Serum GOT Levels Research Articles

Combined quercetin and simvastatin attenuate hepatic fibrosis in rats by modulating SphK1/NLRP3 pathways

Liver fibrosis involves several signalling pathways working in concert regulating the deposition of extracellular matrix. In this study, we evaluated the effect of quercetin and simvastatin alone and their combination on the treatment of experimentally induced hepatic fibrosis in rats. To decipher the potential mechanisms involved, liver fibrosis was induced in rats by administration of 40 % carbon tetrachloride (CCl4) (1 μl/g rat, i.p., twice weekly) for 6 weeks. Quercetin (50 mg/kg, orally), simvastatin (40 mg/kg, orally) either individually or combined were administered for another 4 weeks. The three treatment groups ameliorated hepatic dysfunction and altered parameters of sphingolipid and pyroptosis pathways. Yet, the combined group showed a more pronounced effect. Treatments lowered serum levels of GOT, GPT, ALP and elevated albumin and total protein levels. Histopathological and electron microscope examination of liver tissue revealed diminished fibrosis and inflammation. Protein expression levels of α-SMA, IL-1β, PPAR-γ, TGF-β1, caspase-1 and caspase-3 expression in liver tissues were reduced. Additionally, hepatic mRNA levels of SphK1 and NLRP3 decreased after treatment. Furthermore, the three groups lowered MDA levels and elevated total antioxidant capacity, GSH and Nrf2 expression levels. Treatments downregulated sphingolipid pathway and NLRP3-mediated pyroptosis and stimulated an anti-apoptotic, anti-proliferative and antioxidant activity. This suggests that targeting the SphK1/NLRP3 pathway could be a prospective therapeutic strategy against liver fibrosis.

The Effect of Chaenomelis Fructus Extract on Acute Hepatic Injury in Rats

Objective This study was conducted to investigate the effect of Chaenomelis Fructus (CF) water extract on thioacetamide (TAA)-treated rats. Methods Rats were divided into five groups: one normal group (n=8) and four with TAA-induced hepatic injury. These treatment groups were administered distilled water (n=8); silymarin 100 mg/kg (n=8); CF 100 mg/kg (n=8); and CF 200 mg/kg (n=8). In the TAA groups, the acute liver injury was induced via IP injection (200 mg/kg), and the silymarin and CF extract were then orally administered for three days. Subsequently, serum levels of GOT, GPT, and ammonia were confirmed as well as protein expressions using liver tissue. Results In the liver injury-induced rats, CF administration reduced tissue damage and serum levels of GOT, GPT, and ammonia. In addition, CF increased the anti-oxidant proteins Nrf2, Keap1, HO-1, and catalase and significantly regulated matrix metalloproteinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2). Conclusions In this animal model of liver injury induced by TAA, CF extract is determined to have a hepatoprotective effect by increasing anti-oxidant proteins that relieve damage and by regulating the expression of matrix metalloproteinases. Keywords: Chaenomelis Fructus, acute hepatic injury, thioacetamide, anti-oxidant

발효 지충이가 고지방식이를 급여한 흰쥐의 혈청 내 지질 함량 및 혈소판 응집에 미치는 영향

We investigated the effects of fermented Sargassum thunbergii (FST) on platelet aggregation and serum lipid levels in rats made obese by a high fat diet. Six-week-old male SD-rats were randomly assigned to four groups as CON, HF-CON induced by high fat diet (HF), ST supplemented with HF (HFST100), and the fermented ST supplemented with HF group (HF-FST100). After 6 weeks, the results showed that the final weight and weight gain had decreased in the HF- FST100 group compared to the HF-CON group. Also, the food efficiency ratio was significantly reduced in the HF-FST100 group compared to HF-CON. The organ weights other than heart and spleen were significantly lower in the HF-FST100 group than in the HF-CON group. The levels of serum GOT and GPT significantly decreased in the HF-FST100 group over the HF-CON group. In addition, the total cholesterol, triglyceride and LDL-cholesterol levels were lower in the HF-FST100 group than in HF-CON, while the HDL-cholesterol level was higher in the HF-FST100. The ability of platelet aggregation of groups supplemented with FST was lower than the HF-CON group. These results suggest that FST may be beneficial in improving lipid profile and platelet aggregation in obesity.

