Statins are HMGCoA reductase inhibitors and had been demonstrated to stimulate bone formation in rodents after high oral doses. Observational studies on patients treated with oral statins were varied. Delta-tocotrienol had been found to stimulate the cleavage of HMGCoA reductase and inhibit its activity. Tocotrienols were found to have both catabolic and anabolic effects on bone in different animal models of osteoporosis. The current study aimed to ascertain the effects of delta-tocotrienol and lovastatin combination on biochemical and static bone histomorphometric parameters in a postmenopausal rat model at clinically tolerable doses. 48 Sprague Dawley female rats were randomly divided into 6 groups: (1) baseline control group; (2) sham-operated control group; (3) ovariectomised control group; (4) ovariectomised and 11 mg/kg lovastatin; (5) ovariectomised and 60 mg/kg delta-tocotrienol; (6) ovariectomised and 60 mg/kg delta-tocotrienol + 11 mg/kg lovastatin. These treatments were given daily via oral gavage for 8 weeks. Delta-tocotrienol plus lovastatin treatment significantly increased bone formation and reduced bone resorption compared to the other groups. Therefore, the combined treatment may have synergistic or additive effects and have the potential to be used as an antiosteoporotic agent in patients who are at risk of both osteoporosis and hypercholesterolemia, especially in postmenopausal women.
Mitragyna speciosa Korth, or better known as ketum, has long been used by traditional folk around Southeast Asia to prevent fatigue from working under hot tropical weather and as a replacement of opium, which can then cause addiction. To date, no findings have been reported of the toxic effect of ketum subchronically (28 days). Hence, the aim of this study was to investigate the toxicity of subchronic effect of standardized methanolic extract of ketum (SMEMS) in Sprague-Dawley rats. Rats were orally administered with 100, 200, and 500 mg/kg of SMEMS for 28 days. Body weights were recorded daily. They were terminated at day 28 to obtain data for hematology, biochemistry, and histopathology of the brain, liver, kidney, lung, heart, sciatic nerve, and spinal cord. The SMEMS affected body weight compared to control group. Biochemistry findings showed that liver and kidney were affected with the abnormal values in AST, creatinine, globulin, glucose, total protein, and urea. However, SMEMS produced toxic effect more to liver, kidney, and lung than other organs as observed histopathologically. The results suggested subchronic exposure of ketum is toxic to the physiology of the animals.
BACKGROUND: Mitragyna speciosa (MS) or ketum is primarily found in Southeast Asia, particularly in northern Malaysia and Thailand. The medicinal value of this plant has attracted significant attention from both herbal medicine practitioners and scientists worldwide. Despite having illegal consumption status, the plant merits study. We conducted a series of experiments to test our hypothesis that ketum impairs both learning and memory in rats.
METHODS: Ketum leaves were extracted using methanol and standardised for the amount of its pure compound, mitragynine. Rats were divided into groups for a passive avoidance task and long-term potentiation (LTP) extracellular recording. In the extracellular recording condition, rats were grouped into control, MS100 (100 mg/kg of ketum extract), MS200 (200 mg/kg of ketum extract), and MS500 (500 mg/kg of ketum extract) groups. An additional group that received morphine was included in the passive avoidance task (10 mg/kg), and there were six animals per group. Rats received daily treatments orally for 28 days for both experiments.
RESULT: Using a passive avoidance task, our data revealed that the rats' memory significantly increased with increasing doses of MS compared to the morphine-treated group. Our findings from LTP recordings showed that LTP was fully blocked by the higher doses of MS.
CONCLUSION: We speculate on the possibility that additional factors were involved in the passive avoidance task because it was an in vivo animal study, while the LTP experiment solely involved brain slices.
Lignosus rhinocerus (L. rhinocerus), which is known locally as Tiger Milk mushroom, is traditionally used in the treatment of asthma by indigenous communities in Malaysia. However, to date, its efficacy on asthma has not been confirmed by scientific studies and there is also sparse information available on its active constituents. In this study, the volatile constituent of L. rhinocerus hot water extract was investigated using gas chromatography mass spectrometry (GC-MS). The potential effects of L. rhinocerus extract for anti-asthmatic activity was further investigated on ovalbumin (OVA)-sensitized asthmatic Sprague Dawley rats.
