Rats In Control Group Research Articles

Effect of Long-Term Adaptation to Cold and Short-Term Cooling on the Expression of the TRPM2 Ion Channel Gene in the Hypothalamus of Rats

The present study is aimed to elucidate the possible involvement of the thermosensitive TRPM2 ion channel in changing of the temperature sensitivity of the hypothalamus after different cold exposures-long-term adaptation to cold and short-term cooling. Quantitative RT-PCR was used to study the expression of the gene of thermosensitive TRPM2 ion channel in the hypothalamus in the groups of control (kept for 5 weeks at +20 to +22 °C) and cold-adapted (5 weeks at +4 to +6 °C) rats, as well as in the groups of animals which were subjected to acute cooling (rapid or slow) with subsequent restoration of body temperature to the initial level. It has been shown that after long-term adaptation to cold, the decrease in the Trpm2 gene expression was observed in the hypothalamus, while a short-term cooling does not affect the expression of the gene of this ion channel. Thus, long-term adaptation to cold results in the decrease in the activity not only of the TRPV3 ion channel gene, as shown earlier, but also of the Trpm2 gene in the hypothalamus. The overlapping temperature ranges of the functioning of these ion channels and their unidirectional changes during the adaptation of the homoeothermic organism to cold suggest their functional interaction. The decrease in the Trpm2 gene expression may indicate the participation of this ion channel in adaptive changes in hypothalamic thermosensitivity, but only as a result of long-term cold exposure and not of a short-term cooling. These processes occurring at the genomic level are one of the molecular mechanisms of the adaptive changes.

Effects of Toll-Like Receptor4 Gene Modified Bone Marrow Mesenchymal Stem Cells (BMSCs) Transplantation on Expression of Basic Fibroblast Growth Factor (bFGF) and Follistatin-Like-1 (FSTL1) in Myocardial Ischemia-Reperfusion Rats

This study assesses the effects of TLR4 gene modified BMSCs transplantation on the expression of bFGF and FSTL1 in myocardial ischemia-reperfusion rats. 30 male SD rats were assigned into control group (myocardial ischemia model), BMSCs group (model + BMSCs transplantation) and transfection group (model + TLR4 gene modified BMSCs transplantation) followed by analysis of TLR4 expression, EGFP, apoptosis and expression of bFGF and FSTL1. Compared with control group (TLR4 concentration 2.86 pg/5×105 cells/mL). The expression of TLR4 in BMSCs group (25.24 pg/5×105 cells/mL) and transfection group (31.55 pg/5×105 cells/mL) was significantly increased (P <0.05), and it was more significant in transfection group. The myocardial tissue of rats in control group produced a large number of scars, hypertrophy and hyperplasia of myocardial cells accompanied by a large number of necrosis; The scar tissue of the myocardium in BMSCs group and transfection group decreased, and viable myocardium increased, with more significant effect in transfection group. Control group showed a large number of blue collagen fibers in the infarction area of left ventricle, which were in the shape of cords, and part of the collagen fibers were fused. The blue collagen fibers in the control group and the transfection group were significantly reduced. Compared with control group, BMSCs group had lower apoptosis, and increased bFGF and FSTL1 levels (P <0.05). Compared with BMSCs group, the apoptosis rate of myocardial cells was decreased, and the levels of bFGF and FSTL1 were increased (P < 0.05). In conclusion, transplantation of BMSCs modified with TLR4 can increase bFGF and FSTL1 levels, reduce the rate of myocardial apoptosis and improve the myocardial pathological tissue, thus playing a therapeutic role.

Immunohistochemical determination of somatostatin release in gastric tissue of rats fed with a high-fat and cholesterol diet.

This study aimed to investigate the effects of a high-fat and cholesterol diet (HFCD) on rats' gastric mucosa. In the study, a total of 16 (40-day-old Sprague Dawley) male rats were used and randomly divided into two groups (each consisted of eight rats). The rats in control group had no implementations other than normal feeding. For 10 weeks, rats in a high-fat with cholesterol diet group had daily energy amounts provided by pellet feed mixed with 65.00% butter and 2.00% cholesterol. Before beginning the study and at the end, rats live weight was recorded and their blood samples were taken for biochemical analyses. Hematoxylin and Eosin and Crossman's triple staining techniques were used to investigate the general structure of gastric tissue. The rats fed with HFCD had statistically significant increases in live weight and total cholesterol values, and were identified to have gastric tissue degeneration. The rats' gastric tissue in control group had more intense somatostatin (SST) immunoreactivity in parietal and chief cells than the HFCD group. It was determined that feeding with the HFCD has a negative effect on SST secretion in rats and hence, this may have important areas of use such as in gastric cancer treatment and preventing complications linked to gastric diseases.