ASSOCIATION BETWEEN A31GSNP OF THE MYOSTATIN GENE AND SERUM LEVELS OF ALP, GOT, P AND CA IN EGYPTIAN BUFFALO (BUBALUS BUBALIS)

Myostatin, a member of the transforming growth factor s (TGF-s) superfamily, is a negative regulator of skeletal muscle growth in mammals. The loss or inhibition of myostatin signaling dramatically increases muscle mass. This study is aimed to detect single nucleotide polymorphisms (SNPs) in the myostatin (Mstn) gene and to associate measure their effect on mineral (Ca and P) and enzymatic (GOT and ALP) serum levels in high and low meat producing Egyptian buffaloes. Mstn gene was amplified by PCR and subsequently, subjected to sequence analysis to identify different allelic patterns. The nucleotide sequence analyses revealed presence of a novel A31GSNP in Mstn1 locus at nucleotide number 50 before exon2. This SNP was synonymous as it is located in the non coding sequence and so no amino acid was changed. However, the sequences of the other three loci showed 100% identity among the Egyptian buffaloes and as compare to the published sequences of buffalo (GenBank accession number, DQ091762). To identify whether A31G SNP is accompanied by change in serum levels of GOT, ALP enzymes, and Ca and Ph, the levels of these enzymes and minerals were estimated in the serum of high and low meat producing Egyptian buffaloes at age of 2 year. The serum levels of ALP, GOT, P, and Ca were significantly increased in high meat producing buffaloes as compared to low meat producing animals. The results of this study can be enable breeders to do an accurate and earlier selection of high meat producing buffaloes on a genotype basis.

Consumption of Seafood, Serum Liver Enzymes, and Blood Levels of PFOS and PFOA in the Japanese Population

Perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) have been shown to accumulate in the human body. The purpose of the present study was to examine the factors associated with the blood levels of PFOS and PFOA. A cross-sectional study was performed on 307 men and 301 women (aged 16-76 years) living in 15 prefectures in Japan. Blood levels of PFOS and PFOA were measured by liquid chromatography-mass spectrometry. Hepatic enzymes (γ-GTP, GOT, and GPT) and ω-3 polyunsaturated fatty acids (DHA and EPA) levels in serum were also measured. Associations between the levels of PFOS and PFOA in blood and the intake frequency of 41 kinds of dishes, foods and beverages and the serum levels of liver enzymes and ω-3 polyunsaturated fatty acids were examined using rank correlations. Frequency of intake of boiled fish in broth, sliced raw fish and coastal fish showed significant positive correlations with PFOS concentrations in blood after adjustments for potential confounders. Serum levels of GOT, GPT, DHA and EPA showed significant positive correlations with PFOS and PFOA in blood. There was also a significant regional difference in the blood levels of PFOS and 2013PFOA, with medians being highest in the Tokai/Hokuriku/Kinki region. These findings suggest that the concentrations of PFOS in blood were mainly associated with fish consumption and that the levels of PFOS and PFOA were associated with the serum levels of liver enzymes in Japanese populations. Further investigations are required to clarify the reason for the regional differences in blood levels of PFOS and PFOA in Japan.

Maternal consumption of soy protein isolate alters lipid metabolism of adult rat offspring fed ethanol diet

Previously, we observed that maternal consumption of soy protein isolate (SPI) affects lipid metabolism and cell proliferation of liver in three‐week‐old male rat offspring. Therefore, we investigated whether the adaptive responses of the offspring to environmental factors potentially affect the development of liver disease later in life. In the present study, female Sprague Dawley rats were fed a casein diet (CAS: 200 g/kg diet) or a low‐isoflavone SPI diet (SPI: 200 g/kg) for two weeks before mating, and during pregnancy and lactation. Male offspring was fed a regular chow diet until 8 weeks of age, and then pairs of offspring were fed control or alcohol liquid diet for 6 weeks. Ethanol ingestion significantly increased the relative liver weight and reduced the relative adipose tissue weight in both CAS and SPI groups. Serum GOT levels and hepatic SAM/SAH ratio were also significantly changed in offspring fed ethanol liquid diet. We observed significant interaction of maternal diet and ethanol feeding in serum triglyceride and total cholesterol levels. Maternal consumption of SPI reduced serum triglyceride and total cholesterol levels in offspring fed ethanol diet. Our data suggest that maternal consumption of SPI may program the offspring's susceptibility to the later development of alcoholic liver disease by altering lipid metabolism. This work was supported by a grant from Korea Research Foundation.

