ABSTRACT
BACKGROUND: Kidney failure is a debilitating disorder with limited treatment options. The kidney-protective effects of stem cells have been vastly investigated and promising results have been achieved with various sources of stem cells. However, in spite of beneficial effects on other disease models, the renoprotective potential of human cord blood-derived unrestricted somatic stem cells (USSC) has not been examined so far. METHODS: In the present study, acute kidney failure was induced in female nude mice and the effect of USSC transplantation on kidney function and structure was assessed. Furthermore, the expression of some cytokine genes was examined by real-time PCR. Homing of the transplanted cells into kidneys was assessed by flow cytometry, immunohistochemistry, and real-time PCR. RESULTS: USSC-conditioned medium did not attenuate the in vitro nephrotoxic effects of cisplatin. Transplantation of USSC to nude mice did not protect kidney function and was associated with worsened kidney structural damage. USSC transplantation was also associated with a decline in the renal expression of VEGF-A gene. In spite of these effects, the transplanted cells could not be detected in the kidneys by any of the exploited methods and they were mainly entrapped in the lungs. CONCLUSION: These data indicate that USSC are not suitable for cell therapy in the setting of acute kidney injury. Also, this study shows that these stem cells are able to affect damaged kidneys even if they are not homed there.
Subject(s)
Acute Kidney Injury/surgery , Cord Blood Stem Cell Transplantation , Acute Kidney Injury/chemically induced , Acute Kidney Injury/genetics , Adipocytes/cytology , Adipocytes/metabolism , Animals , Antigens, CD/metabolism , Cell Differentiation , Cell Survival/drug effects , Cisplatin , Cord Blood Stem Cell Transplantation/methods , Culture Media, Conditioned/pharmacology , Cytokines/genetics , Female , Flow Cytometry , Gene Expression , Humans , Kidney/cytology , Kidney/drug effects , Mice , Mice, Inbred BALB C , Mice, Nude , Osteocytes/cytology , Osteocytes/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transplantation, Heterologous , Vascular Endothelial Growth Factor A/geneticsABSTRACT
BACKGROUND: The cytotoxic T lymphocyte associated antigen-4 (CTLA-4) gene, is one of the candidate genes for susceptibility to Graves' disease. This study aimed to investigate the association of Graves' disease and Graves' ophthalmopathy with polymorphisms at position +49 in exon 1 and positions -318 and -1147 in the promoter region of CTLA-4 gene in Iranian patients. METHODS: A total of 205 unrelated Iranian patients with Graves' disease who were referred to the outpatient endocrine clinic of a large university general hospital and 103 sex-matched healthy controls were included in this study. Venous blood was obtained, genomic DNA was extracted by a salting out method, and the polymorphisms at positions +49, -318 and -1147 of the CTLA-4 gene were determined using the PCR-restriction fragment length polymorphism method (PCR-RFLP). Genotype and allele frequencies were determined. RESULTS: The frequency of the G allele at position +49 was significantly higher in patients with Graves' disease than in the control group (27.1% vs. 15.1%, OR=2.096, 95%CI=1.350-3.253 and p<0.01). Significant trends were not seen for the other two polymorphisms studied. In patients with ophthalmopathy, the frequency of the G allele at position +49 was higher than in those without ophthalmopathy (33.8% vs. 20.0%, OR=2.043, 95%CI=1.304-3.202 and p<0.01). CONCLUSION: The results of this study suggest that the G allele at position +49 in exon1 of the CTLA-4 gene is associated with Graves' disease and Graves' ophthalmopathy in Iranian patients.
Subject(s)
Antigens, CD/genetics , Graves Ophthalmopathy/genetics , Polymorphism, Single Nucleotide , Adult , CTLA-4 Antigen , Case-Control Studies , Exons/genetics , Female , Gene Frequency , Genetic Predisposition to Disease , Genotype , Graves Ophthalmopathy/immunology , Humans , Iran , Male , Polymorphism, Restriction Fragment Length , Promoter Regions, Genetic/genetics , T-Lymphocytes, Cytotoxic/physiologyABSTRACT
Cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) is a well-known molecule that regulates T cell activity, with polymorphisms at different regions of this gene having been associated with autoimmune conditions. Pretibial myxedema (PTM), also called Graves' dermopathy, is an autoimmune extrathyroidal manifestation of Graves' disease. We opted to investigate the relationship between three single nucleotide polymorphisms of the CTLA-4 gene (+49A/G, and -318C/T and -1147C/T) and PTM in Iranian patients with Graves' ophthalmopathy (GO). A total of 105 unrelated Iranian patients with GO from the outpatient endocrine clinic of a large university general hospital as well as 103 healthy controls were studied. The genomic DNA was extracted from venous blood samples by a salting out method, and the polymorphisms at +49, -318 and -1147 positions of the CTLA-4 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism method. The GG genotype (OR = 6.000, 95% CI = 1.805-19.940, P = 0.005) and the G allele (OR = 2.653, 95% CI = 1.314-5.357, P = 0.009) at position +49 were significantly associated with PTM in the patient group. The same genotype and allele were also significantly more common among patients (with or without PTM) than controls. No significant association was found for the other two polymorphisms. In conclusion, the +49G allele is associated with increased risk of PTM in patients with GO. Studies with larger sample sizes are needed to confirm the results of the present study.
Subject(s)
Genetic Predisposition to Disease , Graves Disease/genetics , Graves Ophthalmopathy/genetics , Leg Dermatoses/genetics , Myxedema/genetics , Adult , DNA Mutational Analysis , Female , Gene Frequency , Genotype , Graves Disease/complications , Graves Disease/epidemiology , Graves Disease/physiopathology , Graves Ophthalmopathy/complications , Graves Ophthalmopathy/epidemiology , Graves Ophthalmopathy/physiopathology , Humans , Leg Dermatoses/complications , Leg Dermatoses/epidemiology , Leg Dermatoses/physiopathology , Male , Myxedema/complications , Myxedema/epidemiology , Myxedema/physiopathology , Polymorphism, Single Nucleotide , Tibia/anatomy & histologyABSTRACT
Parkinson's disease (PD) is neurodegenerative diseases caused by the loss of dopaminergic neurons in the substantia nigra pars compacta. Stem cell therapy is one of the promising strategies in helping to cure PD. In the present study, human mesenchymal stem cells (MSCs) from eye conjunctiva stromal cells were differentiated into dopaminergic neurons. In this work, after conjunctiva biopsy, mesenchymal stem cells were obtained via adherence to the plastic culture dishes. Then, MSCs were treated with general neurogenic medium containing DMEM supplemented with RA, IBMX and dbcAMP for 6 days. RT-PCR, immunocytochemistry and flow cytometry were used for expression of dopaminergic genes such as TH. As a result, RT-PCR analysis revealed the expression of dopaminergic neuron genes such as TH, Ptx3, Nurr1. Furthermore, immunocytochemistry revealed that the differentiated CJMSCs not only express TH gene, but also express TH protein. Flow cytometry showed that TH, MAP-2 proteins increased significantly as increasing passage number. In conclusion, the reported results indicate that CJMSCs might be a suitable and available source for cell transplantation therapy for the central system diseases such as PD.