Effects of lipopolysaccharide stimulation during pregnancy on kidneys and blood vessels in offspring rats

Objective: To investigate the effect of lipopolysaccharide (LPS) inflammatory stimulation during pregnancy on offspring rats. Methods 20 SD pregnant rats were randomly divided into control and LPS groups, with 10 rats in each group. Rats in LPS group received intraperitoneal injection of LPS 0.79 mg/kg on the Days 8th, 10th, and 12th of gestation, and rats in control group received intraperitoneal injection of the same volume of sterile saline. The blood pressure of offspring rats was measured once every two weeks from 8 weeks of age to 16 weeks of age. Protein expression of NF-κB and NOX2 was detected by immunohistochemistry. ROS and NO levels in vascular tissues were observed by staining with DHE and DAF-2DA cular tissues were observed by staining with DHE and DAF-2DA fluorescent probes, respectively. Results At 8, 10, 12, 14 and 16 weeks of age,the systolic blood pressure of offspring in the LPS group was higher than that of the control group at the same age (P< 0.01).The urine volume and urinary sodium excretion rate of the LPS group were significantly lower than those of the control group (P < 0.05), while the urine creatinine and urine protein were not statistically significant as compared with those of the control group. Compared with the control group, the expression of NF-κB, NOX2, Ang II in thoracic aorta, and ROS in mesenteric artery were higher in LPS group. However, mesenteric artery NO expression was lower (P < 0.01) Conclusion LPS inflammatory stimulation during pregnancy causes kidneys inflammation and increased oxidative stress in blood vessels in offspring rats.

ON THE ROLE OF TUMOR NECROSIS FACTOR-ALPHA (TNF-α) AND NUCLEAR FACTOR p-NF-κB IN THE PENTYLENETETRAZOLE KINDLING PATHOGENESIS

Determining the role of endogenous factors as markers of chronic epileptic activity allows pathogenetically justifying new approaches to treating epilepsy. The aim of the work was the immunohistochemical study of the expression of the tumor necrosis factor-alpha (TNF-α) and nuclear factor p-NF-κB in the tissue of the dorsal parts of the hippocampus in rats with kindling seizures. Kindling was produced in 15 rats by three-week i.p. pentylenetetrazole (PTZ, 35.0 mg/kg) administration. 20 control group rats were injected with 0.9% NaCl solution. The avidin-biotin-peroxidase method was used in 10 control group rats for staining. The rest ten rats composed the negative control group and stained using only secondary antibodies. The color intensity of the brain sections of the control and kindling groups was compared with the color of the brain sections of the negative control group using the "Image J" program. In rats with PTZ-kindling, the level of TNF-α was 17.86+0.83 relative units (RU) and exceeded the corresponding indicator in the control group (4.78+0.14 RU), (p&lt;0.001). The expression of p-NF-κB was 5.24+0.61 against 1.73+0.07 RU in control (p&lt;0.001). Determination of the expression of TNF-α and NF-κB in limbic structures can be used as markers of the effectiveness of experimental treatment methods for chronic epilepsy. Key words: seizures, cytokines, hypoxia, pentylenetetrazol, hippocamp

Acute oral toxicity study of polyherbal formulation: Constac granules

Abstract BACKGROUND: Constipation is a common and frequently persistent gastrointestinal motility condition. It affects the economic burden, the standard of living, social interaction, and the capacity to carry out everyday tasks. This study aimed to evaluate the acute oral toxicity of Constac granules in Wistar rats. METHODS: A total of 12 rats were randomly divided into four groups of three each. Group 1 served as the control and received only distilled water, while groups 2 and 3 received doses of 2000 mg/kg body weight of Constac granules, and group 4 received doses of 5000 mg/kg body weight of Constac granules. The rats were observed for 14 days for any signs of toxicity. Necropsy was performed on all rats at the termination of the study and gross pathology was recorded. RESULTS: The results showed no clinical signs of toxicity or mortality observed in any of the control and treatment groups rats during the 14-day observation period. The LD50 value was estimated to be greater than 5000 mg/kg body weight. CONCLUSION: The acute oral toxicity study of Constac granules in Wistar rats showed no toxicity at doses up to 5000 mg/kg body weight. These findings showed that the oral administration of the Constac granules is safe to use.

