ABSTRACT
Background: Mesenchymal stem cells [MSCs] have been recently received increasing attention for cell-based therapy, especially in regenerative medicine. However, the low survival rate of these cells restricts their therapeutic applications. It is hypothesized that autophagy might play an important role in cellular homeostasis and survival. This study aims to investigate the regenerative potentials of autophagy-modulated MSCs for the treatment of acute liver failure [ALF] in mice
Methods: ALF was induced in mice by intraperitoneal injection of 1.5 ml/kg carbon tetrachloride. Mice were intravenously infused with MSCs, which were suppressed in their autophagy pathway. Blood and liver samples were collected at different intervals [24, 48 and 72 h] after the transplantation of MSCs. Both the liver enzymes and tissue necrosis levels were evaluated using biochemical and histopathological assessments. The survival rate of the transplanted mice was also recorded during one week
Results: Biochemical and pathological results indicated that 1.5 ml/kg carbon tetrachloride induces ALF in mice. A significant reduction of liver enzymes and necrosis score were observed in autophagy-modulated MSC-transplanted mice compared to sham [with no cell therapy] after 24 h. After 72 h, liver enzymes reached their normal levels in mice transplanted with autophagy-suppressed MSCs. Interestingly, normal histology without necrosis was also observed
Conclusion: Autophagy suppression in MSCs ameliorates their liver regeneration potentials due to paracrine effects and might be suggested as a new strategy for the improvement of cell therapy in ALF
ABSTRACT
Objective: In spite of accumulating information about pathological aspects of sulfur mustard [SM], the precise mechanism responsible for its effects is not well understood. Circulating microRNAs [miRNAs] are promising biomarkers for disease diagnosis and prognosis. Accurate normalization using appropriate reference genes, is a critical step in miRNA expression studies. In this study, we aimed to identify appropriate reference gene for microRNA quantification in serum samples of SM victims
Materials and Methods: In this case and control experimental study, using quantitative real-time polymerase chain reaction [qRT-PCR], we evaluated the suitability of a panel of small RNAs including SNORD38B, SNORD49A, U6, 5S rRNA, miR-423-3p, miR-191, miR-16 and miR-103 in sera of 28 SM-exposed veterans of Iran-Iraq war [1980-1988] and 15 matched control volunteers. Different statistical algorithms including geNorm, Normfinder, best-keeper and comparative delta-quantification cycle [Cq] method were employed to find the least variable reference gene
Results: miR-423-3p was identified as the most stably expressed reference gene, and miR- 103 and miR-16 ranked after that
Conclusion: We demonstrate that non-miRNA reference genes have the least stability in serum samples and that some house-keeping miRNAs may be used as more reliable reference genes for miRNAs in serum. In addition, using the geometric mean of two reference genes could increase the reliability of the normalizers
ABSTRACT
Trauma is one of the causes of peripheral nerve injuries. Free radicals increase after tissue damages. Usually free radicals are scavenged and detoxified by antioxidants. In this study we assessed antioxidative role of NGAL [Neutrophil gelatinase associated lipocalin=Lipocalin2] molecule on crushed sciatic nerves of rats. In this study 40 rat's sciatic nerves were crushed and then at day 1 and 3 and week 1, 3, 5 after injury the sciatic nerve of rats which suffering crushed were extracted. First of all total mRNA of samples was extracted and then cDNA was synthesized. The expression of NGAL gene was confirmed by using semiquantitative RT-PCR analysis. For immunohistochemistry analysis, the samples were fixed in praformaldehyde and cut in 20 micrometer slices by cryostat. The results showed that the expression of lipocalin 2 was significantly higher in day 1, day 3 and week 1 in crushed sciatic nerves in compare with the intact [non-injured] nerves [p<0.0001]. Immunohistochemistry results also confirmed the protein expression of these genes. The sciatic nerve injuries cause oxidative stress which followed by crush injury. In this research NGAL molecule showed maximum up-regulation in degeneration process after injury when oxidative stress have maximum peak. In this project for the first time we demonstrated the role of this molecule in degeneration and regeneration process of peripheral nerves and with more studies it may suggest as new complemental treatment
ABSTRACT
Airway remodelling is characterized by the thickening and reorganization of the airways seen in mustard lung patients. Mustard lung is the general description for the chronic obstructive pulmonary disease induced by sulfur mustard[SM]. Pulmonary disease was diagnosed as the most important disorder in individuals that had been exposed to sulfur mustard. Sulfur mustard is a chemical warfare agent developed during Wars. Iraqi forces frequently used it against Iranian during Iran -Iraq in the 1980-1988. Peribronchial fibrosis result from airway remodeling that include excess of collagen of extracellular matrix deposition in the airway wall. Some of Smads families in association with TGF-are involved in airway remodeling due to lung fibrosis. In the present study we compared the mRNA expression of Smad2, Smad3, and Smad4 and Smad7 genes in airway wall biopsies of chemical-injured patients with non-injured patients as control. We used airway wall biopsies of ten unexposed patients and fifteen SM-induced patients. Smads expression was evaluated by RT-PCR followed by bands densitometry. Expression levels of Smad3 and Smad4 in SM exposed patients were upregulated but Smad2 and Smad7 was not significantly altered. Our results revealed that Smad3, and 4 may be involved in airway remodeling process in SM induced patients by activation of TGF-. Smad pathway is the most represented signaling mechanism for airway remodeling and peribronchial fibrosis. The complex of Smads in the nucleus affects a series of genes that results in peribronchial fibrosis in SM-induced patients
ABSTRACT
Neutrophil gelatinase-associated lipocalin [NGAL/Lcn2], comprise a group of small extracellular proteins with a common beta-sheet-dominated 3-dimensional structure. In the past, it was assumed that the predominant role of lipocalin was acting as transport proteins. Recently it has been found that oxidative stress induces Lcn2 expression. It has been also proved that electromagnetic field [EMF] produces reactive oxygen species [ROS] in different tissues. Expression of Lcn2 following exposure to electromagnetic field has been investigated in this study. Balb/c mice [8 weeks old] were exposed to 3 mT, 50 HZ EMF for 2 months, 4 hr/day. Afterwards, the mice were sacrificed by cervical dislocation and livers were removed. The liver specimens were stained with Haematoxylin-Eosin [H and E] and analyzed under an optical microscope. Total RNA was extracted from liver and reverse transcription was performed by SuperScript III reverse transcriptase with 1 micro g of total RNA. Assessment of Lcn2 expression was performed by semiquantitative and real time-PCR. The light microscopic studies revealed that the number of lymphocyte cells was increased compared to control and dilation of sinosoids was observed in the liver. Lcn2 was up-regulated in the mice exposed to EMF both in mRNA and protein levels. To the extent of our knowledge, this is the first report dealing with up-regulation of Lcn2 in liver after exposure to EMF. The up-regulation might be a compensatory response that involves cell defense pathways and protective effects against ROS. However, further and complementary studies are required in this regards