Equal Volume Of Olive Oil Research Articles

24 Effects of Oral Supplementation of Cannabidiol on Young Stallion Behavior

Abstract Cannabidiol (CBD) has become a popular product in the recent months due to change in legislation with the passing of the Agricultural Improvement Act in 2018. Cannabidiol comes from the plant Cannabis sativa L. and binds to cannabinoid receptors throughout the endocannabinoid system. These receptors are located throughout the body and are prominent in the brain and reproduction tract. Although the effects of CBD on physiology have been researched, little data on the behavioral effects exist, even less so in horses. Calming supplements that may negate aggressive stallion behavior are not suitable due to potential soporific effects that can affect libido. In addition to the effectiveness of CBD to modify behavior in horses, it is also unclear if CBD influences stallion libido. The objective of this study was to evaluate sexual behavior in stallions given a daily CBD supplement for 70 days. Two-year old stallions (n = 8) were assigned to either a Control (CON; n = 3) or Cannabidiol (CBD; n = 4) treatment groups. Cannabidiol was administered at a rate of 0.6 mg/kg BW daily, and equal volume of olive oil was given as a placebo to CON. Horses were weighed every Monday at which time dosages were recalculated to adjust for body weight changes. Behavior data collection occurred twice per week during semen collections. Video data were recorded by placing two cameras, one placed in view of the tease area, and one placed in view of the mount, for behavior data backup. Behavioral data was tallied by handlers calmly calling each behavior as it occurred during collection. The behaviors collected included Flehman’s response, bites, strikes, mounts, high pitch vocalizations, and low pitch vocalizations. Data were analyzed using a Chi Square Test within R Statistical Program (R Core Team). Results were considered significant at P < 0.05. Throughout the duration of the study, all stallions exhibited normal sexual behavior, regardless of treatment. Results showed that total low and high pitch vocalizations, bites, and Flehman’s were affected by treatment (P < 0.01). Cannabidiol-treated stallions had more low pitches (60), compared to CON (58). Cannabidiol-treated stallions had fewer bites (145) compared to the CON (146). Cannibidiol stallions displayed Flehman’s more frequently (270) than CON (236). Flehman’s response and bites were lower for stallions on cannabidiol in Week 1 but increased by Week 10 (P < 0.01), while only Flehman’s response increased in the CON group over time (P < 0.01). These findings indicate that CBD may influence breeding behavior in young stallions specifically when supplemented over time. Further research should focus on the impact of CBD on breeding behaviors in mature stallions and the longevity of the behavioral response to CBD.

136 The effects of cannabidiol on libido and heart rate in stallions

Changes in the Agriculture Improvement Act of 2018 increased popularity of Cannabidiol (CBD) as a supplement. Exogenous CBD, derived from Cannabis Sativa L., binds to cannabinoid receptors in the endocannabinoid system. These receptors are located throughout the body, CB1 being prominent in the central nervous system and CB2 in the immune system. Physiological effects of CBD have been well researched but the effects on libido have not. Lack of libido or sexual drive can be detrimental to stallion breeding productivity. The use of CBD as a calming supplement in animals is increasing due to marketing strategies. The study objective was to evaluate libido and heart rate (HR) in stallions given a daily oral CBD supplement for 90d. Two-year old stallions (n = 7) were assigned to either Control (CON; n = 3) or Cannabidiol (CBD; n = 4) treatment groups. CBD was administered at 0.6mg/kg of BW/d, while CON was administered an equal volume of olive oil, as a placebo of similar nature. Treatments were top-dressed over morning feed. Horses were weighed every Monday with dosages recalculated accordingly. Data collections occurred 2x/wk during semen collections with an artificial vagina. Behaviors were previously defined, calmly called out and logged as they occurred. Observations included time spent licking a tease mare, time to erection, time to first mount, and time to ejaculation. A Polar® heart rate monitoring system linked to the KER® ClockItapp was usedto measure HR. Variations of HR included resting HR, teasing HR, mounting HR, step-off HR, and walk-off HR. Data was analyzed using a repeated measures ANOVA within R Statistical Program®(R Core Team, 2022). Results were considered significant at P ≤ 0.05. Throughout the duration all stallions exhibited normal sexual behavior, regardless of treatment. A treatment affect was not seen for licking time between CBD and CON (11.92s ± 3.76 and 0.32s ± 0.20 respectively). There was no difference between CBD and CON for time to erection (27.64s ± 6.84 and 22.9s ± 4.76 respectively). Time to first mount CBD stallions averaged 330.6s ± 23.04, CON stallions averaged 359.5s ± 25.51, with no effect of treatment. Time to ejaculation in CBD stallions was 356s ± 23.78 and 374s ± 24.84 in CON stallions. There were no differences in behaviors over time. There was a time x day effect for walk-off HR(P = 0.04), though this was significant in both groups. No other differences in HR occurred. Long-term use of CBD for young stallions was seen to not affect libido. However, supplementation may lower HR over time. The immediate and lasting response to CBD use in mature stallions and older aggressive stallions is needed to understand the effects of using these products.

