Andrographolide induced heme oxygenase-1 expression in MSC-like cells isolated from rat bone marrow exposed to environmental stress.

BACKGROUND AND OBJECTIVES: Mesenchymal stem cells (MSC-like cells) are the most important stem cells that are used in transplantation clinically in various applications. The survival rate of MSC-like cells is strongly reduced due to adverse conditions in the microenvironment of transplantation, including environmental stress. Heme oxygenase-1 (HO-1) is a member of the heat shock protein, as well as a stress-induced enzyme, present throughout the body. The present study was conducted to investigate the effect of andrographolide, an active derivative from andrographolide paniculate, on HO-1 expression in mesenchymal stem cells derived from rat bone marrow. MATERIALS AND METHODS: The rat bone marrow-derived mesenchymal stem cells (BMSC-like cells) were extracted and proliferated in several passages. The identity of MSC-like cells was confirmed by morphological observations and differential tests. The flow cytometry method was used to verify the MSC-specific markers. Isolated MSC-like cells were treated with different concentrations of andrographolide and then exposed to environmental stress. Cell viability was assessed using the MTT colorimetric assay. A real-time PCR technique was employed to evaluate the expression level of HO-1 in the treated MSC-like cells. RESULTS: Isolated MSC-like cells demonstrated fibroblast-like morphology. These cells in different culture mediums differentiated into osteocytes and adipocytes and were identified using alizarin red and oil red staining, respectively. As well, MSC-like cells were verified by the detection of CD105 surface antigen and the absence of CD14 and CD45 antigens. The results of the MTT assay showed that the pre-treatment of MSC-like cells with andrographolide concentration independently increased the viability and resistance of these cells to environmental stress caused by hydrogen peroxide and serum deprivation (SD). Real-time PCR findings indicated a significant increase in HO-1 gene expression in the andrographolide-receiving groups (p < 0.01). CONCLUSION: Our results suggest that andrographolide creates a promising strategy for enhancing the quality of cell therapy by increasing the resistance of MSC-like cells to environmental stress and inducing the expression of HO-1.

Comparison of the effects of zinc oxide and zinc oxide nanoparticles on the expression of hepcidin gene in rat liver.

OBJECTIVES: Nanoparticles have special properties, such as increased intestinal absorption, permeability, and so on. Zinc oxide (ZnO) nanoparticles have medical applications such as using in drug production. Studies of ZnO nanoparticles have shown the role of these particles in reducing or increasing the genes expression. Given the important role of hepcidin in the development of anemia and iron overload diseases, this study investigated the effect of ZnO nanoparticles on the hepatic expression of the hepcidin gene to help find a way to treat these diseases. METHODS: In this experimental study, 24 male Westar rats were divided into three groups: control, ZnO treating group and ZnO nanoparticle treating group. Both ZnO and ZnO nanoparticles were injected with 50 mg/kg body weight for 14 days. At the end, serums were collected and iron, ferritin and IL-6 levels were measured. Expression of the hepcidin gene was done by Real Time PCR. RESULTS: ZnO and the ZnO nanoparticle significantly increased the expression of the hepcidin gene relative to the control group. The increase in expression of the hepcidin gene in ZnO nanoparticles was more significant than in the ZnO. CONCLUSION: ZnO nanoparticles led to significant increase in expression of the hepcidin gene.

Genetic variants within Ninjurin 2 gene are associated with risk of ischemic stroke in Iranian population.

Previous genetic and epidemiological studies have shown the contribution of genetic factors in conferring the risk of ischemic stroke. Among the acknowledged risk factors of stroke are the single nucleotide polymorphisms (SNPs) near Ninjurin 2 (NINJ2) gene which encodes a surface adhesion protein. In the current study, we investigated the role of two SNPs near this gene in ischemic stroke in Iranian population. The frequency of the A allele of the rs11833579 was significantly lower in cases compared with controls (OR (95% CI) = 0.68 (0.54-0.86), adjusted P value = 0.002). The rs11833579 was significantly associated with risk of stroke in co-dominant (AA vs. GG: OR (95% CI) = 0.39 (0.23-0.66), adjusted P value = 0.003) and recessive (OR (95% CI) = 0.44 (0.27-0.72), adjusted P value = 0.001) models. The rs3809263 was associated with risk of stroke in dominant model (OR (95% CI) = 1.5 (1.09-2.06), adjusted P value = 0.02). The A C haplotype (rs11833579 and rs3809263) decreased the risk of stroke (OR (95% CI) = 0.72 (0.57-0.91), adjusted P value = 0.03), while the G T haplotype conferred susceptibility to stroke (OR (95% CI) = 1.42 (1.11-1.82), adjusted P value = 0.02). Consequently, the present case-control study supports the role of NINJ2 as a risk locus for ischemic stroke in Iranian population.

Pharmacological Notch pathway inhibition leads to cell cycle arrest and stimulates ascl1 and neurogenin2 genes expression in dental pulp stem cells-derived neurospheres.

OBJECTIVE: Human dental pulp-derived stem cells (hDPSCs) are becoming an attractive source for cell-based neurorestorative therapies. As such, it is important to understand the molecular mechanisms that regulate the differentiation of hDPSCs toward the neuronal fate. Notch signaling plays key roles in neural stem/progenitor cells (NS/PCs) maintenance and prevention of their differentiation. The aim of this study was to address the effects of Notch signaling inhibition on neurosphere formation of hDPSCs and neuronal differentiation of hDPSCs-neurospheres. RESULTS: hDPSCs were isolated from third molar teeth. The cultivated hDPSCs highly expressed CD90 and CD44 and minimally presented CD34 and CD45 surface markers. The osteo/adipogenic differentiation of hDPSCs was documented. hDPSCs were cultured in neural induction medium and N-[N-(3,5-difluorophenacetyl-L-alanyl)]-Sphenylglycine t-butyl ester (DAPT) was applied to impede Notch signaling during transformation into spheres or on the formed neurospheres. Our results showed that the size and number of neurospheres decreased and the expression profile of nestin, sox1 and pax6 genes reduced provided DAPT. Treatment of the formed neurospheres with DAPT resulted in the cleaved Notch1 reduction, G0/G1 arrest and a decline in L-lactate production. DAPT significantly reduced hes1 and hey1 genes, while ascl1 and neurogenin2 expressions augmented. The number of MAP2 positive cells improved in the DAPT-treated group. CONCLUSIONS: Our findings demonstrated the Notch activity in hDPSCs-neurospheres. DAPT treatment positively regulated proneural genes expression and increased neuronal-like differentiation.