Hepatoprotective and antioxidant effects of Bridelia retusa against carbon tetrachloride-induced hepatotoxicity

The aim of present study was to validate hepatoprotective and antioxidant activities of the bark of Bridelia retusa. The aqueous ethanol extract of B. retusa exhibited highest in vitro hepatoprotective effects as evident from the significantly reduced serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) into the incubation medium of rat hepatocytes with carbon tetrachloride (CCl4), over the other organic extracts (chloroform, ethylacetate, and methanol). CCl4 administered through subcutaneous injection produced a marked elevation in the serum levels of GOT, GPT, lactate dehydrogenase, alkaline phosphatase, bilirubin, thiobarbituric acid reactive substances, and decreased in the levels of reduced glutathione, superoxide dismutase, catalase, glutathione-S-transferase, glutathione reductase, glutathione peroxidase, and total protein content. The biochemical activities were normalized in the pretreatment of rats induced by CCl4 with different doses (250 and 500 mg kg−1) of the aqueous ethanol extract of B. retusa. Histopathological changes induced by CCl4 were also significantly attenuated by aqueous ethanol extract of B. retusa treatment. The activity of the aqueous ethanol extract of B. retusa at the dose of 500 mg kg−1 was comparable to the standard drug, silymarin (25 mg kg−1). The overall data indicated that B. retusa possesses a potent protective effect against CCl4-induced hepatic damage and oxidative stress.

Manganese Supplementation Reduces the Blood Cholesterol Levels in Ca-Deficient Ovariectomized Rats

Manganese (Mn) is an essential element for normal development and bodily functions in humans. In the present study, we examined whether Mn supplementation can alter the serum lipid parameters and liver function in Ca-deficient ovariectomized (OVX) rats. Sixty female Sprague-Dawley rats (6weeks) were divided into five groups and bred for 12weeks: sham-operated control group (Sham), OVX Ca deficiency group (OLCa) with Ca-deficient diet (0.1% Ca modified AIN-93N diet), OVX Ca deficiency and Mn supplementation group (OLCaMn), OVX with adequate Ca group (OACa; 0.5% Ca AIN-93N diet), and OVX with adequate Ca and Mn supplementation group (OACaMn). A low Ca diet increased the liver weight and serum levels of GOT, GPT, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol in OVX rats. Mn supplementation decreased these parameters in Ca-deficient OVX rat. The results of our study suggest Mn supplementation results in reductions of the blood cholesterol levels, which show an increase due to Ca deficiency in OVX rats.

Hepatoprotective and Antioxidant Effects of Stereospermum suaveolens on Carbon Tetrachloride-Induced Hepatic Damage in Rats

The ethanol extract of Stereospermum suaveolens (EESS) bark was evaluated for its hepatoprotective and antioxidant activities against carbon tetrachloride (CCl4)-induced liver damage, in wistar albino rats. The ethanol extract of Sterespermum suaveolens (EESS) bark (200 and 400 mg/kg body weight, p.o.) was administered to the experimental rats for 14 days. Silymarin (50 mg/kg body weight (b.w.) was given as the standard drug. The hepatoprotective activity was assessed using various serum biochemical parameters as glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), bilirubin, and total proteins. Malondialdehyde (MDA) level as well as the activities of superoxide dismutase (SOD), reduced glutathione (GSH) and catalase (CAT) were determined to explain the possible mechanism of activity. The hepatoprotective activity was also supported by histopathological studies of liver tissue. The substantially elevated levels of serum GOT, GPT, ALP and total bilirubin, due to CCl4 treatment, were restored towards near normal by EESS, in a dose dependent manner. EESS also increased the serum total proteins of CCl4-intoxicated rats. Reduced enzymatic and non-enzymatic antioxidant levels and elevated lipid peroxide levels were restored towards near normal, by administration of EESS. A histological study showed a reduction of fatty degeneration and liver necrosis in EESS-treated rats. The results revealed that, the ethanol extract of Sterespermum suaveolens afforded significant dose dependent hepatoprotective and antioxidant effects in CCl4-induced hepatic damage.