Double-blind RCT of fish oil supplementation in pregnancy and lactation to improve the metabolic health in children of mothers with overweight or obesity during pregnancy: study protocol

IntroductionMaternal obesity during pregnancy is associated with adverse changes in body composition and metabolism in the offspring. We hypothesise that supplementation during pregnancy of overweight and obese women may help...

Ginsenoside Rg1 alleviates ANIT-induced intrahepatic cholestasis in rats via activating farnesoid X receptor and regulating transporters and metabolic enzymes

Ginsenoside Rg1 is an active ingredient extracted from the roots of ginsenoside, and an α-naphthylisothiocyanate (ANIT)-induced rat model of intrahepatic cholestasis was used to investigate the protective effect of Rg1 on cholestasis. 48 SD male rats were randomly divided into 6 groups: control group, model group, UDCA group (ursodeoxycholic acid), low-dose Rg1 group (10 mg/kg), medium-dose Rg1 group (20 mg/kg) and high-dose Rg1 group (40 mg/kg). The model group, the UDCA group and all the Rg1 group were then intragastrically administered with 80 mg/kg ANIT, and the control group were given equal volume of olive oil. Then the pathological changes in liver tissue were observed, the secretion of bile in the bile duct was measured, and the biochemical markers in serum were quantified, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutamyl transfer peptidase (GTP) and the content of total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA). The contents of inflammatory mediators in serum were quantified, including tumor necrosis factor (TNF-α), γ-interferon (IFN-γ) and interleukin-1β (IL-1β). The contents of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in liver homogenate were quantified. Expression of farnesoid X receptor (FXR), transporters and metabolic enzymes in liver tissue was monitored. Rg1 treatment improved liver tissue pathological damage, promoted bile secretion and significantly reduced serum levels of the intrahepatic cholestasis markers ALT, AST, ALP, GTP, TBIL, DBIL and TBA. Rg1 increased the activity of SOD and GSH-Px in liver homogenate, while, reducing the serum levels of MDA and inflammatory mediators. Rg1 also regulated the expression of FXR, bile acid transporters and metabolic enzymes. Overall, Rg1 alleviated liver injury by improving secretion of bile and normalizing the activity of enzymes in the serum. The protective mechanism appeared to be related to the activation of FXR and regulation of liver transporters and metabolic enzymes.

Down-regulation of microRNA-31 suppresses hepatic fibrosis induced by carbon tetrachloride

MicroRNA-31 (miR-31) is among the most frequently altered microRNAs in human diseases, and altered expression of miR-31 has been detected in a large variety of diseases types. miR-31 could also regulate a variety of cell functions including hepatic fibrosis. Hepatic stellate cells (HSCs) are regarded as the major cell type involved in hepatic fibrosis. Male BALB/c mice (five mice per group aged 6 weeks) received 200 μL of body weight of carbon tetrachloride (10% CCl4) mixed with olive oil intraperitoneally, and the first dose was doubled. To induce hepatic fibrosis, carbon tetrachloride was injected twice a week for 4, 6, 8, and 10 weeks. Control animals were injected with an equal volume of olive oil at the same time intervals. We found that miR-31 expression and fibrosis-related factors in four hepatic fibrosis stages. However, we noted that inhibition of miR-31 was down-regulated fibrosis-related factor expression in F1–F3 stages, but no F4 stage. Thus, we hypothesize that miR-31 may mediate hepatic fibrosis. In this research, we found that inhibition of miR-31 expression significantly inhibited HSC activation. The biological function of miR-31 during HSC activation might be through targeting hypoxia-inducible factor 1-alpha inhibitor (HIF1AN). Inhibition of miR-31 can reduce the transcription factor activity of hypoxia inducible factor 1 (HIF-1) by targeting the biological effects of HIF1AN with the condition of hypoxia. In later hepatic fibrosis could be rescue combining with inhibition of miR-31 and adding heparin-binding EGF-like growth factor (HBEGF).

