Publication: Zebra balığında oluşturulan epilepsi modelinde valproik asitin ağrı mekanizması üzerine etkileri
Abstract
Amaç: Zebra balıklarında oluşturulan akut epilepsi modelinde, ağrının moleküler mekanizması ve bu mekanizmada endojen opioidlerin epilepsi ile ilişkisinin ortaya çıkarılması, epilepsi tedavisinde antikonvülsan olarak kullanılan valproik asitin ağrı mekanizması üzerine etkilerinin incelenmesidir. Gereç ve Yöntem: Zebra balıkları Kontrol (K), Pentilentetrazol (PTZ), Asetik Asit (AA), Valproik Asit (VPA), PTZ+AA, PTZ+VPA, AA+VPA ve PTZ+AA+VPA grupları ayrılarak davranış analizlerinin ardından beyin dokularında epilepsi, ağrı ve opioidler ile ilişkili genlerin ifadeleri RT-PCR ile incelenmiştir. Oksidan-antioksidan sistem parametreleri spektrofotometrik yöntemle analiz edilmiştir. LC-MS/ MS ile nörotransmitter seviyeleri ölçülmüş ve western blot yöntemiyle protein seviyeleri belirlenmiştir. Bulgular: PTZ grubunda keşif oranı, katedilen mesafe ve ortalama hız azalmış NO, LPO ve GST düzeyleri artmış bunlara karşı SOD düzeyleri ise azalmıştır. Ayrıca inflamasyon belirteçlerinden cox-1, tnf-a, ıl-1β, ıl-6 ifadeleri artmış, beyin dokusunda uyarıcı ve inhibe edici nörotransmitter dengesi bozulmuş, dopamin, nöropinefrin ve seretonin seviyeleri kontrol grubuna kıyasla tüm maruziyet gruplarında anlamlı düzeyde azalmıştır. VPA ön tedavisi ile PTZ’nin neden olduğu nöbetler azalmış, asetelkolinesteraz aktivitesi artmıştır. VPA akut epilepsi, akut ağrı ve epilepside ağrı gruplarında artan inflamasyonu azaltıcı yönde etki etmiştir. Asetik asit grubunda ve epilepside ağrı gruplarında artan vücut eğriliğini azaltıcı yönde etki etmiş, hasar gruplarında azalan asetilkolinesteraz aktivitesini artırmıştır. Ayrıca VPA asetik asit grubunda artan pdyn, pomca, cox-1 ve trpa1 ifadelerini azaltmıştır. Sonuç: Bulgularımız PTZ ile oluşturulan akut epilepsi modelinde asetik asit uygulamasının zebra balıklarında periferik ağrının algılanmasında rol oynayan asit algılayan iyon kanallarının, enkefalinler, oksidatif hasar ve nöroinflamasyonla ilgili parametreler üzerinden ağrıya yanıt ile ilgili mekanizmaların bozulduğunu valproik asit tedavisinin antiinflamatuar ve antinosiseptif etki ettiğini işaret etmektedir.
Objective: To reveal the molecular mechanism of pain and the relationship between endogenous opioids and epilepsy in the acute epilepsy model generated in zebrafish, and to examine the effects of valproic acid, which is used as an anticonvulsant in the treatment of epilepsy, on the pain mechanism. Materials and Methods: Zebrafish Control (K), Pentylenetetrazole (PTZ), Acetic Acid (AA), Valproic Acid (VPA), PTZ+AA, PTZ+VPA, AA+VPA and PTZ+AA+VPA groups after behavioral analysis. The expressions of genes associated with epilepsy, pain and opioids in their tissues were examined by RT-PCR. The oxidant-antioxidant system parameters were analyzed by spectrophotometric method. Neurotransmitter levels were measured by LC-MS/ MS and protein levels were determined by the western blot method. Results: In the PTZ group, exploration rate, distance traveled and average speed decreased, while NO, LPO, and GST levels increased, while SOD levels decreased. In addition, it has been shown that with the increase of oxidative stress, the levels of cox-1, tnf-α, ıl-1β, and ıl-6, which are inflammatory markers, increase, the balance of excitatory and inhibitory neurotransmitters in the brain tissue is disturbed, and dopamine, norepinephrine and serotonin levels are significantly reduced in all exposure groups compared to the control group. Seizures caused by PTZ decreased and acetylcholinesterase activity increased with valproic acid pretreatment. VPA reduced the increased inflammation in acute epilepsy, acute pain and epilepsy pain groups. It decreased the body curvature increased in the acetic acid group and epilepsy pain groups, and increased the acetylcholinesterase activity, which decreased in the injury groups. It was also shown that VPA decreased the increased pdyn, pomca, cox-1 and trpa1 expressions in the acetic acid group. Conclusion: Our findings show that acetic acid application in the acute epilepsy model generated with PTZ disrupts the mechanisms related to pain response through parameters related to acid-sensing ion channels, enkephalins, oxidative damage and neuroinflammation, which play a role in the perception of peripheral pain in zebrafish, and that valproic acid treatment has anti-inflammatory and antinociceptive effects in response to these.
Objective: To reveal the molecular mechanism of pain and the relationship between endogenous opioids and epilepsy in the acute epilepsy model generated in zebrafish, and to examine the effects of valproic acid, which is used as an anticonvulsant in the treatment of epilepsy, on the pain mechanism. Materials and Methods: Zebrafish Control (K), Pentylenetetrazole (PTZ), Acetic Acid (AA), Valproic Acid (VPA), PTZ+AA, PTZ+VPA, AA+VPA and PTZ+AA+VPA groups after behavioral analysis. The expressions of genes associated with epilepsy, pain and opioids in their tissues were examined by RT-PCR. The oxidant-antioxidant system parameters were analyzed by spectrophotometric method. Neurotransmitter levels were measured by LC-MS/ MS and protein levels were determined by the western blot method. Results: In the PTZ group, exploration rate, distance traveled and average speed decreased, while NO, LPO, and GST levels increased, while SOD levels decreased. In addition, it has been shown that with the increase of oxidative stress, the levels of cox-1, tnf-α, ıl-1β, and ıl-6, which are inflammatory markers, increase, the balance of excitatory and inhibitory neurotransmitters in the brain tissue is disturbed, and dopamine, norepinephrine and serotonin levels are significantly reduced in all exposure groups compared to the control group. Seizures caused by PTZ decreased and acetylcholinesterase activity increased with valproic acid pretreatment. VPA reduced the increased inflammation in acute epilepsy, acute pain and epilepsy pain groups. It decreased the body curvature increased in the acetic acid group and epilepsy pain groups, and increased the acetylcholinesterase activity, which decreased in the injury groups. It was also shown that VPA decreased the increased pdyn, pomca, cox-1 and trpa1 expressions in the acetic acid group. Conclusion: Our findings show that acetic acid application in the acute epilepsy model generated with PTZ disrupts the mechanisms related to pain response through parameters related to acid-sensing ion channels, enkephalins, oxidative damage and neuroinflammation, which play a role in the perception of peripheral pain in zebrafish, and that valproic acid treatment has anti-inflammatory and antinociceptive effects in response to these.