Comparison of the protective effects of vanillic and rosmarinic acid on cardiac tissue: Lower limb ischemia-reperfusion model in rats

BACKGROUND: Ischemia/reperfusion injury is one of the most challenging postoperative situations in vascular surgery, both in elective procedures with prolonged clamping time and in delayed emergency cases with vascular occlusion. The inflammatory response that develops during ischemia and the oxygen-free radicals that proliferate during reperfusion have detrimental effects on the brain, heart, and kidneys. In this study, we aimed to compare the effects of vanillic and rosmarinic acid in preventing ischemia/reperfusion injury in a lower limb ischemia-reperfusion model in rats.
METHODS: Thirty-two female Sprague-Dawley rats weighing 185-240 g were randomly divided into four groups of eight animals each. Group 1 was designated as the control, Group 2 as ischemia/reperfusion (I/R), Group 3 as ischemia/reperfusion + vanillic acid (I/R + VA), and Group 4 as ischemia/reperfusion + rosmarinic acid (I/R + RA). In all groups except the control, the infrarenal abdominal aorta was clamped, and 60 minutes of ischemia followed by 120 minutes of reperfusion was performed. Vanillic acid was administered intra-abdominally 15 minutes before the start of reperfusion in Group 3, and rosmarinic acid in Group 4. At the end of the reperfusion phase, blood samples and hearts were collected, and the rats were euthanized. Histopathologically, myofibrillar edema, myocytolysis, focal hemorrhages, and infiltration of polymorphonuclear leukocytes (PMNL) in cardiac tissue were examined. Total antioxidant capacity (TAC), total oxidative status (TOS), oxidative stress index (OSI), 8-OH-deoxyguanosine, lactonase, and arylesterase activity were measured in blood samples.
RESULTS: Myofibrillar edema was most pronounced in the I/R group and less pronounced in the I/R + VA and I/R + RA groups (p=0.005 and p=0.066, respectively). There was no difference between the ischemia/reperfusion groups regarding myocytolysis, focal hemorrhage, and PMNL infiltration (p>0.99). Among all groups, TOS and OSI were lowest in the control group, while TAC was highest. TAC was similar in the I/R + VA and I/R + RA groups but was significantly higher in these two groups than in the I/R group. The lactonase activity in the I/R + VA group was similar to that in the control group but was significantly higher compared to the I/R and I/R + RA groups.
CONCLUSION: Our study shows that vanillic and rosmarinic acids reduce myofibrillar edema in the heart after lower limb ischemia and increase TAC. However, vanillic acid increases the activity of lactonase, an enzyme known for its antioxidant effect, more than rosmarinic acid.

Vanilik ve rosmarinik asidin kalp dokusu üzerindeki koruyucu etkilerinin karþýlaþtýrýlmasý: Sýçanlarda alt ekstremite iskemi-reperfüzyon modeli

AMAÇ: Ýskemi/reperfüzyon hasarý, hem uzun klemp süresi gerektiren elektif prosedürlerde hem de damar týkanýklýðý olan gecikmiþ acil vakalarda damar cerrahisinde görülen en zorlu postoperatif durumlardan biridir. Ýskemi sýrasýnda geliþen enflamatuvar yanýt ve reperfüzyon sýrasýnda artan serbest oksijen radikallerinin beyin, kalp ve böbrekler üzerinde zararlý etkileri vardýr. Bu çalýþmada, sýçanlarda alt ekstremite iskemi/reperfüzyon modelinde vanilik ve rosmarinik asidin iskemi/reperfüzyon hasarýný önlemedeki etkisini karþýlaþtýrmayý amaçladýk.
GEREÇ VE YÖNTEM: Aðýrlýklarý 185-240 g olan, 32 adet diþi Sprague-Dawley ýrký sýçan, rastgele her biri 8 hayvandan oluþan 4 gruba ayrýldý. Grup 1 kontrol, grup 2 iskemi/reperfüzyon (I/R), grup 3 iskemi/reperfüzyon + vanilik asit (I/R + VA) ve grup 4 iskemi/reperfüzyon + rosmarinik asit (I/R +) olarak belirlendi. RA). Kontrol grubu dýþýndaki tüm gruplarda infrarenal abdominal aorta klemplenerek 60 dakika iskemi ve ardýndan 120 dakika reperfüzyon uygulandý. Reperfüzyon baþlamadan 15 dakika önce Grup 3'e vanilik asit, grup 4'e ise rosmarinik asit intra-abdominal olarak uygulandý. Reperfüzyon fazýnýn sonunda kan örnekleri ve kalp dokularý alýnarak sýçanlar sakrifiye edildi. Histopatolojik olarak miyofibriler ödem, miyositoliz, fokal kanama ve kalp dokusunda polimorfonükleer lökosit (PMNL) infiltrasyonu incelendi. Kan örneklerinde total antioksidan kapasite (TAK), total oksidatif durum (TOD), oksidatif stres indeksi (OSÝ), 8-OH-deoksiguanozin, laktonaz ve arilesteraz aktivitesi ölçüldü.
BULGULAR: Miyofibriler ödem en çok Ý/R grubunda belirgindi ve Ý/R + VA ve Ý/R + RA gruplarýnda daha az belirgindi (sýrasýyla, p=0.005 ve p=0.066). Ýskemi/reperfüzyon gruplarý arasýnda miyositoliz, fokal kanama ve PMNL infiltrasyonu açýsýndan fark yoktu (p>0.99). Tüm gruplar arasýnda TOD ve OSÝ kontrol grubunda en düþük, TAK ise en yüksekti. TAK, Ý/R + VA ve Ý/R + RA gruplarýnda benzer iken, bu iki grupta Ý/R grubuna göre anlamlý olarak yüksekti. Ý/R + VA grubundaki laktonaz aktivitesi kontrol grubuyla benzer iken Ý/R ve Ý/R + RA gruplarýna göre anlamlý derecede yüksekti.
SONUÇ: Çalýþmamýz vanilik ve rosmarinik asitin alt ekstremite iskemisi sonrasý kalpteki miyofibriler ödemi azalttýðýný ve TAK'ý arttýrdýðýný göstermektedir. Bununla birlikte, vanilik asit, antioksidan etkisi ile bilinen laktonaz enziminin aktivitesini rosmarinik asite göre daha fazla arttýrýr.