Healthy Control Rats Research Articles

Effects of Crystal Derived from Stevia rebaudiana Leaves on Alloxan Induced Type-1 Diabetic Mice

The study was undertaken to investigate the antidiabetic effects of crystals derived from Stevia rebaudiana leaves in normal rats and alloxan induced diabetic mice. Normal rats were used to evaluate whether the stevia crystal have any adverse effects on body weight and blood glucose of healthy rats. The rats were divided into two groups (n=5); healthy control rats and stevia treated healthy rats (1 g/kg/day orally). To evaluate the antidiabetic effects of stevia crystal, mice were divided into five equal (n=5) groups; healthy control, diabetic control, stevia crystal @ 250 and 500 mg/kg bwt, and Amaryl® @ 800 µg/kg daily to compare the efficacy. Alloxan monohydrate was injected in mice at a dose rate of 120 mg/kg intraperitoneally for diabetic induction. Result of the present study showed that stevia crystal at a high daily dose did not have any significant effect on body weight but it reduced blood glucose level non-significantly in healthy animal. The antidiabetic effects of stevia crystal was evaluated based on glucose lowering capacity, improvement of body weight loss, changes in lipid profile, and renal and pancreas protective capacity. Treatment with stevia crystal @500 mg/kg improved body weight loss and reduced blood glucose level significantly (P<0.01) in diabetic animal. Stevia @500 mg/kg decreased cholesterol level non-significantly whereas it reduce the triglyceride level significantly (P<0.01). In histopathology study, stevia crystal showed renal and pancreas protective effect with slightly restoration of structural damage in both organs. Based on present studies data it may conclude that crystalline compounds derived from Stevia rebaudiana leaves may have antidiabetic properties that need further characterization

Characterization of liver changes in ZSF1 rats, an animal model of metabolic syndrome.

The non-alcoholic fatty liver disease is the hepatic counterpart of the metabolic syndrome. ZSF1 rats are a metabolic syndrome animal model in which liver changes have not been described yet. The characterization of liver histological and innate immunity changes in ZSF1 rats. Five groups of rats were included (n = 7 each group): healthy Wistar-Kyoto control rats (Ctrl), hypertensive ZSF1 lean (Ln), ZSF1 obese rats with a normal diet (Ob), ZSF1 obese rates with a high-fat diet (Ob-HFD), and ZSF1 obese rats with low-intensity exercise training (Ob-Ex). The animals were sacrificed at 20 weeks of age, their livers were collected for: a) measurements of the area of steatosis, fibrosis and inflammation (histomorphological analysis); and b) innate immunity (toll-like receptor [TLR] 2, TLR4, peroxisome proliferator-activated receptor γ [PPARγ], toll interacting protein [TOLLIP]) and inflammatory marker (tumor necrosis factor-alpha [TNFα], interleukin 1 [IL-1]) expression analysis by real-time PCR. Ob, Ob-HFD and Ob-Ex were significantly heavier than Ln and Ctrl animals. Ob, Ob-HFD and Ob-Ex animals had impaired glucose tolerance and insulin resistance. ZSF1 Ob, Ob-HFD and Ob-Ex presented a higher degree of steatosis (3,5x; p < 0.05) than Ctrl or ZSF1 Ln rats. Steatohepatitis and fibrosis were not observed in any of the groups. No differences in expression were observed between Ctrl, Ln and Ob animals (except for the significantly higher expression of TOLLIP observed in the Ob vs Ln comparison). Ob-HFD and Ob-Ex rats showed increased expression of PPARγ and TOLLIP as compared to other groups. However, both groups also showed increased expression of TLR2 and TLR4. Nevertheless, this did not translate into a differential expression of TNFα or IL-1 in any of the groups. The ZSF1 model is associated with liver steatosis but not with steatohepatitis or a significantly increased expression of innate immunity or inflammation markers.