Aflatoxin B1(AFB1) is a toxic compound commonly found in some crops with an adverse health effect on human and animals. Some beneficial microorganisms (or probiotics) such as lactic acid bacteria have shown the ability to reduce the bioavailability of aflatoxins and its intestinal absorption. However, the dose and duration of aflatoxins exposure and probiotic treatment can influence the ability of probiotics to remove aflatoxins. Therefore, this research aimed to investigate the efficacy of oral probiotic Lactobacillus casei Shirota strain (LcS) induction in an acute exposure to AFB1 in rats. Experimentally, Sprague Dawley rats were divided into three groups: AFB1 only (n = 9); AFB1 treated with LcS (n = 9); and control (no AFB1 exposure) (n = 6) groups. The blood AFB1 level of rats treated with LcS was slightly lower than the untreated AFB1 induced rats (11.12 ± 0.71 vs 10.93 ± 0.69 ng g-1). Also, LcS treatment slightly moderated the liver and kidney biomarkers in AFB1 induced rats. However, a trend for a significant difference was only observed in ALT of AFB1 induced rats treated with LcS compared to their counterparts (126.11 ± 36.90 vs 157.36 ± 15.46, p = 0.06). Rats' body weight decreased in all animals force-fed with AFB1 with no significant difference between LcS treatment compared to the counterpart. In conclusion, this experiment indicated that probiotic LsC was able to slightly ameliorate the adverse effect of an acute exposure to AFB1 in rats. However, future studies with longer probiotics treatment or higher probiotics dose is required to confirm these findings.
Hiperkolesterolemia merupakan suatu keadaan yang dikaitkan dengan perubahan aras lipid serum serta peningkatan aras peroksidaan lipid. Kajian ini dilakukan untuk melihat kesan ekstrak akues isi dan kulit buah Hylocereus polyrhizus (HP) terhadap jumlah kolesterol dan trigliserid (TG) serum serta aras malonaldehid (MDA-TBAR) hati tikus teraruh hiperkolesterolemia. Tikus Sprague dawley jantan diaruh menjadi hiperkolesterolemia dengan pemberian diet rat chow bersama 15% minyak sapi selama 8 minggu dan 0.02 g kolesterol secara suap paksa dua kali seminggu. Tikus-tikus diberi perlakuan ekstrak isi dan kulit buah HP, 300 mg/kg secara suap paksa selama 10 hari. Hasil menunjukkan aras kolesterol menurun secara signifikan (p<0.05) pada kedua-dua kumpulan hiperkolesterolemia yang diberi rawatan ekstrak isi dan kulit buah HP sebanyak 43.53% dan 51.36% berbanding tikus kawalan hiperkolesterolemia. Aras TG menunjukkan penurunan secara signifikan (p<0.05) sebanyak 38.0% bagi kumpulan tikus yang diberi rawatan ekstrak isi dan 42.98% bagi rawatan dengan ekstrak kulit buah HP. Peningkatan aras MDA-TBAR hati telah direncat dengan penurunan aras MDA-TBAR sebanyak 56.85% bagi kumpulan tikus yang diberi ekstrak kulit serta sedikit penurunan iaitu 10.27% bagi tikus yang diberi ekstrak isi berbanding tikus kawalan hiperkolesterolemia. Kajian ini menunjukkan bahawa kedua-dua ekstrak akues isi dan kulit buah HP merendahkan aras lipid serum serta aras MDA-TBAR hati pada tikus teraruh hiperkolesterolemia. Walau bagaimanapun, kesan ekstrak kulit lebih jelas berbanding ekstrak isi yang mungkin disebabkan oleh kandungan betasianin yang lebih tinggi dalam kulit berbanding isi buah HP.
