Studies on the Antiulcerogenic and Antihepatotoxic Potential of Chitosan and Glucosamine in Experimental Rats

The present study examined the antiulcer effect of glucosamine on mucosal antioxidant defense system in ibuprofen-induced peptic ulcer in male albino rats. The results of the present study indicate that the pre-oral administration of chitosan and glucosamine maintain near to the normal status the activities of the mucosal antioxidant enzymes and the level of GSH (Glutathione), which protect mucosa against oxidative damage by decreasing the lipid peroxidation and strengthening the mucosal barrier, and which are the first line of defense against exogenous ulcerogenic agents. In this study indicate that the oral pre-treatment of chitosan and glucosamine can prevent ibuprofen-induced peptic ulcer in rats.This study can be concluded that co-administration of chitsosan and glucosamine can effectively prevent the isonized and rifampicin induced hepatotoxicity in rats.Comparatively, chitosan was found to have better results than glucosamine in alleviating the hepatic disorders.

Biochemical Effect of Dicarboxylic Acids on Oxalate Metabolism in Experimental Rats and Studies on Oxalate Degrading Bacteria

The current study is an attempt to find a means of lowering oxalate concentration in individuals susceptible to recurrent calcium oxalate stone disease.The formation of renal stone composed of calcium oxalate is a complex process that remains poorly understood and treatment of idiopathic recurrent stone formers is quite difficult and this area has attracted lots of research workers. The main objective of this work are to study the effect of certain mono and dicarboxylic acids on calcium oxalate crystal growth in vitro, isolation and characterization of oxalate degrading bacteria, study the biochemical effect of sodium glycollate and dicarboxylic acids on oxalate metabolism in experimental stone forming rats and To investigate the effect of dicarboxylic acids on oxalate metabolism in experimental hyperoxaluric rats. Oxalic acid is one of the most highly oxidized organic compound widely distributed in the diets of man and animals, and ingestion of plants that contain high concentration of oxalate may lead to intoxication. Excessive ingestion of dietary oxalate may lead to hyperoxaluria and calcium oxalate stone disease.The formation of calcium oxalate stone in the urine is dependent on the saturation level of both calcium and oxalate. Thus the management of one or both of these ions in individuals susceptible to urolithiasis appears to be important. The control of endogenous oxalate synthesis from its precursors in hyperoxaluric situation is likely to yield beneficial results and can be a useful approach in the medical management of urinary stones. A variety of compounds have been investigated to curtain endogenous oxalate synthesis which is a crucial factor, most of these compounds have not proved to be effective in the in vivo situation and some of them are not free from the toxic effect. The non-operative management of stone disease has been practiced in ancient India in the three famous indigenous systems of medicine, Ayurveda, Unani and Siddha, and proved to be effective.However the efficiency of most of these substances is still questionable and demands further study. Man as well as other mammals cannot metabolize oxalic acid. Excessive ingestion of oxalic acid can arise from oxalate rich food and from its major metabolic precursors, glycollate, glyoxylate and ascorbic acid can lead to an acute oxalate toxicity. Increasedlevels of circulating oxalate, which can result in a variety of diseases including renal failure and oxalate lithiasis. The ability to enzymatically degrade oxalate to less noxious Isubstances, formate and CO2, could benefit a great number of individuals including those afflicted with hyperoxaluria and calcium oxalate stone disease.

Antilipolytic and hypotriglyceridemic effects of dietary Salvia triloba Lf (Lamiaceae) in experimental rats