MALDI-LTQ-Orbitrap mass spectrometry imaging for lipidomic analysis in kidney under cisplatin chemotherapy

Imaging techniques for mapping molecular distributions in tissue sections can reveal valuable information on biomolecules involved in relevant biochemical processes. A method has been developed for comprehensive, reproducible and sensitive lipid imaging by matrix-assisted laser/desorption ionization-LTQ-Orbitrap mass spectrometry in kidney sections, showing the benefits of exact mass determination. Matrix deposition parameters for positive and negative lipid ion imaging using different matrices such as 2,5-dihydroxybenzoic acid (DHB), 9-aminoacridine (9-AA) or α-cyano-4-hydroxycinnamic acid (CHCA) have been optimized for the broadest detection and identification of renal lipids. The combination of 9-AA and DHB was found as the most suitable for negative and positive ion mode lipid imaging, respectively. Lipid mapping and related identification strategies and limitations have also been discussed. Production of 100-µm resolution images was proved to be enough for discerning lipid distribution in kidney substructures. Imaging reproducibility was assessed on parallel kidney slices with time. This method has been applied to the lipidomics analysis on kidney sections from rats treated with the antitumor drug cisplatin and compared to healthy control rats. Up to 66 different renal lipids out of 450 extracted ion images (mainly phospholipid species, in addition to sulfatides and cholesterol sulfate) have been found and identified showing a modified distribution pattern due to cisplatin-induced nephrotoxicity. These lipid species reflect either topographic, signaling or structural processes in damaged kidney and could potentially be used for nephrotoxicity assessment or as therapeutic targets. This is, to our knowledge, the first imaging lipidomics study for nephrotoxicity assessment of cisplatin chemotherapy.

Injection of hTERT-Transduced Endothelial Progenitor Cells Promotes Beneficial Aortic Changes in a High-Fat Dietary Model of Early Atherosclerosis

Objectives: Cultured endothelial progenitor cells (EPCs) display troubling issues that adversely affect their applicability to endothelial regeneration. We hypothesized that transduction of the human telomerase catalytic subunit (hTERT) gene would enhance EPC function in treating dietary-induced early atherosclerosis (AS). Methods: A dietary-induced early AS model was successfully constructed in 90 healthy male rats, while 30 healthy control (HC) rats were normally fed. Four experimental groups were constructed: an untreated HC group; an untreated AS group injected with PBS; a null EPC AS group injected with null vector-transduced EPCs, and an hTERT EPC AS group injected with hTERT-transduced EPCs. Two months postinjection, abdominal aortas were extracted to validate EPC integration and comparatively assess mRNA and protein expression of the early atherosclerotic markers VCAM-1, ICAM-1, LFA-1, Mac-1, CD44, MCP-1, endothelial nitric oxide synthase (eNOS), and apolipoprotein E. Results: In vitro, hTERT transduction of EPCs resulted in a significantly superior proliferative capacity as well as significantly higher NO, iNOS, and LDH secretory capacity. In vivo injection of hTERT-transduced EPCs produced significant reductions in CD44 and MCP-1 expression as well as a significant increase in eNOS expression relative to injection with null vector-transduced EPCs (all p < 0.05). Conclusion: hTERT-transduced human EPCs may be useful in treating dietary-induced early AS.

La electroacupuntura disminuye la 5-HT, el CGRP y aumenta el NPY en el eje cerebro-intestino en dos modelos de rata con síndrome de colon irritable, con predominio de diarrea (D-SCI)

BackgroundTo examine whether electro-acupuncture (EA) could decrease 5-hydroxytryptamine (5-HT) andcalcitonin gene-related peptide (CGRP), and increase neuro-peptide Y (NPY) in the brain-gut axis (BGA) in D-IBSusing rat models. MethodsRats were randomly exposed to unpredictable chronic stress for 3 weeks followed by 1-hour acute restraint stress (CAS) after 7 days of rest, or daily gavage of Senna decoction (6g/kg) plus chronic restraint stress (for a duration of 2h, starting from 1h prior to the gavage) for 2 weeks (ISC). The content of 5-HT, CGRP and NPYin the distal colon, spinal cord, hypothalamus was examined at the end of the treatment. Results1. The two rat models exhibited similar characteristics, e.g., increased number of fecal pellets expelled in1h, decreased sacchar-intake, decreased CRD, elevated 5-HT, CGRP content and decreased NPY in the distal colon,spinal cord, hypothalamus (P<0.05 vs. that in healthy control rats). 2. A series of equations was developed basedon correlation regression analysis. The analysis results demonstrated that 5-HT mediates the changes in hypothalamus, spinal cord and colon. 5-HT and CGRP in spinal cord was closely correlated with general behaviorevaluation and other transmitters in BGA. Conclusion1. In comparison to 5-HT, CGRP and NPY (particularly in the spinal cord) had closer relationship withthe D-IBS symptoms induced by either stress factors or Senna decotion.2. EA treatment could restore the brain-gut axis to balanced levels.

The effect of experimental osteoporosis on bone regeneration: Part 1, histology findings.