Postmenopausal osteoporosis is one of the main health problems in aging women. It was due to several factors including oxidative stress, which can be controlled through intake of antioxidants from food sources. Virgin coconut oil (VCO) is one of the natural product rich in antioxidants and has been proven to protect osteoporotic bone. This study was conducted to gain in-depth understanding on virgin coconut oil’s activity on osteoporosis at molecular level. Thirty two female Sprague-Dawley rats were divided into four groups, namely Sham operated group, ovariectomized control group (Ovx+Ctrl), ovariectomized with VCO treatment (Ovx+VCO), and ovariectomized with estrogen treatment (Ovx+E). All treatments were administered orally for ten weeks. Bone samples were obtained to examine changes on expression of superoxide dismutase (SOD), glutathione peroxidase (GPX), osteocalcin and runt-related transcription factor 2 (Runx2) genes. The results indicated that rats receiving VCO treatment had experienced significant increments in SOD, GPX and osteocalcin gene expressions compared to the ovariectomized control group, besides the gene expressions of Runx2 which also showed an increment pattern. In conclusion, VCO helps to protect bone in osteoporotic rat model by increasing the expressions of antioxidant genes and genes which increase the osteoblast acitivities.
Keywords: Osteoporosis; ovariectomized rat model; postmenopausal; virgin coconut oil
Cornsilk is traditionally used to treat illnesses related to kidney and as diuretic agent. The study was performed to evaluate the effectiveness of Malaysian cornsilk in elevating diuresis and their dose response relationship in normal Sprague-Dawley rat. The diuresis activity was determined by administered the rats with different dose treatments of 400, 500, 600, 700 and 800 mg/kg. Cumulative urine was significantly increased with the dosage levels (400-600 mg/ kg) ranging from 14.06 - 20.13 mL. Cumulative urine of aqueous extract of cornsilk (AEC) at 400 mg/kg (14.06 mL) and 500 mg/kg (15.21 mL) treatments found to be significantly lower than positive control (21.25 mL). In addition, Na+ content was significantly higher compared with negative control at dosages of 500, 600, 700 and 800 mg/kg. At any rate, K+ and Cl- content of all AEC treatments were not significantly different during 24 h monitoring. The pH values were increased paralleled with the increment of AEC dosages, though it was not significant. On the other result, the ED50 of AEC was observed at 454.10 mg/kg. Malaysian AEC had shown a mild diuretic activity in elevating urine and Na+ content at dosages from 500 to 800 mg/kg. Whilst, AEC also showed an effect of potassium sparing diuretics. Thus, it is suggested that Malaysian cornsilk can be used as an alternative natural diuretic agent.
Tissue recovery is important in preventing tissue deterioration, which is induced by pressure and may lead to pressure ulcers (PU). Reactive hyperaemia (RH) is an indicator used to identify people at risk of PU. In this study, the effect of different recovery times on RH trend is investigated during repetitive loading. Twenty-one male Sprague-Dawley rats (seven per group), with body weight of 385-485 g, were categorised into three groups and subjected to different recovery times with three repetitive loading cycles. The first, second, and third groups were subjected to short (3 min), moderate (10 min), and prolonged (40 min) recovery, respectively, while fixed loading time and pressure (10 min and 50 mmHg, respectively). Peak hyperaemia was measured in the three cycles to determine trends associated with different recovery times. Three RH trends (increasing, decreasing, and inconsistent) were observed. As the recovery time is increased (3 min vs. 10 min vs. 40 min), the number of samples with increasing RH trend decreases (57% vs. 29% vs. 14%) and the number of samples with inconsistent RH trend increases (29% vs. 57% vs. 72%). All groups consists of one sample with decreasing RH trend (14%). Results confirm that different recovery times affect the RH trend during repetitive loading. The RH trend may be used to determine the sufficient recovery time of an individual to avoid PU development.
BACKGROUND: Swietenia macrophylla King. (Meliaceae) seeds (SMS); commonly known as sky fruit and locally known in Malaysia as Tunjuk Langit; have been used in traditional Malay medicine for the treatment of diabetes and hypertension. The people eat only a tiny amount of raw seed, weighing not more than 5 mg.
AIM: To evaluate the safety of Swietenia macrophylla seeds (SMS) at a single-dose oral administration of 2 g/kg body weight (bw) in sprague dawley (SD) rats.