Effects of Ethanolic Extracts of Fruits of Dennettia tripetala on Liver Function of Male Albino Rats

This study evaluated the effects of ethanolic extracts of fruits of Dennettia tripetala on liver function of male albino rats. Dennettia tripetala is an endemic plant of west tropical Africa that is known as pepper fruit. The fruit is known for its numerous medicinal properties such as antioxidant, anti-inflammatory and antidiarrhea. The plant materials were air dried and made into powder, 250g was used for the crude extraction using 70% ethanol. Ninety eight male albino rats were used in this study; forty eight were used for LD50 and fifty for the main experiment. The animals were randomly distributed into 5 groups of 10 animals each. The test animals were administered single dose of ethanolic extracts of fruits of D. tripetala (200 mg/kg and 400 mg/kg) daily for twenty-one days. The animals sacrificed at end of the 21 days. Blood sample was collected via cardiac puncture for biochemical analysis, while the liver was harvested for histological examination. A very high LD50 value of 4000 mg/kg body weight was recorded in this study. The results of biochemical parameters revealed that mean values of AST ALT and ALP were elevated significantly (p&lt;0.05) in all the test groups, except ALP in group 4 which showed no significant alteration when compared to the control. Total protein, albumin and globulin showed no significant alteration in all the test groups; total and direct bilirubin decreased significantly in test groups 4 and 5, and increased in group 2, while indirect bilirubin showed no significant alteration in all the test groups. Photomicrograph of liver section of normal control rats (group 1) showed normal central vein, portal triad, sinusoids and lamella of hepatic cells as well as those in all the test groups.

Extra virgin olive oil prevents renal histopathological damage in arsenic exposed albino rats

Introduction: To study the protective effect of extra virgin olive oil (EVOO) on histopathological changes induced by arsenic in the kidneys of albino rats. Randomized control trial. November 1, 2017–November 30, 2017 at National Institute of Health. Material and Methods: Forty-five male adult albino rats were placed in three cages having 15 rats each. Distilled water was given to the rats of control Group I for 30 days. The dose of sodium arsenite given to rats was 40 mg per kg per day dissolved in drinking water for 30 days. Olive oil of 0.2 ml per day was only given to Group III rats for 30 days along with sodium arsenite. In dissection was done after 30 days and their kidneys were approached and dissected out for histological changes. Results: EVOO has ameliorated the microscopic quantitative and qualitative histological changes induced by arsenic on both kidneys of albino rats. Olive oil had significantly prevented the increase in the diameter of the proximal convoluted tubule (PCT). Discussion and Conclusion: The present study demonstrates that EVOO prevents the quantitative and qualitative histological changes caused by arsenic on kidneys which include the diameter of PCT and distal convoluted tubule, the diameter of glomeruli, width of bowman’s space, and loss of brush border.

Electron microscopic GABA evaluation in hippocampal mossy terminals of genetic absence epilepsy rats receiving kindling stimulations

Objective: The hypotheses related to the fact of epileptic mechanisms are mainly based on excitation-inhibition imbalance in central nervous system. GAERS (Genetic Absence Epilepsy Rats from Strasbourg) is a well-known animal model of absence epilepsy, and frequently used in experimental studies. In the present study, we aimed to examine possible morphological and gamma-aminobutyric acid (GABA) density changes in GAERS hippocampus after electrical kindling stimulations.&#x0D; Methods: All control and test group rats received 6 kindling stimulations. Rats were decapitated 1 h after the last stimulation. Ultrastructural GABA immunocytochemistry was used to evaluate GABA density quantitatively in mossy terminals of hippocampal CA3 region.&#x0D; Results: GABA levels were less in kindling groups compared to their controls, and in GAERS groups compared to Wistar groups; mitochondrial and dendritic spine area ratios were greater in GAERS groups compared to Wistar groups, although all these evaluations were statistically nonsignificant. Depletion of synaptic vesicles was evident in the mossy terminals of kindling groups.&#x0D; Conclusion: The reason of decreased levels of GABA found in the present study might be that GABA has been released from the synaptic pool rapidly at an early time period after the last stimulation, for compansation mechanisms. Depletion of synaptic vesicles observed in kindling groups shows that even 6 kindling stimulations have an impact of changing hippocampal morphology in trisynaptic cycle. The increased mitochondrial area in GAERS might be related to the increased mitochondrial activity. The increased dendritic spine area might be related to the increased performance of learning in GAERS. Our findings indicating that absence epilepsy and temporal lobe epilepsy have different mechanisms of epileptogenesis might be a basis for further experimental studies

Accute Toxicity Of Extract From Melinjo (Gnetum Gnemon L) Leaf With Fixed Dose Procedure Method