Study on anti- fibrotic mechanism of Fuzheng -Huayu formula to suppress autophagy in mice

Objective: To determine whether the anti-hepatic fibrosis effect of Fuzheng-Huayu formula is related to suppress autophagy in mice. Methods: C57 mice were randomly divided into normal group (N group) and model group. The model group was induced by intraperitoneal injection of carbon tetrachloride to induce liver fibrosis in mice, and the normal group was injected with equal volume of olive oil. After 1 week, the model group was randomly divided into model (M) group, rapamycin (Rapa) group, rapamycin plus chloroquine (Rapa+CQ) group, rapamycin plus salvianolic acid B (Rapa+Sal B) group, rapamycin plus Fuzheng -Huayu formula (Rapa+FZ) group. Each drug group was administered corresponding drugs by gavage on a daily basis, and N group and M group were given the equal amount of drinking water by gavage. After 5 weeks, the mice were sacrificed, and HE and Sirius red staining were used to observe the inflammation and collagen deposition on liver tissue in each group. The hydroxyproline content was determined by alkaline hydrolysis method. Western blotting was used to detect changes in the expression of autophagy in liver tissue and microtubule-associated protein 1 light chain 3II/I (LC3II/I), p62, α-smooth muscle actin (ɑ-SMA) and type I collagen expression. Immunofluorescence staining was used to observe the immunofluorescence localization of ɑ-SMA and LC3B in liver tissues of each group. ). A t-test was used to compare the two independent samples. LSD or Dunnett's T3 test were used to compare the mean of multiple samples. Results: There was no significant difference in N and M groups in terms of body weight. The body weight of the mice in each drug group decreased significantly (F = 14.041, P < 0.001). The liver/spleen /body weight ratios of each drug group and M group were significantly higher than the N group (F = 26.992, 6.589, P < 0.001). The expression of p62 protein in the liver tissue of mice in each drug group was lower than M group, and the difference between Rapa group and Rapa+Sal B group (F = 3.085, P = 0.039, 0.003) was statistically significant, while that of Rapa + Sal B group was lower. Compared with group M, the expression of LC3B II in Rapa group was significantly higher (F = 7.514, P = 0.01). Immunofluorescence staining showed that LC3B and α-SMA CO-stained cells were absent in the liver of mice in N group, and co-stained cells were found in the liver of mice in M group. The co-stained cells in the liver of mice in each drug group were significantly higher than M group, and the co-stained cells in Rapa+FZ group were fewer. Compared with the N group, the collagen deposition of M group and each drug group was significantly increased; the collagen deposition of each drug group was lower than that of the M group. There was no statistically significant difference between each drug group. Compared with N group (77.75 + 48.79), hydroxyproline in liver tissue of mice in M group was significantly increased (293.48 + 84.43) (F = 3.015, P = 0.005), and the content of hydroxyproline in liver tissue of mice in each drug group was lower than M group, but the difference was not statistically significant (F = 0.750, P = 0.573). Compared with the N group, the expressions of α-SMA and type I collagen in the M group were significantly increased (F = 27.718, 18.893, P < 0.01). The expression of α-SMA in Rapa group and Rapa+Sal B group was similar to M group, while Rapa + CQ group and Rapa + FZ group were significantly lower than Rapa group and M group (P < 0.01). The expression of type I collagen in Rapa + CQ group was significantly higher than Rapa group (P = 0.017), while the expression of type I collagen in Rapa + FZ group was significantly lower than M group (P = 0.013). Conclusion: Autophagy of hepatic stellate cells was observed in carbon tetrachloride-induced liver fibrosis model. Rapamycin can promote autophagy in hepatocytes and hepatic stellate cells. Fuzheng-Huayu formula and Salvianolic Acid B might antagonize the effect of rapamycin on autophagy.

Hepatoprotective properties of methanol leaf extract of Pterocarpus mildbraedii Harms on carbon tetrachloride- induced hepatotoxicity in albino rats (Rattus norvegicus)

Aim: This study evaluated the effects of methanol leaf extract of Pterocarpus mildbraedii Harms on carbon tetrachloride (CCl4)-induced sub-acute hepatotoxicity in albino rats. Methods: Fresh leaves of P. mildbraedii were collected in December 2016. Thirty male albino rats were randomly assigned to 6 groups (A – F) of 5 rats each. Sub-acute hepatotoxicity was induced in groups A, B, C, D and E by intraperitoneal injection of 1ml/kg CCl4 in equal volume of olive oil at 3-day intervals for 12 days. Group A was given 10 ml/kg distilled water placebo and served as untreated (negative) control, groups B, C, and D were treated with 100, 200 and 400 mg/kg P. mildbraedii methanol leaf extract (PME) respectively, group E was treated with 100 mg/kg silymarin as positive control, while group F was given 10 ml/kg distilled water placebo and served as normal control. Treatment with PME and silymarin was done orally twice daily for 15 days. Blood samples were collected on the 15th day for evaluation of liver enzyme markers (alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities) and liver function (total serum protein, albumin, globulin, total cholesterol and bilirubin), following standard procedures. Relative liver weight was calculated. Results: Treatment with PME and silymarin significantly (P < 0.05) decreased the elevated ALT and AST, and thus restored hepatocellular integrity, and also ameliorated inflammatory liver enlargement in the rats given CCl4. Conclusion: These findings imply that treatment with PME as used in this study led to significant hepatoprotection against CCl4-induced hepatotoxicity.