Purpose: Pancreatic triacylglycerol lipase (PL) is a noteworthy pharmacological target for the management of dyslipidemia, and diabetes and obesity. This study was aimed to evaluate the modulatory effects of Salvia triloba L.f. (Lamiaceae) leaves methanol extract (ME) on a high fat diet (HFD)-induced hypertriglyceridemia in rats, with complementary in vitro evaluation of sage PL-inhibitory potential. Methods: Pre-induction of HFD hypertriglyceridemia sage leaves ME (750 mg/kg) was orally supplemented (via gastric intubation) to overnight fasting rats (n = 5). Potential plant modulation of PL was also quantified in vitro by a colorimetric assay (n = 3). For comparison, the effect of Orlistat was similarly evaluated as reference standard. Results: Compared to Orlistat, supplementation of S. triloba at a dose of 750 mg/kg b.wt significantly reversed the HFD-induced postprandial hypertriglyceridemia in experimental overnight fasting rats (p < 0.001 vs. HFD rats). Dietary sage caused 66.4 % reduction in plasma triglycerides. Compared to Orlistat which exerted antilipolytic activity, with half-maximal inhibitory concentration (IC50) of 0.114 ± 0.004 μg/mL), sage inhibited PL activity in vitro in a dose-dependent manner IC50 of 100.80 ± 9.07 μg/mL) Conclusion: Sage has dual hypotriglyceridemic and antilipolytic properties which indicate that it can potentially be used to suppress body weight gain.

Anti-osteoporosis activity of Astragalus membranaceus Bunge extract in experimental rats

Purpose: To investigate the anti-osteoporosis effect of Astragalus membranaceus (Fisch.) Bunge. extract (AMBE) in experimental rats. Method: Female Sprague-Dawley rats were randomly divided into six groups: control group, ovariectomy (OVX) with vehicle group, OVX with 17β-estradiol (E2, 25 μg/kg/day) group, and OVX with AMBE doses (60, 120 and 240 mg/kg/day) groups. Daily oral administration of AMBE or E2 was started 4 weeks after OVX and lasted for 16 weeks. The bone mineral density (BMD) of L4 vertebrae and right femurs was evaluated. The length of each femur was measured with a micrometer, and the center of diaphysis was determined. Three representative L4 vertebrae were selected to evaluate trabecular microarchitecture. Serum alkaline phosphatase (ALP), urinary calcium (U-Ca), urinary phosphorus (UP), urinary creatinine (Cr) and osteocalcin (OC) levels were measured. Results: AMBE dose-dependently inhibited the bone mineral density (BMD) reduction of L4 vertebrae (0.27 ± 0.03 g/cm2, p < 0.05) and femurs (0.23 ± 0.03 g/cm2, p < 0.05) caused by OVX and prevented the deterioration of trabecular microarchitecture (p < 0.05), which were accompanied by a significant decrease in skeletal remodeling (p < 0.05) as evidenced by the lower levels of bone turnover markers. A higher dosage of AMBE treatment (240 mg/kg/day) increased U-Ca/Cr (0.27 ± 0.03 mmol/mmol), ALP (137.23 ± 16.72 U/L), U-P/Cr (4.18 ± 0.27 mmol/mmol) and OC (8.47 ± 0.26 mmol/L) levels (both p < 0.05). Conclusion: The findings of this study indicate that AMBE prevents OVX-induced osteoporosis in rats.

Ultrastructural study of the lateral lobe of the prostate of Wistar rats submitted to experimental chronic alcohol ingestion

Alcoholism is considered today one of the most serious social and medical problems. Different investigators have demonstrated that chronic exposure to alcohol causes morphological and physiological changes in the testes and in the accessory sex glands, in the prostate in particular. In the present study, experimental rats were divided into groups receiving sugar cane brandy diluted to 30° G.L. (30%, v/v) and Purina ration ad libitum for 60, 120, 180, 240 and 300 days, respectively. At the end of each period the animals were sacrificed and the lateral lobe of the prostate was collected for histological examination. The results showed atrophied epithelium, accumulation of lipidic droplets and rupture of the microvilli that line the cell surface facing the lumen of cells of the secretory epithelium of the lateral lobe of the prostate.