Melinjo leaves has several benefits as antidiabetic, antihyperglycemia, antioxidant and diuretic. In this experiment the acute toxicity test of ethanol extract of melinjo (Gnetum gnemon L). leaves on male white rats of wistar strain with fixed dose procedure method. The result of the screening of the phytochemical showed that the leaves of beluntas contained alcaloid, flavonoid, saponin, fenolic, and tanin. Ethanol extract of melinjo leaves was obtained by maceration using ethanol 70%. From the observational study, the dose 2000 mg/kg BW was chosen as starting dose for the main study. Experimental animal subject used in the main study were 10 male white rats of wistar which were divided into 2 group, normal group and treatment group with a dose of 2000 mg/kgBW. The result showed that there were no death or toxicity sign in all experimental animals. During the 14 days observation period, there were no significant changes in body weight of rats either in normal control group and treatmen group with a dose of 2000 mg/kg BW (p&gt;0.05). Melinjo leaves had no effect on the macroscopic organs of liver, kidneys and heart of the test animals (p&gt;0.05). The average level of biochemical parameters in the normal group is SGOT 54.13±11.428 U/L, SGPT 47.476±21.655 U/L, creatinine 0.632±0.199 mg/dL, and urea 14.46±2.267 mg/dL, while the 2000 mg/kg BW dose group is SGOT 60.192±14.198 U/L, SGPT 55.968±22.998 U/L, creatinine 0.756±0.204 mg/dL, and urea 16±2.561 mg/dL. It showed that the ethanol extract of melinjo leaves with a dose of 2000 mg/kgBW are the category of practically non-toxic.

Nelsonia canescens (Acanthaceae) aqueous extract and partitioned fractions ameliorates type-2 diabetes in alloxan-induced diabetic rats

BackgroundDiabetes mellitus is a metabolic disorder that affects the body’s ability to produce or use insulin. There is a continuous rise of this disease particularly in developing countries due to changes in life style and poverty among the people. In this study, antidiabetic activities of aqueous extract of Nelsonia canescens and its partitioned fractions in alloxan-induced diabetic rats were evaluated. Male albino rats were divided into 9 groups (diabetic and non-diabetic) of 5 rats each. Diabetes was induced by single intraperitoneal administration of alloxan (90 mg/kgbwt). The experimental design consists of a diabetic control group (untreated), a normal control group (1 mL saline), a standard diabetic drug (Glibenclamide; 5 mg/kgbwt), two doses (50 and 300 mg/kgbwt) of aqueous extract, ethyl acetate and methanol fractions of Nelsonia canescens were orally administered for a period 21 days. Blood glucose of the rats was monitored at 3-days intervals while biochemical and in vivo antioxidant assays of serum and liver were determined after 21 days.ResultsThe hypoglycemic effect of the extract observed was in a dose dependent manner with a significant reduction (p < 0.05) of blood glucose in ethylaceate fraction > aqueous extract > methanol fraction compared with the diabetic control group. A significant difference (p < 0.05) in lipid profiles and serum enzyme activity of rats in the diabetic control group was observed compared with the extract and fraction treated groups.ConclusionThe results suggest that the aqueous extract and fractions of N. canescens showed hypoglycemic and hypolipidemic potentials and significantly (p < 0.05) reduced the progression of oxidative stress.

Efficiency of feed additive based on clinoptilolite in experimental combined mycotoxicosis in laboratory animals

In the course of our work, an experiment was conducted on the experimental reproduction of mixed chronic mycotoxicosis on mature white rats with a live weight of 115-125 g. For the experiment, 40 animals of the same age, weight, physiological state, and both sexes were selected. The rats were divided into 4 groups of 10 animals each. When modeling combined chronic mycotoxicosis, we used feeds naturally contaminated with fungal toxins at concentrations below the maximum allowable levels. Toxicity criteria were: general condition, mortality, change in body weight, change in behavior, morphological and biochemical parameters of blood. Animals of the 1st group received feed contaminated with toxins with a feed additive of 1 %; the 2nd group received feed contaminated with toxins with a feed additive of 2 %, the 3rd control group received feed contaminated with toxins; animals of the 4th control group received food without mycotoxins. The first signs of toxicosis in animals appeared on the 15th day of feeding toxic feed in the 3rd control group (OP + mycotoxins). In rats of the 3rd control group, deviations in behavior were noted, manifested by fearfulness and a temporary violation of movement coordination. In the same group, there was a disturbance in the activity of the gastrointestinal tract, disheveledness and loss of luster of the coat, and an increase in water consumption. Deterioration of feed intake and even refusal of it was noted in 3 animals of the 3rd control group (OP + mycotoxins) on the 14th day of feeding. The biochemical blood tests carried out on the 15th and 30th days of the experiment showed that in animals of the 3rd control group, the amount of total protein, cholesterol decreases, in comparison with the 4th control group, which received the main diet without mycotoxins. Such changes are observed in digestive disorders, parenchymal liver lesions, protein and lipid metabolic disorders. It was found that the feed additive, when introduced into the diet in an amount of 2 %, showed the greatest efficiency.