The Effectiveness of Ethanolic Extract ofAmaranthus tricolorL.: A Natural Hepatoprotective Agent

The ethanolic extract of Amaranthus tricolor L. (ATE) leaves was tested for its efficacy against CCl₄-induced liver toxicity in rats. The hepatoprotective activity of ATE was evaluated via measuring various liver toxicity parameters, the lipid profile, and a histopathological evaluation. A sleeping time determination study and an acute toxicity test were performed in mice. The results clearly showed that oral administration of ATE for three weeks significantly reduced the elevated levels of serum GOT, GPT, GGT, ALP, bilirubin, cholesterol, LDL, VLDL, TG, and MDA induced by CCl₄. Moreover, ATE treatment was also found to significantly increase the activities of NP-SH and TP in liver tissue. These biochemical findings have been supported by the evaluation of the liver histopathology in rats. The prolongation of narcolepsy induced by pentobarbital was shortened significantly by the extract. The acute toxicity test showed that no morbidity or mortality was caused by the extract. The observed hepatoprotective effect appears to be due to the antioxidant properties of A. tricolor, which may pave the way to finding a new drug to be used for fighting liver diseases.

The Effects of Cyclooxygenase (COX)-2 Inhibition on Ischemia-Reperfusion Injury in Liver Transplantation

Purpose: Our objective was to evaluate whether COX-2 inhibition with FK3311, a selective cyclooxygenase (COX)-2 inhibitor, improves transplanted liver function. Methods: Inbred male Lewis rats weighing 200–260 g were used. The donor liver was perfused with cold University of Wisconsin (UW) solution and then stored in the same solution at 4°C for 18 hr. After the preservation period, orthotopic liver transplantation was performed. Animals were divided into three groups: the control group; the FK low-dose group (1 mg/kg FK3311 i.v. 20 min before reperfusion); and the FK high-dose group (3 mg/kg FK3311 i.v. 20 min before reperfusion). Survival rate, serum GOT and GPT levels, liver tissue blood flow, and serum thromboxane B2 (TxB2) levels were compared among groups. Results: Survival rate was significantly better (p <. 05) and serum GOT levels 30 min after reperfusion were significantly lower (p <. 05) in the FK high-dose group compared to the other two groups. Four hours after reperfusion, GPT levels and liver tissue flow were significantly (p <. 05) better in the FK high-dose group compared to the control. Both 30 min and 4 hr after reperfusion, serum TxB2 levels were significantly lower in the FK high-dose group compared to the control (p <. 05). Conclusion: COX-2 activity results in deteriorated liver function after I/R injury associated with transplantation, and selective COX-2 inhibition improved liver graft function.

Hepatoprotective and antioxidant effects of Argyreia speciosa in rats.

The present study has been designed to evaluate the liver protective and in-vivo antioxidant role of Ethanolic extract (EtAS) and Ethyl acetate extract (EAAS) of roots of Argyreia speciosa, an important 'rasayana' herb in Indian System of medicine, in CCl(4)-induced hepatotoxicity and oxidative stress in rats. Animals were treated with EtAS and EAAS at doses of 200 mg and 400 mg/kg body weight p.o. along with CCl(4) (0.7 ml/kg in olive oil, 1:1 v/v i.p. on every alternate days) for seven days. Serum biochemical parameters such as SGOT, SGPT, ALP, cholesterol, total and direct bilirubin were determined. Antioxidant status in liver was determined by measuring the activities of Super oxide dismutase (SOD), catalase and peroxidase. Histopathological study of isolated liver specimens was also carried out to know the protection offered by the extracts. There was a significant rise in the levels of serum GOT, GPT, and ALP and other biochemical parameters, decrease in the levels of SOD, catalase and peroxidase after administration of CCl(4). Suspensions of EtAS and EAAS (200 and 400 mg/kg) successfully prevented the alterations of these effects in rats (p< 0.001). Histopathological examination demonstrated that CCl(4) treated group induces ballooning degeneration and centrilobular necrosis. Groups treated with EtAS and EAAS showed recovery on ballooning degeneration and centrolobular bridging necrosis was occasionally present. Data also showed that these extracts possessed strong antioxidant activity, and were comparable to Silymarin, a well known liver protecting herbal formulation.