Evaluation of anti-obesity activities of ethanolic extract of Terminalia paniculata bark on high fat diet-induced obese rats

Background: The prevalence and severity of obesity and associated co-morbidities are rapidly increasing across the world. Natural products-based drug intervention has been proposed as one of the crucial strategies for management of obesity ailments. This study was designed to investigate the anti-obesity activities of ethanolic extract of Terminalia paniculata bark (TPEE) on high fat diet-induced obese rats. Methods: LC-MS/MS analysis was done for ethanolic extract of T. paniculata bark. Male Sprague-Dawley (SD) rats were randomly divided into six groups of six each, normal diet fed (NC), high fat diet-fed (HFD), HFD+ orlistat (standard drug control) administered, and remaining three groups were fed with HFD + TPEE in different doses (100,150 and 200 mg/kg b. wt). For induction of obesity rats were initially fed with HFD for 9 weeks, then, (TPEE) was supplemented along with HFD for 42 days. Changes in body weight, body composition, blood glucose, insulin, tissue and serum lipid profiles, atherogenic index, liver markers, and expression of adipogenesis-related genes such as leptin, adiponectin, FAS, PPARgamma, AMPK-1alpha and SREBP-1c, were studied in experimental rats. Also, histopathological examination of adipose tissue was carried out. Results: Supplementation of TPEE reduced significantly (P < 0.05) body weight, total fat, fat percentage, atherogenic index, blood glucose, insulin, lipid profiles and liver markers in HFD-fed groups, in a dose-dependent manner. The expression of adipogenesis-related genes such as Leptin, FAS, PPARgamma, and SREBP-1c were down regulated while Adiponectin and AMPK-1alpha were up regulated in TPEE + HFD-fed rats. Furthermore, histopathological examination of adipose tissue revealed the alleviating effect of TPEE which is evident by reduced size of adipocytes. Conclusions: Together, the biochemical, histological and molecular studies unambiguously demonstrate the potential anti adipogenic and anti obesity activities of TPEE promoting it as a formidable candidate to develop anti obesity drug.

Decreased a2-adrenergic receptor in the brain stem and pancreatic islets during pancreatic regeneration in weanling rats

Sympathetic stimulation inhibits insulin secretion. a2-Adrenergic receptor is known to have a regulatory role in the sympathetic function. We investigated the changes in the a2-adrenergic receptors in the brain stein and pancreatic islets using [3H]Yohimbine during pancreatic regeneration in weanling rats. Brain stem and pancreatic islets of experimental rats showed a significant decrease (p<0.001) in norepinephrine (NE) content at 72 h after partial pancreatectomy. The epinephrine (EPI) content showed a significant decrease (p<0.001) in pancreatic islets while it was not detected in brain stem at 72 h after partial pancreatectomy. Scatchard analysis of [3H]Yohimbine showed a significant decrease (p<0.05) and Kd at 72 h after partial pancreatectomy in the brain stem. In the pancreatic islets, Scatchard analysis of [3H]Yohimbine showed a signiinfiBca'nnatx decrease (p<0.001) in B,nax and Kd (p<0.05) at 72 h after partial pancreatectomy. The binding parameters reversed to near sham by 7 days after pancreatectomy both in brain stein and pancreatic islets. This shows that pancreatic insulin secretion is influenced by central nervous system inputs from the brain stem. In vitro studies with yohimbine showed that the a2-adrenergic receptors are inhibitory to islet DNA synthesis and insulin secretion. Thus our results suggest that decreased a2-adrenergic receptors during pancreatic regeneration functionally regulate insulin secretion and pancreatic 13-cell proliferation in weanling rats.

GABA, serotonin, muscarinic receptors - their second messengers and transcription factors functional regulation in the brain regions of hypoxic neonatal rats: Resuscitation with glucose, oxygen and epinephrine

The present study was designed to investigate the protective effect of glucose, oxygen and epinephrine resuscitation on impairment in the functional role of GABAergic, serotonergic, muscarinic receptors, PLC, BAX, SOD, CAT and GPx expression in the brain regions of hypoxia induced neonatal rats. Also, the role of hormones - Triiodothyronine (T3) and insulin, second messengers – cAMP, cGMP and IP3 and transcription factors – HIF and CREB in the regulation of neonatal hypoxia and its resuscitation methods were studied. Behavioural studies were conducted to evaluate the motor function and cognitive deficit in one month old control and experimental rats. The efficient and timely supplementation of glucose plays a crucial role in correcting the molecular changes due to hypoxia, oxygen and epinephrine. The sequence of glucose, epinephrine and oxygen administration at the molecular level is an important aspect of the study. The additive neuronal damage effect due to oxygen and epinephrine treatment is another important observation. The corrective measures by initial supply of glucose to hypoxic neonatal rats showed from the molecular study when brought to practice will lead to healthy intellectual capacity during the later developmental stages, which has immense clinical significance in neonatal care.