To histologically define the healing events occurring in calvarial critical size defects (CSDs) following treatment with a collagen barrier for guided bone regeneration (GBR) and a particulate graft in healthy and osteoporotic conditions. Thirty-six 10-month-old, female, Wistar rats were used in this study. Half of them were ovariectomized (OVX) and fed with a low-calcium diet to induce an osteoporotic-like status. In each animal of both groups, two 5-mm CSDs were created, one in the centre of each parietal bone, and they were treated with a deproteinized bovine bone mineral (DBBM) particulate graft and a bi-layer collagen membrane. Six OVX and six healthy control rats were randomly euthanized at 7, 14 and 30days. One defect per animal was randomly processed for decalcified histology. Three central sections were used for qualitative histology and histomorphometric analysis. No significant difference in terms of percentage of newly formed bone was detected between the two groups at the different healing periods. However, a trend towards less bone formation and of poorer quality, expressed as reduced bone maturation, was detected in the OVX animals at 30days. According to this study, GBR with a collagen barrier and a DBBM graft can be successfully obtained also in osteoporotic-like conditions. Future studies considering longer healing periods and controlling for the confounding factors arising from the use of a particulate graft are needed to confirm these data.

Diabetes induced renal urea transport alterations assessed with 3D hyperpolarized 13 C,15 N-Urea.

In the current study, we investigated hyperpolarized urea as a possible imaging biomarker of the renal function by means of the intrarenal osmolality gradient. Hyperpolarized three-dimensional balanced steady state 13 C MRI experiments alongside kidney function parameters and quantitative polymerase chain reaction measurements was performed on two groups of rats, a streptozotocin type 1 diabetic group and a healthy control group. A significant decline in intrarenal steepness of the urea gradient was found after 4 weeks of untreated insulinopenic diabetes in agreement with an increased urea transport transcription. MRI and hyperpolarized [13 C,15 N]urea can monitor the changes in the corticomedullary urea concentration gradients in diabetic and healthy control rats. Magn Reson Med 77:1650-1655, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Peptidomic Analysis of Rat Plasma: Proteolysis in Hemorrhagic Shock.

It has been previously shown that intestinal proteases translocate into the circulation during hemorrhagic shock and contribute to proteolysis in distal organs. However, consequences of this phenomenon have not previously been investigated using high-throughput approaches. Here, a shotgun label-free quantitative proteomic approach was utilized to compare the peptidome of plasma samples from healthy and hemorrhagic shock rats to verify the possible role of uncontrolled proteolytic activity in shock. Plasma was collected from rats after hemorrhagic shock (HS) consisting of 2-h hypovolemia followed by 2-h reperfusion, and from healthy control (CTRL) rats. A new two-step enrichment method was applied to selectively extract peptides and low molecular weight proteins from plasma, and directly analyze these samples by tandem mass spectrometry. One hundred twenty-six circulating peptides were identified in CTRL and 295 in HS animals. Ninety-six peptides were present in both conditions; of these, 57 increased and 30 decreased in shock. In total, 256 peptides were increased or present only in HS confirming a general increase in proteolytic activity in shock. Analysis of the proteases that potentially generated the identified peptides suggests that the larger relative contribution to the proteolytic activity in shock is due to chymotryptic-like proteases. These results provide quantitative confirmation that extensive, system-wide proteolysis is part of the complex pathologic phenomena occurring in hemorrhagic shock.

Effect of &lt;i&gt;Phellodendron chinense&lt;/i&gt; Schneid Extract on Chronic Bacterial Prostatitis Induced by Chlamydia in Rats

Purpose: To explore the effects of Phellodendron chinense Schneid (PCS) extract on chlamydiainduced chronic bacterial prostatitis (CBP). Methods: Sixty 8-week-old male Sprague-Dawley rats were used in this study. Prostate index (PI) and prostate specific antigen (PSA) were determined after 4 weeks of oral administration of PCS extract (80, 160 or 320 mg/kg) or tetracycline (80 mg/kg) and compared to PI and PSA in untreated rats with CBP and healthy control rats (n = 10). Chronic inflammatory cell infiltrates, acinar changes, and interstitial fibrosis were evaluated by histopathological examination. In addition, pertinent inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), prostaglandin E2 (PEG2), transforming growth factor β1 (TGF-β1), and connective tissue growth factor (CTGF), were measured in prostate tissues using ELISA kits. Results: High doses of PCS (160 and 320 mg/kg) significantly decreased PI and PSA relative to model group (p < 0.01). PCS treatment also significantly reduced chronic inflammatory cell infiltrates and interstitial fibrosis in prostate tissue of CBP rats. In addition, TNF-α, IL-1β, COX-2, PEG2, TGF-β1, and CTGF decreased in PCS-treated rats compared to untreated control (p < 0.01). Conclusion: PCS extract has significant anti-inflammatory effects on chlamydia-induced CBP. Keywords: Phellodendron chinense Schneid, Bacterial prostatitis, Inflammatory factors, Morphometric analysis, Interstitial fibrosis, Prostate specific antigen, Prostate index

Effects and mechanisms of acupuncture and moxibustion on reproductive endocrine function in male rats with partial androgen deficiency.