Evidence of gastro-intestinal system as an active and toxic target of sasanqua saponins extract

Sasanqua saponins extract (SSE) was prepared from the remainder of seeds of Camellia sasanqua after oil extract using ethanol as solvent. We have now evaluated the acute and subchronic toxicity of SSE by oral route in mice. A single oral administration of SSE (125–2000 mg/kg) produced a dose-dependent increase in adverse effects and the mortality rate of mice; the LD 50 of acute oral dose was determined as 1143.7 mg/kg for both sexes, with 95% confidence limit of 608.2–2150.6 mg/kg. In the subchronic toxicological study, SSE was administered orally at daily doses of 100, 200 and 400 mg/kg for 6 weeks. Mice of the 100 and 200 mg/kg SSE groups did not show obvious changes in the body weight, organ index and biochemical parameters, with the exception of a slight rise in serum GOT level and a decrease in triglyceride level. But mice of the 400 mg/kg SSE group died during the treated period and showed severe gastro-intestinal tract distension and submucosal changes in the small intestine. We studied further the effects of SSE on the gastric and intestinal tract function at the doses of 100 and 200 mg/kg, and found that, in mice after oral or intraperitoneal administration of SSE, the gastric empty and the small intestinal motility significantly decreased. The excretion time of feces was significantly prolonged by the oral SSE treatment. These results indicate that SSE is a kind of extract of a narrow active window and the its active and toxic target could be gastro-intestinal system.

다시마 추출물이 난소를 절제한 흰 쥐의 혈 중 지질 함량에 미치는 영향

폐경 후 여성의 비만은 독특한 생리적 현상으로 음식물 섭취가 촉진되고, 몸무게와 지방조직이 증가되고, 정상 여성보다 혈류 관련 질환 발병이 높아지는 것으로 알려져 있다. 이에 본 연구에서는 갱년기를 유도한 흰쥐에 다시마 추출물을 투여하여 체내 지질 함량 변화에 대해 그 효과를 검토하였다. 그 결과 난소를 절제한 OVX-control군이 난소를 절제하지 않은 Sham군보다 체중이 뚜렷이 증가하는 경향을 보였으며, 다시마 추출물이 체중감소에는 크게 영향을 미치지 않았다. GOT 활성에서는 난소를 절제한 OVX-control군이 난소를 절제하지 않은 Sham군에 비해 GOT 활성이 증가하였으나, 난소를 절제한 후 다시마 추출물 투여에 의해 난소를 절제하지 않은 Sham군보다 GOT활성이 더 감소하는 결과가 나타났다. GPT 활성에서는 난소를 절제 한 후 다시마 추출물을 투여한 군에서는 OVX-control군에 비해 감소하는 경향을 보였는데, 특히 OVX-ST200군에서 감소하는 결과가 나타났다. 난소 절제 후 다시마 추출물 투여는 난소 절제로 인해 야기되는 혈중 총 콜레스테롤의 함량 및 혈청 지질 농도의 증가를 감소시켰다. 또한, HDL-콜레스테롤은 난소절제 군에 비해 난소절제 후 다시마 추출물을 투여하였을 때 혈중 함량이 증가하였다. 이상의 결과에서 다시마 추출물이 갱년기 장애 시 발생할 수 있는 혈 중 지질 함량 변화에 효과가 있는 것으로 나타나 혈류 관련 질환에 대한 개선 식품으로의 활용이 기대된다. This study was performed to investigate the effects of sea tangle (ST) extracts on serum lipid concentration in ovariectomized rats. Weight-matched female Sprague-Dawley strain rats were assigned to four groups. Three groups were surgically ovariectomized (OVX). The fourth group was sham operated. Rats were randomly assigned to the following groups: sham-operated rats (Sham), ovariectomized control rats (OVX-control), ovariectomized rats supplemented with ST at 50 mg/kg body wt (OVX-ST50) and ovariectomized rats supplemented with ST at 200 mg/kg body wt (OVX-ST200). The activities of serum GOT and GPT were increased by ovariectomized. The serum GOT levels in OVX-ST50 and OVX-ST200 groups were decreased than OVX-control group. But GPT level was only decreased in the serum of OVX-ST200 group. Serum total cholesterol and triglyceride contents decreased in sham group compared with OVX-control group by ovariectomy. Six weeks feeding of ST extract resulted in a significant lowering of serum triglyceride and lowering tendency of total cholesterol Level. The serum HDL-cholesterol was higher in ST groups than OVX-control group. These results suggest that the beneficial effect of ST extract may be used to possibly improve on the lipid metabolic syndrome of menopausal women.