Glutamatergic Nmda And Ampa Receptors Functional Regulation In Streptozotocin Induced Diabetic Rats: Effect Of Vitamin D3 And Curcumin Supplementation

The present study was designed to investigate the protective effect of curcumin and vitamin D3 in the functional regulation of glutamatergic NMDA and AMPA receptors in streptozotocin (STZ) induced diabetic rats. Alterations in glutamatergic neurotransmission in the brain were evaluated by analyzing the glutamate content, glutamate receptors - NMDA and AMPA receptors binding parameters and gene expression, GAD and GLAST gene expression. Immunohistochemistry studies using confocal microscope were carried out to confirm receptor density and gene expression results of NMDA and AMPA receptors. The role of glutamatergic receptors in pancreas was studied using the following parameters; glutamate content, GLAST expression, glutamate receptors - NMDA and AMPA receptor binding and gene expression. Increasing evidence in both experimental and clinical studies suggests that oxidative stress plays a major role in the pathogenesis of diabetes. In the present study SOD assay and GPx gene expression were done to evaluate the activity of antioxidant enzymes in the brain regions and pancreas. NeuroD1 and Pdx1 gene expression were performed in pancreas of experimental rats to evaluate pancreatic islet survival. Gene expression profiles of caspase 8, Bax, and Akt in brain regions and pancreas were studied to understand the possible mechanism behind curcumin and vitamin D3 mediated neuroprotection and islet survival. Gene expression studies of vitamin D3 receptor localisation in the pancreas was done to understand the mechanism of vitamin D3 in insulin secretion. Curcumin and vitamin D3 mediated insulin secretion via Ca2+ release were studied using confocal microscope.

Repair of the anophthalmic cavity of rats with synthetic hydroxyapatite

We placed spheres of synthetic hydroxyapatite (calcium chloride combined with sodium phosphate) in the eviscerated or enucleated orbital cavity of rats in order to evaluate the biocompatibility of this material with the orbital cavity. The study was conducted on 50 albino rats, 25 of which were submitted to enucleation and 25 to evisceration of one eye. The animals were sacrificed 7, 15, 21, 30 and 60 days after surgery and the orbital content was submitted to histopathological examination. A reaction of the young granulation tissue type was observed first. The hydroxyapatite was gradually surrounded by a granulomatous macrophage inflammatory response and covered with dense connective tissue that formed a sort of mesh septating and supporting progressively smaller blocks of the substance. The same type of reaction was observed in the enucleated and eviscerated cavities. We conclude that synthetic hydroxyapatite is an inert nonallergenic material which is appropriate for volume replacement in the anophthalmic cavity

Hepatoprotective effect of water soluble extract of Coleus barbatus on cholestasis on young rats

OBJETIVO: Testar os efeitos do extrato aquoso de Coleus barbatus (EAB) na cirrose biliar secundária por obstrução das vias biliares extra-hepáticas em ratos jovens. MÉTODOS: Quarenta ratos Wistar machos com 21 dias de vida (P21), foram distribuídos em quatro grupos de 10 animais, submetidos a operação simulada ou dupla ligadura e ressecção do ducto biliar (S ou L) combinados EAB e a Água (B ou A). No P48, foi medido o tempo de sono com o pentobarbital (TS). No P49, foram submetidos a eutanásia para a determinação das atividades séricas do aspartato aminotransferase (AST) e da alanina aminotransferases (ALT); após a eutanásia foram avaliados o peso fresco do fígado (PFF) e, em cortes histológicos do fígado, a freqüência de mitoses (FM), o número de áreas de necrose (NN), a intensidade da fibrose (IF) e da proliferação ductal (IPD). Os efeitos da colestase, os do EAB e suas interações foram testados pela ANOVA com dois fatores, e as comparações múltiplas pareadas foram realizadas pelo teste de S.N.K ou teste de Wilcoxon. Também foi determinada a correlação linear de Pearson entre as variáveis histológicas duas a duas separadamente para os grupos LA e LD. O nível de significância estatística para os vários testes foi de p do erro alfa <0,05. RESULTADOS: A colestase aumentou significativamente o TS, a ALT, a AST, o PFF, a MI, o NN, a IF e a IPD. O EAB diminuiu o TS e a IM nos animais sem colestase (operação simulada). O EAB diminuiu o TS, a ALT, a AST, o PFF, a MI, o NN e IF na colestase. No grupo LA houve correlação positiva entre a IPD e a IF, correlação negativa entre a IPD e a FM e correlação negativa entre a IF a FM. No grupo LD houve correlação negativa entre o NN e a IPD. CONCLUSÕES: Na ausência de colestase o EAB encurta o tempo de sono e diminui a freqüência de mitoses. O EAB apresenta efeito hepatoprotetor no modelo de cirrose biliar secundária a obstrução biliar extra-hepática.