Protective Role of Pomegranate peel extract on the Pituitary gland of adult male rabbits treated with CCL4

This study was carried out to investigate the protective role of pomegranate peel extract for relieving the harmful effects of carbon tetrachloride on the pituitary gland function in adult male rabbits. Twenty four adult male rabbits were used in this study and divided randomly into four equal groups. Rabbits of the 1st group were received 1 ml of orally distal water once a day and olive oil 1/2 ml /kg B.W. intraperitoneal injection [I.P] twice a week as a control group. The second group were treated I.P with 500 mg / kg B.W. of CCL4 mixing with equal volume of olive oil (o.5 ml/kg B.W.) twice a week (group T1). The third group were received orally pomegranate peel extract (100 mg/kg B.W) once a day and olive oil 1/2 ml /kg B.W. I.P twice a week ( group T2). The fourth group were received pomegranate peel extract (100 mg/kg B.W) once a day and treated I.P with 500 mg / kg B.W. of CCL4 mixing with equal volume of olive oil (o.5 ml/kg B.W.) twice a week (group T3). Blood samples were collected at on days (0, 14, 28.42 and 56) for measuring LH, FSH concentrations, total serum protein after treatment. Results showed a significant (P&lt;0.05) decrease in total serum protein in T1 group compared with control group. While T2 group showed a significant (P&lt;0.05) increase in these parameter as compared with the control and T1 group at the end of treatment period. T1 group indicated i a significant (P&lt;0.05) increase in serum LH at(zero and 14 day) and FSH at( from zero time to the 28 day) . These levels, however, were significantly decreased in T1 group compared to what we observed in the control ones at the end of the experiment period. T2 group also showed a significant (P&lt;0.05) increase in serum FSH at (24 day to the 56day of experiment) and LH concentration (at 56 day) compared to control group. Histological study was showed sever congestion of blood vessel and necrotic area in section of anterior pituitary gland in group treated with CCL4, while demonstrate moderate congestion in T3 group.&#x0D; In conclusion, Pomegranate peel extract at dose (100 mg/kg B.W.) has positive effects on the pituitary gland function, hence eliminating the deleterious effects or toxic effect of CCL4.

Effects of the Carbon Tetra Chloride on the Body Weight and Ovaries, Uterus, Testes of the Rabbit

Aims: To investigate the effects of carbon tetra chloride on the body weight and ovaries, uterus, testes of the rabbit for 30 days. Methodology: The rabbit were randomly divided into two equal groups (control and one treated groups), and the animals were treated as follow: 1- Control group C. (n = 15) received Distilled water 1ml /kg b.w four times a week intrapertunium for 30 days. 2- Treatment group. (n=15) were treated with carbon tetrachloride (1 mg / kg b.w mixing with equal volume of olive oil (1ml/ kg b.w.) four times a week/ intrapertunium for 30 days. The body weight of the animals detected at the day 1 th , 7 th , 14 th , 21 th and 30 th of the experimental period. At30 days animals sacrificed and ovaries, uterus, testis were removed and weighted, Samples of ovaries, uterus & testis were taken for histopathological study. Results: Treatment with carbon tetra chloride causes a significant decrease in body weight. Also a significant decreased of ovarian, testicular weight and non-significant increase of uterus weight's. Conclusion: Carbon tetra chloride causes decrease body weight, degeneration testes, ovaries followed by effect on fertility.

Artificial oil body as a potential oral administration system in zebrafish

Zebrafish are well recognized as an excellent vertebrate model which, in particular, facilitates the rapid investigation of drug effects on cancers. A prerequisite for this anti-cancer study appears to have a non-invasive and effecicve method for admininstration of drugs. In this study, we explored the potential of artificial oil bodies (AOBs) for the oral administration of a hydrophobic agent in adult zebrafish. A hydrophobic dye was employed for illustration, and AOBs were assembled with phospholipids, a storage protein, and an equal volume of olive oil and brine shrimp oil. Consequently, more than 90% of the dye was encapsulated in AOBs which exhibited a prolonged release profile for the dye cargo. The dye-loaded AOBs collected as an oil cake remained integral within 30 min in the aquatic solution while it was completely consumed by one zebrafish in a few seconds. As revealed by the fluorescence microscopy and frozen section, the fluorescence signal was detected in the zebrafish intestinal tract 2 h after the oral administration. The dye cargo was completely absorbed in 12 h and almost no trace was left in zebrafish feces. Overall, the AOBs system is biocompatible and shows promise in the oral administration of chemical entities for zebrafish.