MATERIALS AND METHODS: Eight-week old male and female SD rats were administered a single-oral dose of 2g/kg bw. The rats' general behavior, and toxic signs were observed throughout the 14-day study period. The food and water intake by rats and their body weight were monitored during the study period. At the end of the study period, the relative weights of the organs (lung, liver, spleen, heart, kidney, testis, stomach); the hematological and biochemical parameters were measured; the architecture and histology of the organs (liver, kidney and lungs) were observed.
RESULTS: Oral administration of SMS to rats did not affect, either food or water intake; relative organ weight of vital organs; the hematological and biochemical parameters; did not show significant changes in the architecture and histology of vital organs. Overall, there were neither signs of toxicity nor deaths recorded during the study period.
CONCLUSION: The rat dose of 2 g/kg bw is equivalent to the human dose of 325 mg/kg bw, which is well below the usual amount consumed by people, did not show any signs of toxicity in rats.
KEYWORDS: Diabetes; Swietenia macrophylla; sky fruit; toxicity; traditional Malay medicine; tunjuk langit
Testosterone replacement is the choice of treatment in androgen-deficient osteoporosis. However, long-term use of testosterone is potentially carcinogenic. Eurycoma longifolia (EL) has been reported to enhance testosterone level and prevent bone calcium loss but there is a paucity of research regarding its effect on the bone structural parameters. This study was conducted to explore the bone structural changes following EL treatment in normal and androgen-deficient osteoporosis rat model. Thirty-six male Sprague-Dawley rats aged 12 months were divided into normal control, normal rat supplemented with EL, sham-operated, orchidectomised-control, orchidectomised with testosterone replacement, and orchidectomised with EL supplementation groups. Testosterone serum was measured both before and after the completion of the treatment. After 6 weeks of the treatment, the femora were processed for bone histomorphometry. Testosterone replacement was able to raise the testosterone level and restore the bone volume of orchidectomised rats. EL supplementation failed to emulate both these testosterone actions. The inability of EL to do so may be related to the absence of testes in the androgen deficient osteoporosis model for EL to stimulate testosterone production.
Probiotic Lactobacillus casei Shirota (LcS) is a potential decontaminating agent of aflatoxin B1 (AFB1). However, few studies have investigated the influence of diet, especially a high protein (HP) diet, on the binding of AFB1 by probiotics. This research was conducted to determine the effect of HP diet on the ability of LcS to bind AFB1 and reduce aflatoxin M1 (AFM1) in AFB1-induced rats. Sprague Dawley rats were randomly divided into three groups: A (HP only), B (HP + 108 CFU LcS + 25 μg AFB1/kg BW), and C (HP + 25 μg AFB1/kg BW). Levels of AST and ALP were higher in all groups but other liver function's biomarkers were in the normal range, and the liver's histology showed no structural changes. The urea level of rats in group B (10.02 ± 0.73 mmol/l) was significantly lower (p < 0.05) than that of rats in group A (10.82 ± 0.26 mmol/l). The presence of carcinoma in the small intestine and colon was more obvious in group C than in group B. Moreover, rats in group B had significantly (p < 0.05) lower AFM1 concentration (0.39 ± 0.01 ng/ml) than rats in group C (5.22 ± 0.28 ng/ml). Through these findings, LcS supplementation with HP diet alleviated the adverse effects of AFB1 by preventing AFB1 absorption in the small intestine and reducing urinary AFM1.
Of the twenty microorganisms screened for metabolism of goniothalamin only Streptomyces aurofaciens ATCC 10762 and Nocardia species NRRL 5646 produced two metabolites, 3,4-dihydrogoniothalamin and 3,4,7,8 tetrahydrogoniothalamin. The identity of the isolated metabolites were established using TLC, HPLC, MS, IR, and 1H- and 13C-NMR spectroscopy. In addition, the substrate had been transformed into two unknown metabolites by Aspergillus niger ATCC 11394 and Septomyxa affinis ATCC 6737 in low yield. Three of the metabolites were also detected and identified in the urine and blood samples of the goniothalamin-treated Sprague-Dawley rats. The obtained results are in agreement with and support the principle of microbial models of mammalian metabolism.