O RATO COMO MODELO EXPERIMENTAL DO DIABETES - ESTUDO CLINICO-LABORATORIAL

In this study, 90 Wistar rats were used. They were equally divided into three experimental groups - control group (CG), diabetic group (DG) and treated diabetic group (TG). The analyzed parameters were clinical (behavior, activity, general aspect, weight, water ingestion and diuresis) and biochemical (fasting glycemia and urinary glycosis). The diabetes was induced by alloxan and, then, treated with insulin associated to oral hypoglycemic (acarbosis). Observations were made at 5 experimental moments, as it follows: 1, 3, 6, 9, and 12 months after the diabetes induction. The results were submitted to variance analysis, with 5% of significance level. The DG presented lower weight and higher diuresis level than the CG and TG. The water ingestion of the CG was similar to TG. The glycemia levels were higher in DG than in CG, at every experimental moment. The TG, however, presented glycemia similar to the CG, except for the dosages at 3, and 9 months. They urinary glycosis of the DG and TG were similar between themselves, but higher than the one of the CG.

Estudo da carcinogênese bucal experimental utilizando-se o óxido de nitroquinolina (4-NQO) em ratos

Pós-graduação em Odontologia - FOAR

Blockade of estrogen synthesis with an aromatase inhibitor affects luteal function of the pseudopregnant rat

The luteotropic action of estrogen (E) was investigated using immature pseudopregnant rat as the model and CGS 16949A (Fadrozole hydrochloride), a potent aromatase inhibitor (AI), to block E synthesis. Aromatase activity could be inhibited by administering CGS 16949A (50 mu g/day/rat) via a mini osmotic Alzet pump (model 2002) for 3 days during pseudopregnancy. This resulted in significant reduction of serum (40%, P < 0.05) and intraovarian (70.6%, P < 0.001) estradiol-17 beta (E(2)) levels. The serum and intraovarian progesterone (P-4) levels as analyzed on day 4 of pseudopregnancy were also reduced by greater than or equal to 50% (for both, P < 0.01). Simultaneous administration of estradiol-3-benzoate (E(2)B) via an Alzet pump during the Al: treatment period at a dose of 1 mu g/day could completely reverse the Al induced reduction in P-4 secretion. The luteal cells of experimental rats depleted of E in vivo showed a significantly reduced response upon incubation with hCG or dbcAMP in vitro (P < 0.05 and 0.001, respectively). Addition of E(2) (500 pg/tube) at the time of in vitro incubation was able to partially increase the responsiveness to hCG. The luteal cell LH/hCG receptor content and the affinity of hCG binding to the receptor remained unchanged following AI treatment in vivo. Both esterified and total cholesterol content of luteal cells of rats treated with Al in vivo was significantly high (P < 0.05) suggesting that E lack results in an impairment in cholesterol utilization for steroidogenesis. The results clearly show that E regulates luteal function in the pseudopregnant rat by acting at a non-cAMP mediated event and this perhaps involves facilitation of cholesterol utilization at the mitochondrial level for P-4 synthesis.