Inchin-ko-to prevents medium-term liver fibrosis in postoperative biliary atresia patients

The oriental herbal formulation inchin-ko-to (ICKT) inhibits liver cell apoptosis induced by transforming growth factor-beta 1 (TGF-beta1). This study evaluated the effect of ICKT on serum markers of liver function and liver fibrosis in postoperative biliary atresia (BA) patients. Twenty-one postoperative BA patients with elevated GOT, GPT and gamma-GTP, but normal serum total bilirubin levels, were divided into two groups arbitrarily; an ICKT group (n = 12), and a no-ICKT group (n = 9). Serum markers of liver function [GOT, GPT, gamma-GTP, total bile acids (TBA)], and serum markers of liver fibrosis [hyaluronic acid (HA), type IV collagen (C-IV)], were measured in both groups at the beginning of the study, and at 1, and 3 years after the beginning of the study and the results compared statistically. All patients tolerated ICKT well, and there were no side effects. In the ICKT group, mean serum HA levels were significantly decreased at 1 year (P < 0.012), and at 3 years, both mean serum HA and C-IV were significantly decreased (P < 0.001 and P < 0.003, respectively). However, mean serum levels of GOT, GPT, gamma-GTP, and TBA did not change significantly following ICKT use for any length of time (all P > 0.05). Administration of ICKT in postoperative BA patients appears to lower the serum levels of markers of fibrosis in the medium-term. Whether this in fact correlates with prevention cannot be determined from this paper, but ICKT would appear to protect against liver fibrosis. Long-term studies are required to determine the exact role ICKT plays in prognosis of BA patients.

Antioxidant activity of DHC-1—a herbal formulation

DHC-1, a multiherbal formulation, was tested for its antioxidant activity in rats. DHC-1 was investigated at dose levels of 100 mg/kg, p.o. and 200 mg/kg, p.o., once daily, for 30 days in normal rats. The levels of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), lipid peroxidation, membrane bound enzymes like Ca 2+ATPase, Mg 2+ATPase, Na +K +ATPase, lipids like phospholipid, cholesterol, triglyceride and total proteins were estimated in liver, kidneys and heart. Liver glucose-6-phosphate-dehydrogenase (G-6-P-D) was also determined. The serum levels of GOT, GPT, alkaline phosphatase, lactate dehydrogenase and bilirubin were also estimated. The decrease in the serum enzymes may be due to the membrane stabilising action of DHC-1. The inhibition of lipid peroxidation and enhancement of antioxidant enzymes (SOD and CAT) along with reduced GSH by DHC-1 may be attributed to the antioxidant potential of various ingredients present in the formulation. Thus, it can be concluded that DHC-1 exhibits an antioxidant activity and could prove beneficial in the treatment of various disorders associated with the involvement of reactive oxygen species.