Pretreating mesenchymal stem cells with IL-6 regulates the inflammatory response of DSS-induced ulcerative colitis in rats

The immunomodulatory properties of mesenchymal stem cells (MSCs) have been broadly investigated in research on inflammatory diseases including ulcerative colitis. Treating MSCs with an inflammatory stimulus before transplantation is an adaptive strategy that helps MSCs survive in areas of inflammation and promotes the regulation of local immune responses. This study aimed to examine the effects of pretreating bone marrow MSCs (BMSCs) with Interleukin-6 (IL-6) on attenuation of dextran sulfate sodium (DSS)-induced ulcerative colitis in rats. Experimental ulcerative colitis was induced in Wistar rats by administering 2% DSS in their water for 7 days and normal water for the next 3 days. The experimental group received 1 × 106/0.4 ml of BMSCs that were treated with IL-6 for 24 h. Histological changes, colon length, and disease activity index were compared among groups, and the levels of TNF-α, IL-6, and IL-1β in homogenate supernatants were evaluated using ELISA. IL-6-pretreated BMSCs significantly reduced the colonic damage score. The colon length shortened by 6.1 ± 0.14 cm for the rats that received IL-6-pretreated BMSCs, whereas the control group rats' value was 3.8 ± 0.14 cm on the 14th day. The levels of pro-inflammatory cytokines were significantly decreased in the colons of the IL-6-pretreated BMSCs group compared with those of the control group (p < 0.05). This study revealed that IL-6-pretreated BMSCs ameliorated DSS-induced colitis via local anti-inflammatory action and suggested that IL-6-pretreated BMSCs are a promising therapeutic agent for ulcerative colitis treatment.

Evaluation of clinical, histology, TNF-α, and collagen expressions on oral ulcer in rats after treatment with areca nut and chrysanthemum oral gel

Background: Areca nut (Areca catechu Linn.) is the seed of the fruit of the oriental palm that is commonly used among Southeast Asian communities. Chrysanthemum (Dendrathema grandiflora) is a flowering plant originating from East Asia and dominantly grows in China. Both of these plants have strong antioxidant activities. To investigate the mechanism of their wound healing activities, we prepared areca nut and chrysanthemum polyethylene oral gel and performed several in vivo assays using Sprague–Dawley rats. Methods: Sprague–Dawley rats were divided into six groups: Negative control group (rats with base gel treatment), positive control group (rats treated with triamcinolone acetonide), F1 (treatment with 20% areca nut:80% chrysanthemum), F2 (treatment with 50% areca nut:50% chrysanthemum), F3 (treatment with 80% areca nut:20% chrysanthemum), and normal group (without ulcer and treatment). Traumatic ulcers were performed on the buccal mucosa of all experimental animals that received topical oral gel and triamcinolone acetonide twice a day for seven days. The clinical and histological characteristics were analyzed and scored. Results: During the six days, the ulcerated area receded linearly over time and was completely cicatrized in F2, F3, and the positive control group (Dependent t-test, p&lt;0.05). There were significant increases in body weight in F2 and positive control groups. There were no significant differences between groups in histology examination (Kruskal Wallis test, p&lt;0.05). The moderate score of TNF-α levels was seen in F2 and positive control groups (ANOVA/Tukey test, p&lt;0.05). In the collagenases assay, a high concentration of areca nut (F3) induced the abundance of collagen during the ulcer healing process. Conclusions: The combination of areca nut and chrysanthemum extract in the oral gel can optimize the healing of traumatic oral ulcers in Sprague-Dawley rats through the increase of TNF-α and collagen deposition.

Achyranthes Aspera Root Extract Attenuates the Damage Caused by High Fat Diet and Streptozotocin-Induced Diabetes in Wistar Rats