Protective Role of Pomegranate Peel Extract on Testis in Adult Male Rabbits Treated with Carbon Tetrachloride

The aim of the present study is to prepare ethanol extract of Pomegranate peel and the effects of this extract on testicular weight to body weight ratio, Serum cholesterol, testosterone concentration and histopathological changes of testes in rabbits treated with carbon tetrachloride (CCl4). Twenty four adult male rabbits were used. They were divided randomly into four equal groups. Animals were treated for 56 days as following: Rabbits of the 1st group were received 1 ml distal water orally once a day and olive oil 0.5 ml /kg B.W. I.P twice a week as control group. The second group were treated I.P with 500mg / kg B.W. of CCl4 mixing with equal volume of olive oil (0.5 ml/kg B.W.) twice a week (group T1). The third group was received pomegranate peel extract orally (100 mg/kg B.W) once a day and olive oil 0.5 ml /kg B.W. I.P twice a week (group T2). The fourth group were received pomegranate peel extract (100 mg/kg B.W) once a day oral I.P with 500 mg / kg B.W. of CCl4 mixing with equal volume of olive oil (o.5 ml/kg B.W.) twice a week (group T3). Blood samples were collected at (0, 14, 28, 42 and 56) days for measuring testosterone concentration, Serum cholesterol after treatments. Animals weighed and scarified and testis were removed and weighed, Samples of testis were taken for histopathological study. The results of the present study showed that treatment with pomegranate peel extract causes a significant (P&gt;0.05) increase in testicular weight to body weight ratio. Also a significant (P&gt;0.05) decreased of serum cholesterol and a significantly (P&gt;0.05) elevation of testosterone concentration were observed. Histopathological examination of the testis was revealed that the extract of Pomegranate peel protect the testis against lesions caused by CCl4. In conclusion, Pomegranate peel extract could protect the tissue of testicles from CCl4 perhaps, by its anti-oxidative effect of pomegranate peel extract, hence eliminating the deleterious effects or toxic effect of CCl4.

DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide in implantation phase

To investigate the DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide (CS2) in the implantation phase and to explore the mechanism of abnormal implantation induced by CS2 from the perspective of immune injury. Mice were exposed to CS2 at different doses or at different time points in the implantation phase to establish model 1 and model 2. For model 1, mice were assigned to four groups to receive a single intraperitoneal injection of low-dose CS2 (0.1 LD50, 157.8 mg/kg), middle-dose CS2 (0.2 LD50, 315.7 mg/kg), and high-dose CS2 (0.4 LD50, 631.4 mg/kg) as well as an equal volume of olive oil (control) on gestational day (GD) 4. For model 2, mice were assigned to four groups to receive a single intraperitoneal injection of CS2 (0.4 LD50, 631.4 mg/kg) or an equal volume of olive oil (control) on GD3, GD4, GD5, and GD6. At the end, single cell suspension of splenic lymphocytes was prepared. Cell viability was measured by trypan blue staining, and the DNA damage of splenic lymphocytes was evaluated by alkaline single cell gel electrophoresis assay. The middle-dose and high-dose exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P < 0.01); there was significant regression relationship between indicators of DNA damage and exposure doses (P < 0.01). The GD3, GD4, GD5, and GD 6 exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P < 0.01), and the GD 4 exposure group had the most DNA damage. Exposure to CS2 in the implantation phase can induce DNA damage of splenic lymphocytes in pregnant mice, and the DNA damage was aggravated with the increase in CS2 concentration. GD4 may be the sensitive time point for DNA damage of splenic lymphocytes induced by CS2 in pregnant mice.

Effects of polybrominated diphenyl ether-153 lactation exposure on the concentrations of intracellular calcium ion and calcium-activated related enzymes levels of adult rats' cerebral cortex