The effects of topical application of Orthosiphon stamineus leaf extract on the rate of wound healing and histology of the healed wound were assessed. Four groups of adult male Sprague Dawley rats were experimentally wounded in the posterior neck area. A thin layer of blank placebo was applied topically to wounds of Group 1 rats. Wounds of experimental animals (Group 2 and 3) were dressed with placebo containing 5% and 10% O. stamineus extract, respectively. A thin layer of Intrasite gel® was applied topically to wounds of Group 4 animals as reference. Macroscopically, wounds dressed with placebo containing 5% (healed on day 14.50 ± 0.43) and 10% (healed on day 13.83 ± 0.21) O. stamineus extract each or Intrasite gel® (healed on day 13.13 ± 0.42) significantly accelerated the rate of wound healing compared to wounds dressed with blank placebo. Histological analysis of healed wounds confirmed the results. Wounds dressed with placebo containing 5%, 10% O.stamineus or Intrasite gel® showed markedly less scar width at wound enclosure and granulating tissue contained markedly more collagen, proliferating fibroblast with angiogenesis, and no inflammatory cells compared to wounds dressed with blank placebo. In conclusion, placebo containing 5% or 10% O. stamineus on extract-dressed wounds significantly accelerated the rate of wound healing in rats.
This study investigated the protein quality of two sets of Roselle seeds processed differently (dried and boiled). Twenty weanling Sprague Dawley rats were used to conduct the growth and nitrogen balance studies. Rats were fed with 10% (w/w) protein from dried (DS) and boiled (BS) Roselle seeds powder for 4 weeks. Casein was used in this study as a standard reference protein. There was a significantly higher (p < 0.05) food intake and weight gain by rats fed with BS compared with DS. In the growth study, there was no significant difference (p < 0.05) in protein efficiency ratio (PER) and net protein ratio (NPR) of BS compared to DS, but it was significantly different with casein (CD). PER value of rats fed with DS was significantly lower (p < 0.05) than casein. In the nitrogen balance study, true nitrogen absorption (TNA) and nitrogen balance (NB) of BS group was significantly higher (p < 0.05) than DS group. However, apparent digestibility (AD), true digestibility (TD) and biological value (BV) for both diets was not significantly different. This study showed that the protein quality of dried Roselle seeds was similar to the Roselle seeds boiled at 100oC for 30 minutes.
This study was aimed to evaluate the effect of soursop (Annona muricata L.) extract on Sprague-Dawley rats subjected to in vivo 28-day repeated doses. The extract was given to the study group via force feeding. In the 28-day study, Annona muricata L. extract was dosed at 0 (CD, control dose), 0.5 (LD, low dose), 1.0 (MD, medium dose), 2.0g/kg (HD, high dose) body weight. For control group, distilled water was given to the animals. Administration of Annona muricata L. extract did not cause negative effect in blood hematology even though a statistically significant (p
Spinal cord injury (SCI) is a devastating disorder that has a poor prognosis of recovery. Animal models of SCI are useful to understand the pathophysiology of SCI and the potential use of therapeutic strategies for human SCI. Ex vivo models of central nervous system (CNS) trauma, particularly mechanical trauma, have become important tools to complement in vivo models of injury in order to reproduce the sequelae of human CNS injury. Ex vivo organotypic slice cultures (OSCs) provide a reliable model platform for the study of cell dynamics and therapeutic intervention following SCI. In addition, these ex vivo models support the 3R concept of animal use in SCI research - replacement, reduction and refinement. Ex vivo models cannot be used to monitor functional recovery, nor do they have the intact blood supply of the in vivo model systems. However, the ex vivo models appear to reproduce many of the post traumatic events including acute and secondary injury mechanisms. Several well-established OSC models have been developed over the past few years for experimental spinal injuries ex vivo in order to understand the biological response to injury. In this study, we investigated cell viability in three ex vivo OSC models of SCI: stab injury, transection injury and contusion injury. Injury was inflicted in postnatal day 4 rat spinal cord slices. Stab injury was performed using a needle on transverse slices of spinal cord. Transection injury was performed on longitudinal slices of spinal cord using a double blade technique. Contusion injury was performed on longitudinal slices of spinal cord using an Infinite Horizon impactor device. At days 3 and 10 post-injury, viability was measured using dual staining for propidium iodide and fluorescein diacetate. In all ex vivo SCI models, the slices showed more live cells than dead cells over 10 days in culture, with higher cell viability in control slices compared with injured slices. Although no change in cell viability was observed between time-points in stab- and contusion-injured OSCs, a reduction in cell viability was observed over time in transection-injured OSCs. Taken together, ex vivo SCI models are a useful and reliable research tool that reduces the cost and time involved in carrying out animal studies. The use of OSC models provides a simple way to study the cellular consequences following SCI, and they can also be used to investigate potential therapeutics regimes for the treatment of SCI.
