Publication: Nörotoksinler ile oluşturulan parkinson hastalığı modelinde sodyum-glukoz ko-transporter 2 inhibisyonunun etkilerinin araştırılması
Abstract
Amaç: Çalışmamızdaki amacımız, zebra balıklarında oluşturulan Parkinson hastalığı (PH) modelinde sodyum-glukoz kotransporter 2 inhibitörü olan empagliflozinin olası ketojenik, otofajik ve nöroprotektif etkilerinin incelenmesidir. Gereç ve Yöntem: Zebra balıkları Kontrol, Rotenon (ROT), Lipopolisakkarit (LPS), Empagliflozin (EMP), ROT+LPS, ROT+EMP, LPS+EMP, ROT+LPS+EMP olmak üzere sekiz gruba ayrılmıştır. Grupların her hafta ağırlıkları tartılmış, kaydedilmiş ve maruziyet süresi bitiminde davranış analizleri yapılmıştır. Ardından grupların kan glukoz düzeyleri ölçülmüş, beyin ve hepatopankreas dokuları alınmıştır. Dokularda biyokimyasal parametreler spektrofotometrik yöntem ile, nörotransmitter ölçümü LC-MS/ MS yöntemi ile, keton cisimlerinin düzeyleri kolorimetrik yöntem ile belirlenmiştir. PH ile ilişkili gen ifadeleri RT-PCR yönetimi ile, uygulanan maruziyetlerin gen ekspresyonları üzerindeki etkileri mikroarray yöntemi ile belirlenmiştir. Tirozin hidroksilaz (th) ve lenfosit sitozolik protein 1 (lcp1) ifadelerinin değişimi immunohistokimyasal yöntem ile belirlenmiştir. Bulgular: Çalışmamızda EMP, dört hafta sonunda vücut ağırlıklarında azalmaya sebep olmuştur. Y-maze analizinde spontan değişim performansı ROT ve LPS verilen gruplarda azalırken, EMP uygulaması ile ROT ve ROT+LPS gruplarında artış gözlenmiştir. EMP uygulaması oksidatif stresin azalmasına yardımcı olmuştur. Beyin ve hepatopankareasta yapılan keton cisimleri analizinde EMP verilen grupta artış gözlenmiştir. EMP etkisi ile DOPAC/ DA oranı artmıştır. Mikroarray analizinde, EMP’nin otofajik yolaklarda etkili olduğu görülmüştür. RT-PCR çalışmasında EMP’nin beyin dokusunda nöroinflamasyonu azaltıcı etkiye sahip olduğu, sirt1 genin ekspresyonunu arttırdığı ve pink1 ve lrrk2 genlerinin ekspresyonunu azalttığı görülmüştür. Hepatopankreas dokusunda ise hmgcs1 geninin ekspresyonu azalmıştır. İmmünohistolojik incelemede EMP’nin th ifadesini arttırırken lcp1 ifadesinin azalmasını sağlayarak nöroinflamasyonu azalttığı bulunmuştur. Sonuç: Çalışmamızın sonuçlarına göre zebra balıklarında ROT ve LPS ile oluşturulan PH modelinde EMP’nin olumlu etkileri olduğu gözlemlenmiştir.
Objective: The aim of our study is to we explore the possible ketogenic, autophagic and neuroprotective effects of empagliflozin, a sodium-glucose cotransporter 2 inhibitor, in a zebrafish Parkinson's disease (PD) model. Materials and Methods: Zebrafish were divided into eight groups as Control, Rotenone (ROT), Lipopolysaccharide (LPS), Empagliflozin (EMP), ROT+LPS, ROT+EMP, LPS+EMP, ROT+LPS+EMP. Every week, the weights of the groups were measured and recorded. The behavioral assessments were completed after the exposure period. Then, the blood glucose levels of the groups were measured, and the brain and hepatopancreas tissues were taken. Afterwards, biochemical parameters were determined by spectrophotometric method, neurotransmitter measurement was determined by LC-MS/ MS method, and ketone body levels were determined by colorimetric method. PD-related gene expressions were determined by RT-PCR, and the effects of applied exposures on gene expressions were determined by microarray method. Immunohistochemical analysis was used to detect the alterations in tyrosine hydroxylase (th) and lymphocyte cytosolic protein 1 (lcp1) expression. Results: EMP caused a decrease in body weights at the end of four weeks. In the Y-maze analysis, while the spontaneous change performance decreased in the ROT and LPS groups, the exposure of EMP caused an increase and decreased oxidative stress in these groups. EMP group had higher levels of ketone bodies in the brain and hepatopancreas. The DOPAC/ DA ratio increased with the effect of EMP. Microarray analysis showed that EMP acted on autophagic pathways. In the RT-PCR study, it was shown that EMP reduces neuroinflammation in brain tissue, enhances sirt1 gene expression, and decreases pink1 and lrrk2 gene expression. In hepatopancreatic tissue, the hmgcs1 gene's expression was downregulated. In the immunohistological examination, the expression of th increased in the groups given EMP and decreased neuroinflammation was found through reduced lcp1 expression. Conclusion: According to the results of our study, it has been observed that EMP has positive effects in the PH model generated with ROT and LPS in zebrafish.
Objective: The aim of our study is to we explore the possible ketogenic, autophagic and neuroprotective effects of empagliflozin, a sodium-glucose cotransporter 2 inhibitor, in a zebrafish Parkinson's disease (PD) model. Materials and Methods: Zebrafish were divided into eight groups as Control, Rotenone (ROT), Lipopolysaccharide (LPS), Empagliflozin (EMP), ROT+LPS, ROT+EMP, LPS+EMP, ROT+LPS+EMP. Every week, the weights of the groups were measured and recorded. The behavioral assessments were completed after the exposure period. Then, the blood glucose levels of the groups were measured, and the brain and hepatopancreas tissues were taken. Afterwards, biochemical parameters were determined by spectrophotometric method, neurotransmitter measurement was determined by LC-MS/ MS method, and ketone body levels were determined by colorimetric method. PD-related gene expressions were determined by RT-PCR, and the effects of applied exposures on gene expressions were determined by microarray method. Immunohistochemical analysis was used to detect the alterations in tyrosine hydroxylase (th) and lymphocyte cytosolic protein 1 (lcp1) expression. Results: EMP caused a decrease in body weights at the end of four weeks. In the Y-maze analysis, while the spontaneous change performance decreased in the ROT and LPS groups, the exposure of EMP caused an increase and decreased oxidative stress in these groups. EMP group had higher levels of ketone bodies in the brain and hepatopancreas. The DOPAC/ DA ratio increased with the effect of EMP. Microarray analysis showed that EMP acted on autophagic pathways. In the RT-PCR study, it was shown that EMP reduces neuroinflammation in brain tissue, enhances sirt1 gene expression, and decreases pink1 and lrrk2 gene expression. In hepatopancreatic tissue, the hmgcs1 gene's expression was downregulated. In the immunohistological examination, the expression of th increased in the groups given EMP and decreased neuroinflammation was found through reduced lcp1 expression. Conclusion: According to the results of our study, it has been observed that EMP has positive effects in the PH model generated with ROT and LPS in zebrafish.