ABSTRACT
BACKGROUND AND STUDY AIMS: Spinal cord injury (SCI) is one of the most complicated pathologies that affect active young males. miR-21 primarily regulates several cellular processes. We aimed to elucidate the regulatory role of miR-21 and test methylprednisolone as a disease-modifying agent on experimental SCI tissues. METHODS: A total of 36 8- to 10-week-old adult female Sprague-Dawley rats weighing 250 to 300 g were used. Animals were randomly divided into six groups. Except for groups 1 and 4, the spinal trauma model was applied to all animal groups using the clipping method. In groups 3 and 6, methylprednisolone was given. For real-time polymerase chain reaction (PCR) investigations, rats in groups 1, 2, and 3 were reoperated on after the first postoperative day, whereas those in groups 4, 5, and 6 were reoperated on after postoperative day 7 and spinal cord samples from the laminectomy area were removed for gene expression analysis. Relative gene expression of miR-21, Gfap, Vim, Stat3, Faslg, Pten, Bax, Bcl2, Cox2, and Il6 were determined with quantitative reverse transcription (qRT) PCR. RESULTS: In group 3, the miR-21 expression significantly increased compared with groups 1 and 2. When compared with group 3, a decrease in miR-21 expression was observed in group 6 (p < 0.05). When compared with group 4, group 6 had lower levels of Gfap, Pten, Stat3, and Bax (p < 0.05). CONCLUSIONS: miR-21 supports the beneficial aspects of the body's healing mechanisms following SCI. In the acute phase, the use of methylprednisolone increases miR-21 expression in the early period of trauma. Methylprednisolone increases some astrogliosis and inflammation biomarkers' levels; however, it did not affect the apoptotic biomarkers.
Subject(s)
MicroRNAs , Spinal Cord Injuries , Male , Rats , Female , Animals , Methylprednisolone/pharmacology , Methylprednisolone/therapeutic use , Rats, Sprague-Dawley , bcl-2-Associated X Protein/pharmacology , Spinal Cord Injuries/drug therapy , Spinal Cord , MicroRNAs/genetics , MicroRNAs/pharmacology , Disease Models, AnimalABSTRACT
OBJECTIVE: Different sub-types or genetic variations of Blastocystis sp. are thought to play a role in the differential symptoms caused by the parasite or asymptomatic cases. In this study, it was aimed to clone a fragment of SSUrDNA gene of Blastocystis from a patient in order to define its phylogenetic subtype. METHODS: In this study, DNA isolation from the stool of a Blastocystis infected patient was performed. Blastocystis specific primers were used to amplify a SSUrDNA genomic fragment. The amplified DNA fragment was cloned into a plasmid and sequenced using plasmid specific primers. The obtained DNA sequence was analyzed using BLAST and a phylogenetic tree was constructed using the software MEGA. RESULTS: It was found that the Blastocystis isolate in our study is subtype 3. CONCLUSION: Cloning and sequencing of the target genomic region is suggested for phylogenetic analysis studies.