Excessive Dietary Conjugated Linoleic Acid Affects Hepatic Lipid Content and Muscular Fatty Acid Composition in Young Chicks

The effects of dietary conjugated linoleic acid (CLA) on lipid concentrations and fatty acid composition of various tissues were studied in young chicks. From 7 days of age, a total of 160 chicks were divided into 4 groups, placed into 4 pens per group (10 birds per pen) and fed one of four experimental diets containing 6% tallow (TO 6%), 4% tallow plus 2% CLA (TO 4%-CLA 2%), 2% tallow plus 4% CLA (TO 2%-CLA 4%) or 6% CLA (CLA 6%) for 3 weeks. There were no significant differences in growth performances and the relative weights of various organs, but relative liver weight of chicks fed dietary CLA at 4 and 6% levels was significantly higher (p<0.05) than that of TO 6% group. The chemical compositions of leg muscle were not affected by CLA feeding. However, hepatic total lipid of chicks fed 6% CLA diet was significantly higher (p<0.05) than those of TO 6% and TO 4%-CLA 2% groups. The concentrations of various lipid fractions in serum were not affected by CLA feeding. With the increase in dietary CLA levels, cis 9-trans 11 CLA, trans 10-cis 12 CLA and total CLA of leg muscle increased linearly. The relative proportions of C18:1 ω-9 and C20:4 ω-6 fatty acids in the leg muscles of chicks fed the CLA containing diets were significantly lower (p<0.05) than those of TO 6% group. These results indicate that the levels of CLA isomers were increased linearly in dose-dependent manner after feeding of synthetic CLA source. But it was also observed that excessive amount of dietary CLA resulted in the possible adversely effects, such as increase of liver weight, hepatic lipid accumulation and serum GOT level.

Transcriptional slippage of p53 gene enhanced by cellular damage in rat liver: monitoring the slippage by a yeast functional assay

The Long–Evans Cinnamon (LEC) rat is a mutant strain characterized by abnormal copper metabolism and a high incidence of hepatitis and hepatoma. Using a yeast-based assay which scores mutants in p53 gene transcripts as red colonies, we detected frequent mutations in the liver of LEC rats. The majority (50–60%) of these were frameshift mutations caused by the insertion of an extra adenine (A) in the regions containing six consecutive adenines. The rate of A insertion was calculated to be 6.9–9.0% of the total p53 cDNA. Insertions of an extra adenine were found almost exclusively in the mRNA (cDNA), especially in the (A) 6 tract located at the most 5′-side (exon 4) among the three (A) 6 tracts (exons 4, 7, and 8), but rarely in the corresponding sites of genomic DNA. Wild-type p53 cDNA was transcribed in vitro into mRNA with the use of SP6 RNA polymerase and tested by the yeast functional assay. Subsequent sequencing detected A insertions at an overall rate of 1.6% in exons 7 and 8 but none in exon 4. This indicates that the A insertion in the exon 4 (A) 6 tract was an in vivo phenomenon rather than an artifact in reverse transcription or polymerase chain reaction. The percentage of red colonies increased sharply to about 20% of the liver samples in the acute hepatitis stage, and returned to control level of those in the chronic hepatitis stage, and increased again slightly to those in the neoplastic stage. The percentage of red colonies correlated with the serum GOT level ( r=0.96, p<0.001) but not with the contents of copper and 8-hydroxydeoxyguanosine in the liver of LEC rats. Ethanol treatment of hepatic cell lines also increased the rate of transcriptional slippage at the (A) 6 tract. These findings indicate that cellular damage is responsible for the increase in the rate of mutation at the transcriptional level, and suggest that cellular damage degrades transcriptional fidelity, thereby further impairing cellular functions.

Antioxidant activity of AO-8, a herbal formulation in vitro and in vivo experimental models.

The effect of AO-8, a herbal formulation was evaluated for free radical scavenging activity using in vitro and in vivo experimental models. AO-8 dose dependently inhibited ferric ion induced lipid peroxidation in vitro at 125-1000 micrograms. AO-8 was investigated at dose levels of 250, 500 and 750 mg/kg p.o. in isoproterenol-induced myocardial infarction and CCl4-induced hepatotoxicity in rats. The levels of serum GOT and LDH, cardiac glutathione, SOD and catalase were estimated following induction of myocardial infarction with isoproterenol. The estimation of parameters such as SGOT, SGPT, liver glycogen and lipid peroxidation were carried out in CCl4-induced hepatotoxicity. Treatment with AO-8 for 15 days at a dose of 500 and 750 mg/kg offered marked protection in both in vivo models. The reversal of increased serum enzymes in both the models may be due to the prevention of leakage of the intracellular enzymes by its membrane stabilizing activity. The inhibition of lipid peroxidation and enhancement of antioxidant enzymes by AO-8 may be due to the direct free radical scavenging activity which could be attributed to the antioxidant potential of various ingredients present in the formulation. Thus it can be concluded that AO-8 is an effective free radical scavenger and could prove beneficial in the treatment of various disorders associated with the involvement of free radicals.

