Title Izražaj sigma 1 receptora u bubrezima dijabetičnih štakora
Title (english) Expression of the sigma 1 receptor in the kidneys of diabetic rats
Author Ivana Milardović
Mentor Natalija Filipović (mentor)
Committee member Katarina Vukojević (predsjednik povjerenstva)
Committee member Mirna Saraga-Babić (član povjerenstva)
Committee member Natalija Filipović (član povjerenstva)
Granter University of Split School of Medicine (Hystology and embryology) Split
Defense date and country 2019-02-15, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Pharmacy Pharmacy
Abstract Sigma receptori su izvorno otkriveni u živčanom sustavu, ali ih nalazimo i u različitim drugim tkivima, uključujući bubreg. Dosadašnja istraživanja ukazuju na renoprotektivno djelovanje σ1R koje je većinom posredovano modulacijom signalnog puta dušikovog oksida. S obzirom na važnost ovog signalnog puta u patogenezi dijabetičke nefropatije za pretpostaviti je da i σ1R imaju važnu ulogu u patogenezi ovoga stanja. Svrha istraživanja je bila utvrditi promjene izražaja σ1R u bubrezima, koje bi posredno ukazivale na njihovu eventualnu ulogu u patogenezi dijabetičke nefropatije. Istraživanje je provedeno na Sprague Dawley štakorima. Model šećerne bolesti tipa 1 izazvan je injekcijom streptozotocin (55 mg/kg). U pokus su uključeni štakori kojima je 4. dan nakon indukcije šećerne bolesti izmjerena koncentracija glukoze u krvi >16.5 mmol/L. Nakon 2 tjedna odnosno 2 mjeseca životinje su eutanazirane te je uzet uzorak tkiva bubrega. Tkivo je pripremljeno te su napravljeni parafinski rezovi, te obojeni imunofluorescencijski i histološkom tehnikom PAS. Obojeni rezovi fotografirani su te su dobivene mikrofotografije analizirane korištenjem ImageJ. Za statističku analizu korištena je jednosmjerna analiza varijance (ANOVA) i Tukey-ev test za višestruke usporedbe (GraphPad Software, La Jolla, CA, SAD). Kod dijabetične skupine štakora 2 mjeseca nakon injekcije STZ-a vidljive su patološke promjene u bubregu. Te promjene su: ekspanzija mezangija, tubularna skleroza, nakupljanje hijalinog materijala u stijenkama krvnih žila i vakuolizacije stanica distalnih tubula. Analiziran je broj σ1R imunoreaktivnih distalnih tubula i uočeno da u kontrolnim skupinama broj pozitivnih distalnih tubula opada s vremenom, dok u dijabetičnim skupinama raste (oba P<0,001). Broj pozitivnih tubula bio je značajno niži u dijabetičnoj skupini 2 tjedna nakon injekcije STZ-a u usporedbi s odgovarajućom kontrolnom skupinom (P<0,01); dok je broj σ1R pozitivnih tubula bio statistički značajno veći u dijabetičnoj skupini 2 mjeseca nakon indukcije šećerne bolesti u usporedbi s odgovarajućom kontrolnom skupinom (P<0,001). Površina σ1R imunoreaktivnosti bila je statistički značajno veća u dijabetičnoj skupini 2 mjeseca nakon indukcije šećere bolesti u usporedbi s kontrolnom skupinom, ali i u usporedbi s dijabetičnom skupinom 2 tjedna nakon indukcije šećerne bolesti (oba P<0,01). U vakuoliziranim stanicama distalnih tubula dijabetičnih štakora 2 mjeseca nakon injekcije STZ-a uočena je kolokalizaciju σ1R i upalnog čimbenika rasta TGFβ. Našim istraživanjem utvrdili smo najjači izražaj σ1R u stanicama distalnih tubula bubrega štakora. Dinamika izražaja ovih receptora posredno ukazuje na njihovu ulogu u postnatalnom sazrijevanju bubrega štakora te u razvoju dijabetičke nefropatije, posebice oštećenja distalnih tubula.
Abstract (english) Sigma receptors are originally detected in the nervous system, but they are also found in various other tissues, including the kidney. Previous studies indicate the renoprotective activity of σ1R, which is mostly mediated by modulation of the nitric oxide signaling pathway. Considering the importance of this signaling pathway in the pathogenesis of diabetic nephropathy, it could be assumed that σ1R have an important role in the pathogenesis of this condition. The purpose of the study was to investigate changes in the expression of σ1R in the kidneys, which could indicate their possible role in the pathogenesis of diabetic nephropathy. The research was carried out on Sprague Dawley rats. The type 1 diabetes mellitus was induced by the injection of streptozotocin (55 mg/kg). Rats with blood glucose level >16.5 mmol/L on the 4th day following the induction of diabetes were included in the experiment. The animals were sacrificed 2 weeks or 2 months after diabetes induction and a kidney tissue sample was harvested. The tissue was embedded in paraffin and 5 µm thick section were made, and stained with histological technique PAS and immunofluorescence method. Stained sections were photographed, and the resulting images were analyzed using ImageJ. For statistical analysis, a one-way variance analysis (ANOVA) and a Tukey test for multiple comparisons were used (GraphPad Software, La Jolla, CA, USA). In the kidney of the diabetic group of rats 2 months after injection of STZ, pathological changes characteristic for diabetes nephropathy were apperent, including: the expansion of mesangium, tubular sclerosis, the accumulation of hyaline material in the walls of the blood vessels and the vacuolization of distal tubule cells. The number of σ1R immunoreactive distal tubules in the control gruops decreased with time, while in diabetic groups it increased (both P<0.001). The number of immunoreactive tubules was significantly lower in the diabetic group 2 weeks after STZ injection when compared to the corresponding control group (P<0.01); while the number of σ1R positive tubules was significantly higher in the diabetic group 2 months after the induction of diabetes in comparison to the corresponding control group (P<0.001). The area of σ1R immunoreactivity was significantly higher in the diabetic group 2 months after the induction of diabetes compared to the control group, but also in comparison with the diabetic group 2 weeks after the induction of diabetes (both P<0.01). In vacuolated distal tubule cells of diabetic rats 2 months after injection of STZ, colocalization of σ1R and inflammatory growth factor TGFβ was observed. Our research has shown the strongest expression σ1R in the distal tubules in kidney of the rats. The dynamics of the expression of these receptors indicate their role in postnatal etiology of kidney maturation and in the development of diabetic nephropathy, in particular in the damage of distal tubules.
Keywords
Model šećerne bolesti
Štakori Sprague-Dawley
Dijabetičke nefropatije
Sigma receptori
Keywords (english)
Diabetes Mellitus Experimental
Rats Sprague-Dawley
Diabetic Nephropathies
Receptors sigma
Language croatian
URN:NBN urn:nbn:hr:171:884113
Study programme Title: Pharmacy Study programme type: university Study level: integrated undergraduate and graduate Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
Type of resource Text
File origin Born digital
Access conditions Open access
Terms of use
Created on 2019-03-08 10:39:20