Achyranthes aspera is an herb found in the Indian subcontinent and has been used for decades because of its traditional anti-diabetic and antiobesity properties. Since studies are not reported on Achyranthes aspera protection in high-fat diet and diabetes-induced animal models, our aim was to investigate the protective effect of root extract of Achyranthesaspera on the combined effect of high-fat diet and streptozotocin-induced biochemical changes in Wistar rats. To achieve this aim, the objectives of the study were to estimate the body weight, serum glucose, insulin, Glycated hemoglobin(HbA1c), Homeostatic Model Assessment of Insulin Resistance (HOMA- IR), serum lipid profile, and liver glycogen among the Wistar rats which were divided into 6 groups having 6 rats in each group. Group 1 - control rats Group 2 - diabetic group rats, Group 3 – HFD diabetic rats and the rats from groups 4 to 6 were given a high-fat diet+ Streptozotocin 40 mg/kg + a low dose of root extract of Achyranthes aspera (250 mg/kg), high dose of root extract of Achyranthe saspera (500 mg/kg), metformin (250 mg/kg), respectively treated for 35 days. Body weight followed by serum glucose, insulin, HBA1C, HOMA- IR, serum lipid profile, and liver glycogen were evaluated. HFD + Diabetic group showed an elevated level of lipid profile and alterations in serum glucose, HbA1C, HOMAIR, Insulin and liver glycogen. Administration of Metformin and hydro alcoholic root extract of Achyranthes aspera500 mg/kg to HFD + diabetic group protected the biochemical alterations and serum insulin, glucose levels, HbA1c, HOMA- IR, liver glycogen. In comparison with HFD group, there was 1.76,2.02, and,1.3-fold decrease in Glucose levels in HFD+STZ+ Metformin, HFD+STZ+HEAA250, and HFD+STZ+HEAA500 groups respectively. In comparison with HFD group, there was a 1.38,1.15 and,1.59-fold increase in Insulin levels in HFD+STZ+ Metformin, HFD+STZ+HEAA250, and HFD+STZ+HEAA500 groups respectively. The HbA1C levels (mean±SEM) of control, STZ, HFD+STZ, HFD+STZ+ Metformin, HFD+STZ+HEAA250, and HFD+STZ+HEAA500 were 7.36±0.22,10.18±0.74,8.94±0.33,8.10±0.22,8.94±0.22 and 7.0±0.41 respectively. The HOMA-IR levels (mean±SEM) of control, STZ, HFD+STZ, HFD+STZ+ Metformin, HFD+STZ+HEAA250, andHFD+STZ+HEAA500were 2.08±0.12,3.99±0.85,17.05±3.53,1.61±0.28,2.46±0.74 and 2.01±0.20 respectively The study findings reveal the protective role of Achyranthesaspera on high-fat diet and a diabetic induced biochemical changes.

Lipid composition and oxidative changes in diabetes and alcoholic diabetes rats

BackgroundThe investigating of study was expected to the lipid composition of diabetes and alcoholic diabetes in plasma and erythrocyte membrane biochemical profile in rats. Diabetic male Wistar Streptozotocin (STZ)-induced rats were used experimental models. Control rats (C) were maintained group I, received glucose (i.e., caloric equivalent), diabetic induced rats (STZ) group II, and group III alcoholic treated and IV diabetic and alcoholic treated rats which received 20 % (v/v) alcohol in water, administered through stomach tube (5 g/kg body weight/day). ResultsSTZ-induced diabetic hepatic damage in rats was observed which leads to the increased plasma lipid peroxidation, nitrate and nitrite levels. Diabetic and administration alcohol rats also suggestively lesser the activities of antioxidants, glutathione peroxidase, glutathione S-transferase, superoxide dismutase, catalase and reduced glutathione when related with control rats (group I). Plasma enzymes are normal levels and renovate the enzymic and non-enzymatic antioxidants level in experimental groups. ConclusionThe present data point out that the nitric oxide scavenging levels increased and may possibly protect and adjacent to oxidative stress and free radicals in diabetic hepatopathy rats and histopathological studies were identified in regular hepatic cortex of Group II, III and IV of animals with diabetes, alcoholic and alcohol-induced diabetes rats.

Intervention effects of erythropoietin derived peptide (HBSP) on renal injury induced by acute skeletal muscle strain in rats