Adrenergic and muscarinic receptor subtypes functional regulation during diabetogenesis: Effect of curcumin and vitamin D3 pre-treatment

In the present study, the initial phase was directed to confirm the effects of curcumin and vitamin D3 in preventing or delaying diabetes onset by studying the blood glucose and insulin levels in the pre-treated and diabetic groups. Behavioural studies were conducted to evaluate the cognitive and motor function in experimental rats. The major focus of the study was to understand the cellular and neuronal mechanisms that ensure the prophylactic capability of curcumin and vitamin D3. To elucidate the mechanisms involved in conferring the antidiabetogenesis effect, we examined the DNA and protein profiles using radioactive incorporation studies for DNA synthesis, DNA methylation and protein synthesis. Furthermore the gene expression studies of Akt-1, Pax, Pdx-1, Neuro D1, insulin like growth factor-1 and NF-κB were done to monitor pancreatic beta cell proliferation and differentiation. The antioxidant and antiapoptotic actions of curcumin and vitamin D3 were examined by studying the expression of antioxidant enzymes - SOD and GPx, and apoptotic mediators like Bax, caspase 3, caspase 8 and TNF-α. In order to understand the signalling pathways involved in curcumin and vitamin D3 action, the second messengers, cAMP, cGMP and IP3 were studied along with the expression of vitamin D receptor in the pancreas. The neuronal regulation of pancreatic beta cell maintenance, proliferation and insulin release was studied by assessing the adrenergic and muscarinic receptor functional regulation in the pancreas, brain stem, hippocampus and hypothalamus. The receptor number and binding affinity of total muscarinic, muscarinic M1, muscarinic M3, total adrenergic, α adrenergic and β adrenergic receptor subtypes were studied in pancreas, brain stem and hippocampus of experimental rats. The mRNA expression of muscarinic and adrenergic receptor subtypes were determined using Real Time PCR. Immunohistochemistry studies using confocal microscope were carried out to confirm receptor density and gene expression results. Cell signalling alterations in the pancreas and brain regions associated with diabetogenesis and antidiabetogenesis were assessed by examining the gene expression profiles of vitamin D receptor, CREB, phospholipase C, insulin receptor and GLUT. This study will establish the anti-diabetogenesis activity of curcumin and vitamin D3 pre-treatment and will attempt to understand the cellular, molecular and neuronal control mechanism in the onset of diabetes.Administration of MLD-STZ to curcumin and vitamin D3 pre-treated rats induced only an incidental prediabetic condition. Curcumin and vitamin D3 pretreated groups injected with MLD-STZ exhibited improved circulating insulin levels and behavioural responses when compared to MLD-STZ induced diabetic group. Activation of beta cell compensatory response induces an increase in pancreatic insulin output and beta cell mass expansion in the pre-treated group. Cell signalling proteins that regulate pancreatic beta cell survival, insulin release, proliferation and differentiation showed a significant increase in curcumin and vitamin D3 pre-treated rats. Marked decline in α2 adrenergic receptor function in pancreas helps to relent sympathetic inhibition of insulin release. Neuronal stimulation of hyperglycemia induced beta cell compensatory response is mediated by escalated signalling through β adrenergic, muscarinic M1 and M3 receptors. Pre-treatment mediated functional regulation of adrenergic and cholinergic receptors, key cell signalling proteins and second messengers improves pancreatic glucose sensing, insulin gene expression, insulin secretion, cell survival and beta cell mass expansion in pancreas. Curcumin and vitamin D3 pre-treatment induced modulation of adrenergic and cholinergic signalling in brain stem, hippocampus and hypothalamus promotes insulin secretion, beta cell compensatory response, insulin sensitivity and energy balance to resist diabetogenesis. Pre-treatment improved second messenger levels and the gene expression of intracellular signalling molecules in brain stem, hippocampus and hypothalamus, to retain a functional neuronal response to hyperglycemia. Curcumin and vitamin D3 protect pancreas and brain regions from oxidative stress by their indigenous antioxidant properties and by their ability to stimulate cellular free radical defence system. The present study demonstrates the role of adrenergic and muscarinic receptor subtypes functional regulation in curcumin and vitamin D3 mediated anti-diabetogenesis. This will have immense clinical significance in developing effective strategies to delay or prevent the onset of diabetes.