To investigate the effects of polybrominated diphenyl ether-153 (BDE-153) exposure during lactation period on the calcium ion (Ca(2+)) concentration and calcium-activated enzyme levels in cerebral cortical cells among adult rats and to provide a scientific basis for the study on the developmental neurotoxicity of BDE-153. Forty newborn male rats were randomly and equally divided into four groups according to their body weights and litters: 1, 5, and 10 mg/kg BDE-153 groups and olive oil solvent control group. On postnatal day 10 (PND 10), the BDE-153 groups were administrated BDE-153 (0.1 ml/10 g body weight) by intraperitoneal injection, while the olive oil solvent control group was given an equal volume of olive oil. Two months later, these rats were decapitated, and the cerebral cortex was separated quickly on an ice-cold dish. The Ca(2+) concentration in cerebral cortical cells was measured by flow cytometry. The activities of calcineurin (CaN) and Ca(2+)-Mg(2+)-ATP enzyme were determined by colorimetric method. The mRNA and protein expression of calpain-1 and calpain-2 was measured by real-time quantitative PCR and Western blot. The mean fluorescence intensities of intracellular Ca(2+) in control group and 1, 5, and 10 mg/kg BDE-153 groups were 10.83, 1.48, 1.93, and 0.62, respectively; the 1, 5, and 10 mg/kg BDE-153 groups had significantly lower intercellular Ca(2+) concentrations than the control group (P < 0.05). The activities of CaN and Ca(2+)-Mg(2+)-ATP enzyme and mRNA and protein expression of calpain-1 showed no significant differences between the 1, 5, and 10 mg/kg BDE-153 groups and control group (P > 0.05). The protein expression of calpain-2 increased as the dose of BDE-153 rose. Compared with the control group (mRNA: 0.81±0.26; protein: 0.15±0.07), the 5 and 10 mg/kg BDE-153 groups had significantly higher mRNA expression of calpain-2 (5 mg/kg BDE-153 group: 1.16±0.52; 10 mg/kg BDE-153 group: 1.32±0.23) and significantly higher protein expression of calpain-2 (5 mg/kg BDE-153 group: 0.31±0.07; 10 mg/kg BDE-153 group: 0.37±0.06) (P < 0.05). The 10 mg/kg BDE-153 group had significantly higher protein expression of calpain-2 than the 1 mg/kg BDE-153 group (0.37±0.06 vs 0.22±0.07, P < 0.05). Ca(2+-) mediated calpain-2 activation may be one of the main mechanisms of BDE-153 neurotoxicity.

The Protective Role of Date Palm Pollen (Phoenix dactylifera L.) on Liver Function in Adult Male Rats Treated with Carbon Tetrachloride

The present study was carried out to investigate the protective role of date palm pollenaqueous suspension against the toxic effects of carbon tetrachlorideon liver function in adultmale rats by studying the following parameters,estimation of ALP, AST, ALT enzymesactivity, totalserum bilirubin, and histological study of liver.Forty adult male rats aged 12-14weeks and weighed 275-325 gm were randomly divided into four equal groups(10 rats/group)and were treated for 42 days as follows : rats of the first group were received 1 ml tap waterorally once a day and olive oil 0.5 ml /kg B.W. intraperitonelly twice a week whichconsidered as group C, rats of the second group were received date palm pollen suspensionorally (150 mg/kg B.W) once a day and olive oil 0.5 ml /kg B.W. intraperitonelly twice aweek( group T1),rats of the third group were treated intraperitonelly with 500mg / kg B.W. ofCCL4 mixed with equal volume of olive oil twice a week (group T2) ,while rats of the fourthgroup were received date palm pollen suspension (150 mg/kgB.W) once a day orally andtreated intraperitonelly with 500 mg / kg B.W. of CCL4 mixing with equal volume of oliveoil twice a week (group T3). The blood samples were collected at (zero, 14, 28, 42) days forbiochemical parameters. After that, six rats from each group were sacrificed, and thensamples of liver were taken for histological study. The results revealed no significantdifferences (P &gt; 0.05)in Liver Enzymes activity (ALT , AST , ALP ) as well asserumBilirubin (TSB) inT1 group treated with date palm pollen comporting with control groupwhile a significant elevation(P ≤ 0.05) in liver Enzymes (ALT , AST , ALP ) activity and totalserum bilirubin (TSB) in group T2 which exposed to carbon tetrachloride.the protective roleof date palm pollen against carbon tetrachloride was recorded in group T3 which manifestedby significant differences (P&gt;0.05) in liver enzymes activity (ALT , AST , ALP ) and serumbilirubin (TSB) as compared to the control.The histological study of liverof date palm pollentreated rats (group T1) indicated proliferation of kupffer cells and no nuclear lesion, whilegroup T2 showed severe vacuolation in the hepatocytes, while group T3 showed moderatevacuolation in hepatocytes.From the result obtained ,it was concluded that it seems likely thatdosage of rats with (150 mg/kg B.W) of date palm pollen for 42 days of treatment causedimprovement and enhancement of liver function against carbon tetrachloride harmful effects.