Objective: To investigate the protective effects of erythropoietin derived peptide, also known as spiral B surface peptide (HBSP), on kidney and aggregated proteins (Agrin) levels in acute skeletal muscle strain rats. Methods: Forty SPF grade SD male rats were randomly divided into control group, injury group, HBSP group and EPO group, with 10 rats in each group. Acute skeletal muscle strain animal models were established except the control group. After successful modeling, the rats in HBSP group and EPO group were intraperitoneally injected with 60 μg/kg HBSP and 5 000 U/kg recombinant human erythropoietin (rhEPO), and the rats in the control group and the injured group were intraperitoneally injected with 0.9% normal saline. Renal function was monitored with relevant kits; Hematoxylin-eosin staining was used to observe the pathological morphology of kidney tissue and skeletal muscle strain tissue. The apoptosis rate of renal tissue cells was detected by in situ terminal transferase labeling (TUNEL). Western blot and quantitative polymerase chain reaction (Q-PCR) were used to determine the expressions of Agrin and muscular-specific kinase (MuSK) in the injured skeletal muscle of rats in each group. Results: Compared with the control group, the renal function indexes serum creatinine (Cr), urea nitrogen (BUN) and 24 h urinary protein (UP24) levels of rats in injured group were increased (P＜ 0.05), but the levels of BUN, Cr and UP24 of rats in HBSP group were decreased (P＜0.05). Compared with HBSP group, there were no significant differences in the above indexes in EPO group (P＞0.05). In the control group, the muscle fiber structure was intact, the shape and structure of the fiber bundles were normal, and there was no infiltration of red blood cells and inflammatory cells in the interstitium, and no fibrohyperplasia. In the injured group, the muscle tissue showed sparse and irregular arrangement, and the interstitial widened with a large number of inflammatory cells and red blood cell infiltration. Erythrocytes and inflammatory cells were reduced in HBSP group and EPO group, and the transverse and longitudinal lines of muscle were clear. The glomerular structure of the rats in the fibrohyperplasia control group was intact and no lesions were observed. In the injured group, glomerular hypertrophy and significant matrix hyperplasia were observed, as well as the expansion of renal cysts with vacuolar and significant inflammatory infiltration were observed, and the inflammatory infiltration was reduced in the HBSP and EPO groups. Glomerular hypertrophy and hyperplasia were alleviated. The apoptosis rates of kidney cells in control group, injured group, HBSP group and EPO group were (4.05±0.51) %, (26.30±2.05) %, (14.28±1.62) % and (16.03±1.77) %, respectively, and there were significant differences among these groups (P＜0.05). Compared with control group, the levels of Agrin and MuSK in skeletal muscle pulled tissue were significantly decreased (P＜0.05), and those of HBSP group and EPO group were significantly increased compared with injured group (P＜0.05), but there was no significant difference between HBSP group and EPO group (P＞0.05). Conclusion: Erythropoietin derived peptide (HBSP) has obvious intervention effects on renal function injury in rats with acute skeletal muscle strain, and its mechanisms may be related to reducing the apoptosis rate of renal tissue cells and activating Agrin and MuSK expression.

Effects of panax notoginseng saponins on pulmonary vascular remodeling in rats with pulmonary hypertension and its mechanisms

Objective: To investigate the effects of panax notoginseng saponins (PNS) on pulmonary vascular remodeling and SIRT1/FOXO3a/p27 pathway in pulmonary arterial hypertension (PAH) rats. Methods: Male SD rats weighing 200~250g were randomly divided into control group, monocrotaline group (MCT) and monocrotaline + panax notoginseng saponins group (MCT+PNS), with 10 rats in each group. The rats in control group were injected intraperitoneally with normal saline 3 ml/kg on the first day, then injected intraperitoneally with normal saline 2.5 ml/kg every day. The rats in MCT group were injected intraperitoneally with MCT 60 mg/kg on the first day, followed by daily injection of normal saline 2.5 ml/kg. In MCT+PNS group, 60 mg/kg MCT was injected intraperitoneally on the first day, and 50 mg/kg PNS was injected intraperitoneally every day. The above models were fed conventionally for 4 weeks. After the modeling was completed, the mean pulmonary artery pressure (mPAP) and right ventricular systolic pressure (RVSP) of rats in each group were detected by right heart catheter method, weighed and calculated right ventricular hypertrophy index (RVHI), and the pulmonary vascular structure and morphological changes were observed by HE and Masson staining. The protein and gene expressions of SIRT1, FOXO3a, p27, PCNA and Caspase-3 were detected by qPCR and Western blot. Results: Compared with control group, mPAP, RVSP and RVHI in MCT group were increased significantly (P＜0.01), pulmonary vessels were thickened significantly and collagen fibers were increased, protein and gene expressions of SIRT1, FOXO3a, p27 and Caspase-3 were decreased (P＜0.05 or P＜0.01). The protein and gene expressions of PCNA were increased (P＜0.05). Compared with MCT group, the levels of mPAP, RVSP and RVHI in MCT+PNS group were decreased significantly (P＜0.05 or P＜0.01), pulmonary vascular thickening was alleviated and collagen fibers were reduced. The protein and gene expressions of SIRT1, FOXO3a, p27 and Caspase-3 were increased (P＜0.05 or P＜0.01), while the protein and gene expressions of PCNA were decreased (P＜0.05 or P＜0.01). Conclusion: Panax notoginseng saponins can relieve pulmonary vascular remodeling in rats with pulmonary hypertension by activating SIRT1/FOXO3a/p27 pathway.

