Olive Oil-treated Control Research Articles

Non-invasive visualization of liver fibrosis with [68Ga]Ga-DOTA-FAPI-04 PET from preclinical insights to clinical translation.

The reversibility of early liver fibrosis highlights the need for improved early detection and monitoring techniques. Fibroblast activation protein (FAP) is a promising theranostics target significantly upregulated during fibrosis. This preclinical and preliminary clinical study investigated a FAP-targeted probe, gallium-68-labeled FAP inhibitor 04 ([68Ga]Ga-DOTA-FAPI-04), for its capability to visualize liver fibrosis. The preclinical study employed [68Ga]Ga-DOTA-FAPI-04 micro-positron emission tomography (PET)/computed tomography (CT) on carbon tetrachloride-induced mice model (n = 34) and olive oil-treated control group (n = 26), followed by validation of the probe's biodistribution. Hepatic uptake was correlated with fibrosis and inflammation levels, quantified through histology and serum assays. FAP and α-smooth muscle actin expression were determined by immunohistochemistry, as well as immunofluorescence. The subsequent clinical trial enrolled 26 patients with suspected or confirmed liver fibrosis to undergo [68Ga]Ga-DOTA-FAPI-04 PET/magnetic resonance imaging or PET/CT. Key endpoints included correlating [68Ga]Ga-DOTA-FAPI-04 uptake with histological inflammation grades and fibrosis stages, and evaluating its diagnostic and differential efficacy compared to established serum markers and liver stiffness measurement (LSM). [68Ga]Ga-DOTA-FAPI-04 mean uptake in mice livers was notably higher than incontrol mice, increasing from week 6 [0.70 ± 0.11 percentage injected dose per cubic centimeter (%ID/cc)], peaking at week 10 (0.97 ± 0.15%ID/cc) and slightly reducing at week 12 (0.89 ± 0.28%ID/cc). The hepatic biodistribution and FAP expression showed a consistent trend. In the patient cohort, hepatic [68Ga]Ga-DOTA-FAPI-04 uptake presented moderate correlations with inflammation grades (r = 0.517 to 0.584, all P < 0.05) and fibrosis stages (r = 0.653 to 0.698, all P < 0.01). The average SUVmax to background ratio in theliver showed superior discriminative ability, especially between stage 0 and stage 1, outperforming LSM (area under curve 0.984 vs. 0.865). [68Ga]Ga-DOTA-FAPI-04 PET shows significant potential for non-invasive visualization and dynamic monitoring of liver fibrosis in both preclinical experiment and preliminary clinical trial, especially outperforming other common clinical indicators in the early stage. NCT04605939. Registered October 25, 2020, https://clinicaltrials.gov/study/NCT04605939.

Ameliorative effect of emblica officinalis aqueous extract on ochratoxin-induced lipid peroxidation in the kidney and liver of mice

The present study was an attempt at investigating whether the aqueous extract of Emblica officinalis may have an ameliorative effect on ochratoxin-induced lipid peroxidation in the kidney and liver of mice. Adult male albino mice were orally administered 50 microg (LD, low dose) and 100 microg (HD, high dose) of ochratoxin/0.2 ml of olive oil/animal/day for 45 days. The results revealed a significant (p < 0.05), dose-dependent increase in lipid peroxidation as well as a decreased activity of enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione transferase) and non-enzymatic antioxidants (glutathione and total ascorbic acid) in both the organs, as compared to the findings for olive oil-treated control group. Administration of Emblica officinalis aqueous extract (2 mg/animal/day) and ochratoxin for a period of 45 days caused a significant amelioration in the ochratoxin-induced lipid peroxidation in mouse liver and kidney.

Oxidized low-density lipoproteins accumulate in rat lung after experimental lung edema induced by alpha- naphthylthiourea (ANTU)

