib-AMP4. Moreover, Ib-AMP4 showed significant infection treatment ability in a mouse model and all infected mice receiving Ib-AMP4 protein survived and there was no trace of bacteria in their blood samples.

The results confirmed the rapid antibacterial potential of the produced recombinant Ib-AMP4 to be used for efficient treatment of  MRSA infection.

Oral colonization of Acinetobacter baumannii can lead to infections such as pneumonia and sepsis. We aimed to evaluate oral colonization of hospitalized patients in ICUs and to examine risk factors for oral colonization, molecular epidemiology, and incidence of pneumonia and sepsis.  The study began in February 2021. Oral cultures were taken. The microorganisms were identified by a Maldi-tof MS mass spectrometry device. Colistin resistance genes were investigated by polymerase chain reaction. Clonal relationships were determined by pulsed-field gel electrophoresis.  A. baumannii was found in 21 of 96 patients’ oral cultures. Pneumonia and sepsis due to A. baumannii were detected in 14 and 5 patients, respectively. The mean growth time of A. baumannii from oral cultures was 11.8 days, and the meantime for the occurrence of pneumonia after oral growth was 5.2 days. We determined a plasmid mediated mcr-2 colistin resistance gene in a colistin susceptible A. baumannii strain. It is the first report of the plasmid mediated mcr-2 colistin resistance gene in our country. In total, fourteen different A. baumannii genotypes were determined in PFGE. It was determined that the effects of antibiotic use, oral motor dysfunction, mechanical ventilation, intubation, orogastric tube use, and total parenteral nutrition intake on oral colonization were statistically significant. Oral colonization of A. baumannii is a significant concern in ICUs. We believe that it is important to take oral cultures and follow the risk factors and take infection control measures to prevent oral colonization of resistant isolates in ICUs.

Blood-brain barrier (BBB) permeability is central in multiple sclerosis (MS) pathophysiology, and exercise may improve BBB integrity. The current study investigated the prophylactic and/ or therapeutic role of aerobic exercise (EX) training on BBB integrity in experimental autoimmune encephalomyelitis (EAE). 

Forty female Lewis rats were randomly divided into four groups. The experimental groups included: no-EAE induction+ no-exercise (no-EAE+ no-EX), no-EAE induction+ exercise (no-EAE+EX), EAE induction+ no-exercise (EAE+ no-EX), and EAE induction+ exercise (EAE+EX). The no-EAE+EX and EAE+EX groups performed six weeks of progressive aerobic exercise training. GFAP, angiopoietin 1 (Ang-1) expression, tight-junction (TJ) proteins of claudin-5 and occludin were measured as components of BBB integrity and the rate of neuronal apoptosis was evaluated in hippocampi. 

A significant increase in GFAP and Ang-1 expression (P<0.001) and conversely a down-regulation in TJ proteins (P<0.05) was found in the brains of the no-EAE+EX group compared with the no-EAE+ no-EX group. The expression of GFAP and Ang-1 proteins significantly increased in the hippocampi of the EAE+ no-EX group (P<0.001), whereas aerobic training (in the EAE+EX group) meaningfully reversed such increases (P<0.001). Besides, down-regulated TJ proteins and increased neuronal apoptosis induced by EAE induction (EAE+ no-EX group) were restored and reduced, respectively, by aerobic training in the CNS of the EAE+EX group (P<0.001). 

The provision of a six-week treadmill aerobic training buffered the detrimental effects of EAE on BBB integrity and consequently neuronal apoptosis.

High-intensity interval training (HIIT) is a shape of interval training that provides ameliorated athletic capacity and has a good effect on health. Resveratrol is a natural polyphenol abundant in grapes and red wine and has been demonstrated to apply various useful health impacts on the body. This research aimed to evaluate the interactive effects of swimming HIIT and resveratrol consumption on SIRTs 3 & 4, NAD+/NADH, AMPK and SOD2 expression in aged rats.

In total, forty-five old male albino rats (Wistar) with the age of twenty months were allocated into 5 groups randomly. Control group (Ctrl), Swimming HIIT group (Ex: Exercise), Swimming HIIT with Resveratrol consumption group (R+Ex), Resveratrol consumption group (R) and solvent of resveratrol consumption group (vehicle). R+Ex group accomplished the exercise and consumed resveratrol (10 mg/kg/day, gavage) for 6 weeks.

HIIT & resveratrol significantly increased NAD+/NADH, SOD 2 and AMPK in the aged rats. HIIT increased SIRT3, but resveratrol reduced it. As for SIRT4, HIIT decreased it, while resveratrol positively affected it.

Resveratrol and HIIT, especially their combination, have anti-oxidant and anti-aging effects on the hippocampus of old rats.

This study was conducted to examine the therapeutic effects of lavender oil (LO) against oxygen-glucose deprivation (OGD)-induced injury in vitro model.

In this study, the OGD model was induced in the H9C2 cell line, and then the cells were treated with LO (10, 100, 1000, and 10000 μg/ml). The anti-inflammatory activity of LO (Jak2/Stat3) was evaluated by immunocytochemical assay. Likewise, the p-ERK/ERK level was measured by western blotting.

Compared with only the OGD-induced injury model, cell survival increased after treatment with LO. Our results showed that 100 μg/ml of LO significantly decreased the expression of Jak2/Stat3 and the apoptotic activity 72 hr after reperfusion compared with the control group. Likewise, significant increases were observed in p-ERK/ERK in LO-treated groups. 

Collectively, these findings confirm that LO can be a good candidate to reduce OGD-induced injury in the H9C2 cell line through targeting Jak2/Stat3 and ERK pathways.