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

Year 2019, Volume: 28 Issue: 3, 110 - 117, 30.12.2019

Görkem Ekebaş , Ayhan Atasever , Ahmet Alpay Duygu Yaman Gram

https://doi.org/10.34108/eujhs.558310

Abstract

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.

Keywords

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

Abstract

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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