Oxidation of the low-density lipoprotein (LDL) results in the production of modified LDLs. Oxidation of LDL cholesterol plays a role on the pathogenesis of endothelial dysfunction. This study was designed to investigate the possible participation of the oxidative modification of low density lipoprotein in the lung edema induced by alpha-naphthylthiourea (ANTU), which is a well-known noxious chemical agent on the lung endothelium. When ANTU injected intraperitoneally into rats (15 mg kg −1), it produced lung edema as indicated by an increase in lung weight/body weight (LW/BW) ratio and pleural effusion (PE) reaching a maximum within 4 h. A significant lung edema was observed 4 h after intraperitoneally injection of alpha-naphthylthiourea when compared with olive oil-injected control rats. On microscopic examination of alpha-naphthylthiourea-treated rats were shown to have severe lung injury, while no change was observed in olive oil-treated control rats. While there were no staining in control lungs, positive oxidized low-density lipoproteins immune-fluorescent staining were observed in lung edema group. Our study showed that oxidized low-density lipoprotein (oxLDL) accumulated in ANTU-induced lung damage. This is the first study in which accumulation of oxLDL molecules in the intact lung tissue were shown by fluorescent immune-staining method in experimental lung edema. The potential role of oxLDL in this pathology are still under investigation.

Effect of endosulfan on ovarian compensatory hypertrophy in hemicastrated albino mice

Endosulfan, a chlorinated cyclodiene insecticide, was administered orally at 1.5, 3, 6, and 9 mg/kg per day to normal hemicastrated virgin mice for 15 consecutive days to examine the effect on ovarian function. Sham-operated and hemicastrated control mice were administered a similar volume of olive oil. The vaginal smear and body weight of the mice were recorded daily and mice were sacrificed on day 16. The remaining left ovary, uterus, kidney, adrenal, liver, thymus, and thyroid were removed and weighed. The left ovary from each animal was serially sectioned and stained for histologic studies. The hemicastrated control mice had a 40.5% increase in weight of the remaining left ovary and a significant increase in healthy and atretic follicles when compared with sham-operated controls. The remaining left ovaries of mice treated with 1.5 mg endosulfan had a 37.2% increase in weight with no significant difference in ovarian weight and or in the healthy and atretic follicles when compared with hemicastrated olive oil-treated controls. However, treatment with 3, 6, and 9 mg endosulfan resulted in weight increases in the remaining left ovary of only 14.9, 7.4, and 0.8% and a significant decrease in healthy follicles with a concomitant increase in the number of atretic follicles compared to the olive oil-treated controls. There was no significant change in the number of estrous cycles or the duration of each phase of the estrous cycle with 1.5 and 3 mg/kg per day endosulfan. However, there was a significant decrease in the number of estrous cycles, and the duration of estrus and metestrus with a concomitant significant increase in the duration of the diestrus phase with 6 and 9 mg/kg per day endosulfan treatment when compared with hemicastrated olive oil-treated controls. There were no significant change in body weight or the weights of the uterus, kidney, adrenal, liver, thymus, or thyroid after endosulfan treatment. These observations show that endosulfan treatment caused a significant decrease in compensatory ovarian hypertrophy. An increase in the number of atretic follicles and disruption of the estrous cycle may have been due to a direct effect on the ovary or to effects on the hypothalamo–hypophysial–ovarian axis.

Chlorzoxazone: a probe drug the metabolism of which can be used to monitor one-point blood sampling in the carbon tetrachloride-intoxicated rat.

In this study, we have carried out an investigation to determine if chlorzoxazone (CZX) is a suitable probe drug for predicting hepatic injury in carbon tetrachloride (CCl4)-intoxicated rats. The animals received oral doses of CCl4 (0.25, 0.5 and 1 ml/kg) 24 h prior to intraperitoneal administration of CZX. The total CYP and CYP2E1 content, as well as the aniline and CZX hydroxylase activity (Vmax and CLint), was reduced depending on the dose of CCl4 administered. At the highest concentration (128 mM) of diethyldithiocarbamate, a specific inhibitor of CYP2E1, the production of 6-hydroxychlorzoxazone (HCZX) in microsomes from CCl4-treated rats was reduced by about 85%. The IC50 value in microsomes from CCl4-treated rats was between 3 and 5 microM. The production of HCZX and the activity of aniline hydroxylase in CCl4-treated rats correlated with the amount of rat CYP2E1 protein (r=0.881, P<0.001 and r=0.822, P<0.001, respectively). The elimination of CZX by CCl4-treated rats was reduced and the HCXZ production in the CCl4-treated group was less than that in the olive oil-treated control group. The correlations between the intrinsic clearance [CLint: Vmax/Km) in vitro and the total body clearance (CLtot) of CZX hydroxylation and the elimination half-life (t1/2) of CZX in vivo in CCl4-treated rats were high (r=0.839, P<0.001 and r= -0.828, P<0.001, respectively). In addition, the metabolic plasma HCZX/CZX ratio did not require multiple blood sampling and, 2 h after CZX administration in vivo, there was also a high correlation with the CLint (Vmax/Km) in vitro (r= -0.909, P<0.01). In conclusion, these results from this study demonstrate that CZX is a good probe for monitoring the inhibition of metabolism in rats due to CCl4 treatment.

