Görkem EKEBAŞ, Ayhan ATASEVER, Ahmet ALPAY, Duygu YAMAN GRAM

Ratlarda karbon tetraklorür ile oluşturulan kronik karaciğer hasarı üzerine biberiye ekstraktının (Rosmarinus officinalis) etkisi ve hepatik apoptozisin belirlenmesi

Amaç: Çalışmada, karbon tetraklorür (CCl4) ile kronik karaciğer hasarı oluşturulan ratlarda Biberiye (BE) ekstraktının koruyucu etkisi, hücresel apoptozun kaspaz 3, 8 ve 9 antikorları ile değerlendirilmesi ve serum ALT (alanin amino transferaz) aktivitesi, Trigliserit, Total protein, Albümin, total Kolesterol, karaciğer MDA (malondialdehit) düzeyleri ile Total antioksidan kapasitesi (TAK) belirlendi.Gereç ve Yöntem: Çalışmada, 40 adet (200-250 gr) Wistar albino erkek rat, 10’arlı 4 gruba ayrıldı. İlk grup kontrol olup % 0,9 NaCl (0,2 mL/kg/canlı ağırlık) uygulandı. İkinci gruba 12 hafta her gün sırası ile 200 mg/kg/canlı ağırlık dozunda biberiye ekstraktı gavajla verildi. Üçüncü gruba 8 hafta boyunca haftada 2 kez 1:1 oranında mısır yağı ile dilüe edilen CCl4 (İP) 0,2 mL/kg/canlı ağırlık dozun da enjekte edildi. Dördüncü gruba 12 hafta her gün 200 mg/kg/canlı ağırlık dozunda biberiye ekstraktı gavajla verilirken aynı gruba beşinci hafta dahil 8 hafta intraperitoneal (İP) 0,2 mL/kg/canlı ağırlık olarak haftada iki doz CCl4 uygulandı.Bulgular: Grup I ve II’de karaciğer dokusu normal yapıda; Grup III’de hepatositlerde yoğun makro-mikroveziküler yağlanma, parankimde çoğunluğu lenfositlerden oluşan mononüklear hücre infiltrasyonları, nekroz, portal hafif fibrozis ile parankimde pseudolobulasyon görüldü. Grup IV’de bulgular da kısmi azalmaya rağmen Grup III’e benzer histoloji dikkati çekti. Grup I ve II’de kaspaz 3, 9 pozitif 8 negatifti. Grup III ve VI’de kaspaz 3, 8 ve 9 pozitifti. Grup IV’de kaspaz-3 gibi kaspaz-9’unda ve kısmen azalmış olması, biberiye ekstraktının sınırlı da olsa karaciğer dokusunu koruyabileceğini düşündürmüştür. Grup III’de artmış serum ALT aktivitesi Grup IV’de normal değerlere yaklaştı. Grup III’de ki serum trigliserit düzeylerindeki artışlar (P=0,001), Grup IV’de sayısal olarak azaldı. Grup III’de MDA seviyesi yüksek (P<0,05), TAK seviyesi düşükken, Grup IV’de MDA seviyesi önemli düzeyde düşüş (P<0,001). Sonuç: Grup IV’de biyokimyasal veriler karaciğer hasarının iyileştiği gösterirken, histopatolojik lezyonlar üzerine olumlu bir etki gözlenmemiştir. Bu nedenle biberiye ekstresinin farklı sürelerde ve yan etki oluşturmaksızın en iyi sonucu verecek yüksek dozlarda kullanılarak, etkisinin belirlenmesine yönelik yeni araştırmalara ihtiyaç olduğu kanaatine varıldı.

Anahtar Kelimeler:

Biberiye ekstresi, Histopatoloji, İmmunhistokimya, Karbon tetraklorür, Rat

Effect of rosemary extract on chronic liver damage induced by carbon tetrachloride in rats and determination of hepatic apoptosis

Purpose: The present study determined the possible protective effects of rosemary extract on lesions and cellular apoptosis detection by immunohistochemical method (with caspase 3, 8 and 9 antibodies) in liver tissue and some biochemical parameters; serum ALT (alanin aminotransferase) activity, triglyceride, total protein, albumin and cholesterol activity, and liver MDA (malondialdehyde) levels along with total antioxidant capacity on chronic liver injury induced by carbon tetrachloride (CCl4) in rats.Materials and Methods: Fourty Wistar albino rats (200-250 g) were divided into four groups (n:10/group). In the first group of control rats (control group), the rats were administered with %0,9 NaCl (0.2 mL/kg). The second group was administered daily with 200 mg/kg rosemary extract for twelve weeks through gastric gavage, respectively. Third group was also given CCl4 (which was diluted with corn oil, 1:1) i.p. (0,2 mL/kg) twice for 8 weeks. The rats in the fourth group rats also received 200 mg/kg rosemary extract daily for twelve weeks through gastric gavage and also given CCl4 (which was diluted with corn oil, 1:1) i.p. (0,2 mL/kg) twice for 8 weeks, starting from the 5th week.Results: The livers of the rats in group I and II had normal architecture, while an intensive macro and microvesicular steatosis, mononuclear inflammatory cell infiltrations mostly lymphocytes in portal area and paranchyma with necrotic alterations, mild to severe fibrosis with lobulation formation were observed in the group III. In Group IV, histological findings similar to Group III were noted despite partial reduction. In the group I and II, while caspase-3 and caspase-9 were positive, caspase-8 was negative. Caspase 3, caspase 8 and caspase 9 were positive in the group III and IV. There was a limited reduction in caspase 3 activity in group IV. Similar to caspase 3, partial reduction observed in caspase 9 activity suggest that rosemary extract administrations may have a partial protective effect on the liver tissue. Increased serum ALT activity level due to group IIIadministration was close to normal in group IV. Increased serum triglyceride levels in Group III were decreased numerically in Group IV. In Group III, the MDA levels were high and TAC levels were low. Compared to controls, serum triglyceride levels in CCl4 groups were determined to be higher (P=0.001). Upon the administration of rosemary extract, this value came down to lower numbers. MDA levels in in CCl4 group were higher than those of the other groups (P<0.05). The. administration of rosemary extract to CCl4 group reduced MDA levels significantly (P<0.001). There was no effect of rosemary extract administration on total antioxidant capacity. In Group IV, MDA level decreased significantly. Conclusion: Biochemical data have shown that liver damage improved in Group IV and no positive effect on histopathological lesions was observed. Since biochemical parameter values were close to normal values in groups which received rosemary extract compounds may have healing effects and therefore, new research strategies and approaches where both compounds may be tried with different periods in high doses without side effects are needed to evaluate their effects.

Keywords:

Carbon tetrachloride, Histopathology, Immunohistochemistry, Rat, Rosemary extract,

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