The effects of Eurycoma longifolia Jack were studied on the libido of sexually experienced male rats after dosing them with 200, 400 and 800 mg/kg body weight twice daily of different fractions of E. longifolia Jack for 10 days. Results showed that E. longifolia Jack produced a dose-dependent increase in mounting frequency of the treated animals with 400 mg/kg of chloroform, methanol, water and butanol fractions resulting in mounting frequencies of 5.3 +/- 1.2, 4.9 +/- 0.7, 4.8 +/- 0.7 and 5.2 +/- 0.1, and 800 mg/kg further increased them to 5.4 +/- 0.8, 5.4 +/- 0.8, 5.2 +/- 0.6 and 5.3 +/- 0.2 respectively but there were no erections, intromissions, ejaculations or seminal emissions during the 20-min observation period which allowed for the measurement of sexual arousal reflected by mounting frequency uninfluenced by other behavioural components. This study provides evidence that E. longifolia Jack is a potent stimulator of sexual arousal in sexually vigorous male rats in the absence of feedback from genital sensation.
Recent evidence has exhibited dietary influence on the manifestation of different types of behavior induced by stressor tasks. The present study examined the effects of Tualang honey supplement administered with the goal of preventing or attenuating the occurrence of stress-related behaviors in male rats subjected to noise stress. Forty-eight adult male rats were randomly divided into the following four groups: i) nonstressed with vehicle, ii) nonstressed with Tualang honey, iii) stressed with vehicle, and iv) stressed with honey. The supplement was given once daily via oral gavage at 0.2 g/kg body weight. Two types of behavioral tests were performed, namely, the novel object recognition test to evaluate working memory and the forced swimming test to evaluate depressive-like behavior. Data were analyzed by a two-way analysis of variance (ANOVA) using IBM SPSS 18.0. It was observed that the rats subjected to noise stress expressed higher levels of depressive-like behavior and lower memory functions compared to the unexposed control rats. In addition, our results indicated that the supplementation regimen successfully counteracted the effects of noise stress. The forced swimming test indicated that climbing and swimming times were significantly increased and immobility times significantly decreased in honey-supplemented rats, thereby demonstrating an antidepressant-like effect. Furthermore, cognitive function was shown to be intensely affected by noise stress, but the effects were counteracted by the honey supplement. These findings suggest that subchronic exposure to noise stress induces depressive-like behavior and reduces cognitive functions, and that these effects can be attenuated by Tualang honey supplementation. This warrants further studies to examine the role of Tulang honey in mediating such effects.
Aging is characterized by progressive decline in physiological and body function due to increase in oxidative damage. Gelam honey has been accounted to have high phenolic and nonphenolic content to attenuate oxidative damage. This study was to determine the effect of local gelam honey on oxidative damage of aged rats. Twenty-four male Spraque-Dawley rats were divided into young (2 months) and aged (19 months) groups. Each group was further divided into control (fed with plain water) and supplemented with 2.5 mg/kg body weight of gelam honey for 8 months. DNA damage level was determined by comet assay and plasma malondialdehyde (MDA) by high performance liquid chromatography (HPLC). The activity of blood and cardiac antioxidant enzymes was determined by spectrophotometer. The DNA damage and MDA level were reduced in both gelam honey supplemented groups. Gelam honey increases erythrocytes CAT and cardiac SOD activities in young and cardiac CAT activity in young and aged groups. The DNA damage was increased in the aged group compared to young group, but reduced at the end of the study. The decline of oxidative damage in rats supplemented with gelam honey might be through the modulation of antioxidant enzyme activities.