Inflammation-induced changes in serum modulate chicken macrophage function

Inflammation-induced changes in serum protein profiles and the effects of such serum on a chicken macrophage cell line HD11 were studied to find whether the changes in serum affect cellular immunity. Four-week-old male broiler chickens were injected subcutaneously with either olive oil or 50% croton oil mixed in olive oil to induce inflammation. The birds were bled at 48 h after injection, and serum protein profiles were compared using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and densitometric evaluation. At 48 h post-injection the serum from croton oil-injected birds showed distinct changes in protein profiles characterized by a selective increase or decrease in levels of several serum proteins. The protein bands which showed increases had relative molecular weights (Mr) corresponding to 65 kilo Daltons (kD), 42 kD, and two or more proteins with Mr≥200 kD. The levels of serum albumin (49 kD), and a 56 kD protein were reduced in croton oil-injected birds. The modulating effects of such serum on HD11 cells were studied using bacterial lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) induced functional activation of these cells. The LPS-induced interleukin-6 (IL-6) production by HD11 cells was not affected by the presence of either olive oil-treated control or croton oil-treated inflammatory serum but nitrite production was enhanced by the inflammatory serum. Similarly, inflammatory serum also enhanced PMA-induced respiratory burst measured using dichlorofluorescein diacetate (DCF-DA) oxidation mediated by reactive oxygen intermediates. These results suggest that inflammatory serum can modulate macrophage function by influencing the production of reactive oxygen and nitrogen species which could affect their phagocytic and bactericidal activities.

Rejection of an MHC class II negative tumor following induction of murine syngeneic graft-versus-host disease.

Cyclosporin A (CsA) has been used clinically to induce graft-versus-host disease following autologous bone marrow transplantation in an attempt to destroy residual leukemia cells and reduce relapse. To analyze the antitumor potential of murine syngeneic graft-versus-host disease (SGVHD), C3H/HeN mice were lethally irradiated, reconstituted with T cell-depleted syngeneic bone marrow (ATBM) and treated with CsA for 21 days. Graft-versus-leukemia activity was assessed by challenging groups of olive oil-treated control ATBM (OO-ATBM) and CsA-treated (CsA-ATBM) mice 1 week after CsA therapy with graded doses of the syngeneic 38C13 B cell lymphoma. Following CsA treatment, up to 70% of CsA-ATBM developed SGVHD and more than 70% of the animals injected with 500 38C13 cells exhibited long-term survival (MST >80 days). In contrast, none of the OO-ATBM control mice developed SGVHD, and more than 75% of these mice died following injection of 500 38C13 tumor cells (MST = 34 days). Long-term survivors were not resistant to tumor challenge suggesting that tumor-specific immunity did not develop. Finally, class II negative 38C13 cells cultured in IL-4 or IL-10 were not inducible for MHC class II molecules, demonstrating that class II-independent antitumor mechanisms exist in SGVHD mice.

Increased platelet-derived growth factor in the kidneys of cyclosporin-treated rats

We have examined the histological changes and the distribution of immunoreactive platelet-derived growth factor (PDGF) in the kidneys of Sprague-Dawley rats injected with cyclosporin A (CsA, 25 mg/kg/day). Control rats were injected with the olive oil vehicle alone. Groups of rats were killed after 1, 2, 3, 4 weeks of injection and 8 weeks after a 4 week period of injection. Additional controls included groups of rats injected with different doses of CsA and a group of rats subjected to chronic renal ischemia by partial of the aorta. CsA-treated rats gained weight more slowly than olive oil-treated controls (45%, P < 0.05). In rats injected with CsA, a significant increase in serum creatinine concentration (64 +/- 2 mumol/liter vs. 39 +/- 1 mumol/liter, P < 0.01) and a reduction in creatinine clearance rates (0.23 +/- 0.07 ml/min/100 g body wt vs. 0.43 +/- 0.07 ml/min/100 g body wt, P < 0.01) occurred after 3 weeks. After 1 week of CsA treatment, segments of the walls of some afferent and intralobular arterioles were thickened and stained strongly by the periodic acid Schiff (PAS) procedure. Immunostainable PDGF-BB and PDGF-AB were detected within short segments of the afferent and intralobular arterioles of the kidneys of CsA-treated rats and in the kidneys subjected to renal ischemia but not in tissue from other control animals. No PAS staining or immunostaining was evident in CsA treated kidneys eight weeks after the discontinuation of treatment. We conclude that CsA-induced ischemia results in increased accumulation of PDGF in the walls of renal arterioles.