STUDY OF HEPATOPROTECTIVE ACTIVITY OF ECBOLIUM VIRIDE (FORSSK.) ALSTON

Objective: To evaluate the hepatoprotective activity of ethanolic extract of Ecbolium viride (Forssk.) Alston i n C C l 4 induced rats. Materials and Methods: The authenticated leaves of Ecbolium viride (E.V) were collected from ABS botanical garden, Salem, Tamil Nadu. The ethanol extract was tested for acute oral toxicity study by using OECD guidelines 423. The starting dose level of Ethanolic extract of E.V was 2000 mg /kg. Body weight p.o. and the route of administration was oral. Dose volume was administered 0.1 ml / 100 gms body weight to the rats, which were fasted over night with water ad libitum. Assessment of hepatoprotective activity was carried out on albino wistar rats. The animals were segregated into five groups of six animals each. Group I served as normal control receiving 5% CMC (10ml/kg). All other groups received CCL4 (1ml/kg) with equal volume of olive oil (50% v/v) for two successive days. Group II animals were maintained as CCL4 group, while group III, IV and V animals were treated orally for seven days with suspension of extract (100mg/kg), extract (200mg/kg) and reference drug silymarin (25 mg/kg), respectively. After the drug treatment all the animals were sacrificed by cervical dislocation. Blood was collected from the carotid artery and was allowed to clot for 45 min at room temperature; serum was separated by centrifugation at 2500 rpm for 15 min, used for the estimation of various biochemical parameters namely GOT, GPT, ALP, ACP and Bilirubin. Biochemical parameters such as Serum Glutamate Pyruvate Transaminase (SGPT), Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Alkaline Phosphatase (SALP), Serum acid Phosphatase (SACP), Liver, and Bilirubin (total & direct) were determined Results: The acute oral toxicity studies showed that ethanolic extract of E.V up to 2000 mg/kg are non toxic and safe. This studies shows that ethanolic extract of E.V possess good hepatoprotective activity. This hepatoprotective activity shown by the E.V may be possibly was due to the active constituent present in these extracts which was responsible for the pharmacological action. Conclusion: The results indicates that ethanolic extract of EV leaves possesses significant h e p a t o p r o t e c t i v e activity.

마늘 추출물이 간 손상 랫드의 조직학적 변화에 미치는 효과 연구

In this study we examined the effects of water extract of garlic on carbon tetrachloride-induced liver injury, and demonstrated increased beneficial enzyme and anti-oxidant activity as well as histopathological changes (by Hematoxylin-Eosin (H&E) staining, Trichrome staining, and TEM examination), and showed that the treatment was dose-dependent and safe. A total of 42 male Sprague-Dawley rats were divided equally (n=7) into six groups. To induce hepatotoxicity in these subjects, carbon tetrachloride diluted in an equal volume of olive oil was intraperitoneally administrated at 0.5 ml/kg (0.20 g/kg/day) once a day for five days. Water extract of Korean-grown garlic was administered via a stomach sonde once a day, 5 days a week, for a total of 4 weeks. Groups received 0.35 g/kg (E1), 0.70 g/kg (E2), or 1.40 g/kg (E3), with the dose adjusted for body weight. Administration of garlic extract resulted in positive physiological effects in terms of reduced oxidative stress and toxicity, and induced functional changes in the liver. Comparing the subject groups (E1, E2, E3) administered different doses of garlic extract, the importance of morphological analysis in further studies is emphasized, because morphological changes indicating hepatotoxicity could occur, even though beneficial enzyme activities were found to be elevated.

Protective effect of retinoid against endotoxin-induced mastitis in rats

A lipopolysaccharide (LPS) induced mastitis model in rats was used to study the protective effect of retinoid. Commencing at gestation day 10, retinoid (dissolved in olive oil) or an equal volume of olive oil was administered to rats daily by gavage until parturition. LPS or pyrogen-free physiological saline was inoculated into the mammary gland 72h after parturition and the rats were euthanized at 12h post-infection. Myeloperoxidase (MPO), N-acetyl -beta-D- glucosaminidase (NAGase), tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8) in mammary tissues and CD4(+)/CD8(+) in peripheral blood were increased and serum MPO and IL-2 in mammary tissues were decreased 12h after LPS infusion. Retinoid decreased MPO, NAGase, and TNF-alpha in mammary tissue and increased IL-2 in serum. Four thousand and 8000 I.U/kg x d of retinoid significantly decreased the infiltration of PMNs in mammary alveoli and ameliorated the imbalance of CD4+/CD8+ in peripheral blood. These results suggest that retinoid protected against LPS-induced mastitis in a rat model.