Effect of extracellular signal-regulated kinase inhibitor on calpain in rat cerebral cortex after cardiopulmonary resuscitation

To explore the effect of extracellular signal-regulated kinase (ERK) inhibitor PD98059 on calpain-related proteins in the brain, and to understand the pathophysiological changes of calpain in cerebral ischemia/reperfusion injury (CIRI). Forty-two rats were divided into sham operation (Sham) group (n = 6), model group (n = 12), dimethyl sulfoxide (DMSO) control group (n = 12), and PD98059 group (n = 12) by random number table. The rat model of CIRI induced by cardiac arrest-cardiopulmonary resuscitation (CA-CPR) was reproduced by transesophageal electrical stimulation to induce ventricular fibrillation. In the Sham group, only the basic operations such as anesthesia, tracheal intubation, and arteriovenous catheterization were performed without CA-CPR. The rats in the DMSO control group and PD98059 group were injected with DMSO or PD98059 0.30 mg/kg via femoral vein, respectively, 30 minutes after the restoration of spontaneous circulation (ROSC), and rats in the Sham group and model group were given the same amount of normal saline. The duration of CPR, 24-hour survival rate and neurological deficit score (NDS) after ROSC were recorded. Hematoxylin-eosin (HE) staining and Nissl staining were used to observe the pathological changes of the cerebral cortex. The expressions of phosphorylated ERK (p-ERK), ERK, calpastatin, calpain-1, and calpain-2 were detected by Western blotting. The co-expression of p-ERK and calpain-2 was detected by double immunofluorescence. There were no significant differences in the duration of CPR and 24-hour survival rate among all groups. In the model group, the nuclei of the cerebral cortex were obviously deformed and pyknotic, cells vacuoles and tissues were arranged disorderly, Nissl corpuscles were significantly reduced, NDS scores were also significantly reduced, level of ERK phosphorylation was increased, and calpain-2 protein was significantly up-regulated compared with the Sham group. There was no significant difference in the above parameters between the DMSO control group and the model group. After intervention with PD98059, the pathological injury of brain tissue was significantly improved, Nissl corpuscles were significantly increased, the NDS score was significantly higher than that in the model group [75.0 (72.0, 78.0) vs. 70.0 (65.0, 72.0), P < 0.05], the level of ERK phosphorylation and calpain-2 protein expression were significantly lower than those in the model group [p-ERK (p-ERK/ERK): 0.65±0.12 vs. 0.92±0.05, calpain-2 protein (calpain-2/GAPDH): 0.73±0.10 vs. 1.07±0.14, both P < 0.05], while there was no significant difference in the expressions of calpastatin and calpain-1 in the cerebral cortex among all the groups. Double immunofluorescence staining showed that p-ERK and calpain-2 were co-expressed in cytosol and nucleus, and the co-expression rate of p-ERK and calpain-2 in the model group was significantly higher than that in the Sham group [(38.6±4.3)% vs. (9.2±3.5)%, P < 0.05], while it was significantly lowered in the PD98059 group compared with the model group [(18.2±7.0)% vs. (38.6±4.3)%, P < 0.05]. ERK together with calpain-2 participated in CIRI induced by CA-CPR. PD98059 inhibited the expression of calpain-2 and ERK phosphorylation. Therefore, ERK/calpain-2 may be a novel therapeutic target for CIRI.

Resistant dextrin protects against pathological bone loss in ovariectomized rats and inhibits RANKL-induced osteoclastogenesis.

Osteoporosis is a common disease in postmenopausal women characterized by systemic bone mass loss, microstructure fragility and increased incidence of fractures. Resistant dextrin (RD) is a soluble fiber with beneficial metabolic effects. However, the beneficial effect of RD in osteoporosis remains to be determined. In this study, we investigated the effect of dietary RD supplement on osteoporosis in ovariectomized (OVX) rats. Both the control (sham) and OVX group rats were gavaged with RD (10 g/kg/d) or equal amount of saline for 12 weeks, and histological and biomechanical analyses were conducted to evaluate bone microstructure and strength. Furthermore, we also evaluated the effects of RD on osteoclastogenesis in bone marrow macrophages (BMMs) by detecting the expression of osteoclast-related genes using qRT-PCR and Western blot analysis. The results showed that in OVX rats the bone strength and microstructure characteristics were significantly improved with RD supplement for 12 weeks. Additionally, the mRNA and protein expression of osteoclast markers, such as CTSK, NF-κB and NFATC1, were significantly down-regulated in BMMs isolated from RD supplement group. RD also suppressed RANKL-induced osteoclastogenesis in BMMs. These findings suggest that RD ameliorates osteoporosis in OVX rats by inhibiting osteoclast differentiation. RD suppresses RANKL-induced osteoclastogenesis possibly through modulating Akt and NF-κB signaling pathways. These data indicate that a dietary supplement of RD might serve as an intervention strategy for menopausal osteoporosis.