Induction of micronuclei by vinyl acetate in mouse bone marrow cells and cultured human lymphocytes

A dose-dependent increase in micronucleated polychromatic erythrocytes was observed in the bone marrow of male C57B1/6 mice 30 h after a single intraperitoneal injection of vinyl acetate (250, 500, 1000 or 2000 mg/kg b.wt.; (9–14 animals per group). The effect was statistically significant at 1000 mg/kg (1.33±0.29 vs. 0.6±0.0% in olive oil-treated controls) and at 2000 mg/kg (1.57±0.19%) of vinyl acetate. These doses were fatal to 6 (100 mg/kg) and 8 (2000 mg/kg) out of 14 animals in both groups. The ratio of polychromatic to normochromatic cells decreased as a function of vinyl acetate dose. Cyclophosphamide (20 mg/kg), used as a positive control chemical, induced a clear increase in micronucleated polychromatic erythrocytes (2.07±0.20%). None of the treatments affected the number of micronuclei in normochromatic erythrocytes. In human whole-blood lymphocyte cultures, micronucleus induction by a 48-h treatment with vinyl acetate (0.125, 0.25, 0.5, 1 and mM; 24 h after culture initiation) was studied in lymphocytes with preserved cytoplasm from smear prepared by a method involving the removal of erythrocytes at harvest by sodium cyanide treatment to improve preparation quality. The frequency of micronucleated lymptocytes reached a peak at 0.5 mM (3.2±1.0% vs. 0.9±0.1% in control cultures) and 1 mM (3.1±0.7%), with a decline at 2 mM probably because of a toxic effect resulting in mitotic inhibition.

Effects of chlordane pretreatment on the hepatotoxicity of carbon tetrachloride

Male albine rats weighing approximately 200 g received intraperitoneal injections of chlordane (25 mg/kg) in olive oil on each of three successive days. Controls included animals given only olive oil and untreated rats. Twenty-four hours after the last dose, augmented hepatic drug-metabolizing enzyme activity in chlordane-treated rats was reflected in vivo by a reduction in zoxazolamine-paralysis time and in vitro by an increased hepatic microsomal cytochrome P-450 level. The insecticide-treated animals did not, however, display any increase of hepatic microsomal NADPH-cytochrome c reductase activity. In chlordane-treated rats, electron microscopy revealed overt proliferation of smooth endoplasmic reticulum in the hepatocytes, particularly in those located in the central one-third to one-half of the liver lobules. Olive oil-treated controls showed no alterations in paralysis time, microsomal enzyme activity, or hepatocellular ultrastructure, when compared to the untreated animals. Identically prepared chlordane-treated and control rats then were challenged with an intraperitoneal injection of carbon tetrachloride (0.5 ml of a 25% solution of CCl 4 in olive oil). Some animals from each group were killed at 4 hr after the toxic challenge; and it was determined that, in each category, there was a sharp drop in hepatic microsomal cytochrome P-450 but no change in NADPH-cytochrome c reductase, as compared to prechallenge levels. The reduction in cytochrome P-450 was most striking in the insecticide-stimulated rats. The remaining animals from each CCl 4-injected group were sacrificed at 48 hr after the toxic challenge. Histologic slides prepared from their livers revealed more extensive hepatocellular necrosis in the chlordane-pretreated rats than was found in either the olive oil-pretreated rats or the animals with no treatment prior to CCl 4 administration. It was concluded that chlordane can induce smooth-membrane proliferation and can enhance drug-metabolizing enzyme systems in rat liver and that these changes are associated with an enhanced hepatotoxic response to CCl 4 administration. It was suggested that a sharp fall in hepatic microsomal cytochrome P-450 might serve as a relatively early indicator of toxic injury in an induced liver.