Retinoid protects rats against neutrophil-induced oxidative stress in acute experimental mastitis

Activated polymorphonuclear neutrophilic leukocytes (PMN) are able to produce large quantities of bactericidal molecules such as reactive oxygen species (ROS) that are associated with tissue damage in models of inflammatory mastitis. In this study, the putative protective effect of retinoid was evaluated in a lipopolysaccharide (LPS) induced mastitis model in rats. Commencing at 10 d of gestation, retinoid (dissolved in olive oil) or an equal volume of olive oil were administered daily by gavage to pregnant rats until parturition. LPS or pyrogen-free physiological saline were infused into the mammary gland 72 h after parturition. At pre-infusion (defined as 0 h) and at 2, 4, 8, 16 and 24 h post-infusion, six rats from each group were euthanized. Retinoid administration decreased PMN accumulation in mammary alveoli, significantly decreased the level of TNF-α in mammary tissues and IL-8 in serum at the different time points. ROS release was significantly increased after LPS infusion and was reduced by retinoid at 16 h PI. Retinoid reduced N-acetyl- β- d-glucosaminidase (NAGase) activity in both serum and mammary tissue at 8 h PI. Intercellular adhesion molecule 1 (ICAM-1) mRNA expression reached its peak value earlier in retinoid treated rats than in the control group. Overall, the results suggest that activated PMN play an important role in the pathogenesis of acute mastitis and retinoid administration may be an effective tool for protecting mammary tissue against PMN-induced oxidative stress during LPS-induced acute mastitis.

REPEATED INTRAPORTAL WHOLE BONE MARROW CELL TRANSPLANTATION IN ANALBUMINEMIC RATS.

P683 Aims: Bone marrow cell (BMC) has been intensively investigated whether these cells could be used for the treatment of liver dysfunctions. However the effectivities of BMC transplantations is still not clear because survival period of transplanted cells were not long enough to support recipient liver functions. We have developed a novel technique which allows repeated infusion of BMC into the portal system via an indwelling catheter. In this study we have developed an effective intraportal transplantation protocol to extend survival time of transplanted cells. Methods: 6weeks old male Nagase Analbuminemic rats (NAR) were injected subcutaneously every 3.5 days with CCl4 dissolved in equal volume of olive oil at a dose of 2.0 ml/kg body weight for 8weeks. Repeated intraportal infusion of 5 x 10(6) rat BMC isolated from 6weeks old female enhanced green fluorescent protein transgenic (EGFP-tg)rat twice a week for the last 4weeks via an indwelling catheter connected to a subcutaneous port. Another NAR underwent repeated intraportal infusion of Dulbecco’s Modified Eagle’s Medium (DMEM). For assessment of liver function, Body and liver weight was measured and blood was obtained for total bilirubin, albumin, PT, transaminase and ammonia. We also evaluated the improvement of fibrosis by histology and repopulating cells by immunohistochemistry. Results: At 4 weeks after the start of BMC transplantation (BMT), we found no significant difference in their total body weight (BMT model: 170g, Control model: 166g). But their liver weight of BMT groups remarkably increased (BMT model: 14.7g, Control model: 11.2g). We also observed macroscopically different between them. Control group liver was white and atrophic, but the liver from BMT model was red and expanded. Microscopic studies revealed that the liver of rat receiving continuous BMT showed decreases in fibrosis, necrosis and inflammatory infiltration. And in the BMT rat, GFP positive cells were found around the portal vein. Prothrombin time(PT), total bilirubin, serum ammonia and transaminase were also significantly better in BMT model compared with control model. Conclusions: Repeated bone marrow cell transplantation via a portal catheter can decrease liver fibrosis and improve liver function of rats with liver cirrhosis and may be useful in the treatment of cirrhosis in humans.

Optimization of the dose and route of injection, and characterisation of the time course of carbon tetrachloride-induced hepatotoxicity in the rat

Introduction: The purpose of this study was to optimize carbon tetrachloride-induced hepatotoxicity in the rat with respect to dose, route of injection, and time course. Methods: Male Wistar albino rats, 4 to 6 weeks old and weighing 130–180 g were used. Hepatotoxicity was evaluated by measuring the activity of serum enzymes (alkaline phosphatase [ALP], alanine aminotransferase [ALT], and aspartate aminotransferase [AST]) as well as serum total bilirubin level. Results: Intraperitoneal injection of carbon tetrachloride (CCl 4) increased the activity of ALP (from 64.9 to 137.3 U/l), ALT (from 106.6 to 693.1 U/l), and AST (from 113.8 to 693.9 U/l). Plasma bilirubin level increased (from 0.119 to 0.42 mg/dl). In contrast, subcutaneous injection of CCl 4 had no effect on these variables. The optimum intraperitoneal dose of CCl 4 was found to be 2 ml/kg body weight (dissolved in an equal volume of olive oil), and this increased the level of bilirubin and the activity of the three enzymes significantly, without causing death of the animals. Hepatotoxicity was observed within 2 h of intraperitoneal injection of CCl 4 and reached a peak after 24 h. Bilirubin level and serum enzyme activities declined gradually to normal levels by 3 days after CCl 4 injection. Conclusion: It is possible to reliably evoke reversible hepatotoxicity in rats by intraperitoneal injection of 2 ml/kg CCl 4.