CD4-TARGETED THERAPY ABROGATES ACCELERATED REJECTION OF LIVER ALLOGRAFTS IN PRESENSITIZED RAT RECIPIENTS

127 Allograft (Tx) rejection in sensitized patients represents one of the most challenging problems in clinical transplantation. We have developed a model of accelerated rejection in which LEW rats presensitized with BN skin Tx (day-7) reject LBNF1 cardiac Tx (transplanted at day 0) in <36 h. We have also shown that treatment with RIB-5/2, a nondepleting CD4 mAb, may induce tolerance in this sensitized Tx model. However, to achieve tolerogenic effect, animals require intermittent infusion of CD4 mAb in the "sensitization" phase, i.e. starting at the time of skin Tx; CD4-targeted therapy after cardiac engraftment is ineffective. This study was designed to establish a model of accelerated rejection of orthotopic liver Tx (OLT) and then to test the effects of RIB-5/2 mAb treatment in sensitized rat recipients. First, we have determined that normal LEW rats survived >100 days following transplantation of LBNF1 livers (Gr. I; n=6), similarly to isograft controls (n=3). Sensitization with BN skin Tx at day-7 prompted accelerated rejection of liver Tx, and resulted in animal death within 8.9±2.2 days (Gr.II; n=11). Then, in order to mimic a clinical scenario, we asked whether CD4-targeted therapy introduced in presensitized hosts at the time of liver engraftment may affect the accelerated rejection cascade. Accordingly, LEW rats bearing BN skin Tx for a week were treated by RIB-5/2 mAb immediately after OLT (7× 5mg/rat i.v. day 0 to 21). Indeed, all animals in this treatment group are currently surviving at >50 days post-Tx and maintain good liver function (Gr. III; n=6). Hence, by day 4, serum GOT levels (IU/L) in Gr. III recipients were 140±51 vs. 348±53 and 840±51 in Gr. I and Gr. II rats, resp. (p<0.01; n=3/group). By day 50, sGOT levels in the treatment group were 233±92 IU/L (n=3); no circulating RIB-5/2 mAb were detectable at 3 weeks after cessation of RIB-5/2 mAb therapy. Liver samples were collected from recipient groups at day 8 post-Tx for histological evaluation, and the degree of rejection was scored according to the Banff criteria by blinded pathologist. Unlike in isografts, which remained essentially normal, severe lobular disarray with diffuse necrosis and fibrin deposition in hepatic sinusoids, was noted in Gr. II sensitized rats, consistent with ferocious accelerated OLT rejection. In marked contrast, Gr. III liver Tx harvested from RIB-5/2 mAb treated presensitized hosts, showed only mild bile duct obstruction and proliferation, consistent with "mild" acute OLT rejection. Liver Tx harvested at day 8 from Gr. I unsensitized hosts showed signs of "moderate-severe" acute rejection. Finally, we analyzed host systemic IgM and IgG allo-Ab responses in sera samples by flow cytometry (n=3/group/time-point). No differences in IgM and IgG allo-Ab levels could be detected at day 7 post-Tx between animal groups. Interestingly, however, by day 28, RIB-5/2 mAb treatment depressed IgM (mean channel fluorescence±SD = 28±30 vs. 376±108 in Gr. I; p<0.01), and prevented the switch from IgM to IgG (0 vs. 435±27 in Gr. I; p<0.01). In conclusion, this study documents the efficacy of a clinically relevant nondepleting CD4 mAb regimen to abrogate accelerated rejection and induce long-term acceptance of liver Tx in sensitized rat recipients. Mab-facilitated depression of host cellular and humoral immunity may contribute to OLT survival in this stringent transplantation model.