Partial androgen deficiency of the aging male (PADAM) is characterised by a deficiency in serum androgen levels. Both electroacupuncture (EA) and mild moxibustion (MM) can raise serum testosterone levels in PADAM. We investigated the mechanisms underlying the use of EA and MM in a rodent model of PADAM. Fifty rats received cyclophosphamide injection over 5 consecutive days to induce PADAM, which was verified by comparing total testosterone (TT) and free testosterone (FT) levels with 10 non-PADAM healthy control rats (CON). Successful modelling was confirmed in 43 of 50 rats, 40 of which were randomly divided into untreated (PADAM), EA-treated (PADAM+EA), MM-treated (PADAM+MM), and androlin (AD)-treated (PADAM+AD) groups (n=10 each). EA and MM were administered at BL23 and CV4 acupuncture points for 8 weeks, and no treatment was given to rats in the PADAM and CON groups. Serum levels of luteinising hormone (LH) and follicle-stimulating hormone (FSH), mRNA expression of cytochrome P450c17 (P450c17) and 3β-hydroxysteroid dehydrogenase 1 (3β-HSD1), and protein levels of cytochrome P450 side chain cleavage (P450scc), 17β-hydroxysteroid dehydrogenase 3 (17β-HSD3) and steroidogenic factor 1 (SF-1) were evaluated after 8 weeks. Both EA and mild MM significantly increased serum TT and FT levels with MM displaying superiority. P450scc, 17β-HSD3 and SF-1 protein expression, and P450c17 and 3β-HSD1 mRNA expression, were significantly increased and serum LH and FSH levels were significantly decreased in PADAM+EA and PADAM+MM relative to PADAM rats. Moreover, serum LH and FSH levels were significantly lower and 17β-HSD3 protein expression significantly higher in PADAM+MM relative to PADAM+EA rats. EA and MM at the BL23 and CV4 acupuncture points appear to be effective treatments for PADAM, and MM displays superior efficacy to EA.

Hypolipidemic and antioxidant activity of the aqueous extract from the uneaten pulp of the fruit from Cordia dichotoma in healthy and hyperlipidemic Wistar albino rats.

Hyperlipidemia is a major risk factor for coronary heart disease. Hyperlipidemia increases the incidence of myocardial ischemia and cardiac events. This study evaluated the potential hypolipidemic and antioxidant action of the aqueous extract from the uneaten pulp of the fruit from Cordia dichotoma ("CDNP extract"). In vivo studies were performed for 10weeks on dietary hyperlipidemic and healthy Wistar albino rat models that received two dose levels of the CDNP extract (0.50 and 1.00g/kg body weight). Serum lipid profiles were determined for the experimental animals. Dietary hyperlipidemic rats were characterized by an elevated lipid profile compared to the healthy control, i.e., increased levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), and triglycerides (TG), although the level of high-density lipoprotein (HDL-C) was reduced. Levels of antioxidant enzymes, i.e., glutathione reductase (GR), glutathione peroxidase (GPx), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT), were significantly higher in the dietary hyperlipidemic rats than in the normal healthy ones, while the level of malondialdehyde (MDA) was significantly lower. Force-feeding hyperlipidemic Wistar albino rats with the CDNP extract at two doses decreased TC, LDL-C, VLDL-C, and TG to normal levels. The risk ratio, which was as high as 870 % for the hyperlipidemic rats was decreased by the treatment to levels close to that calculated for the healthy control rats. Levels of high-density lipoprotein cholesterol (HDL-C) were very low in the hyperlipidemic Wistar albino rats but increased significantly when CDNP extract was adminstered, attaining similar HDL-C levels to those of healthy control rats. Treatment with the CDNP extract also improved the levels of antioxidant enzymes (GR, GST, GPx, SOD, and CAT) in hyperlipidemic Wistar albino rats. Thus, the CDNP extract improves the lipid metabolism of healthy and hyperlipidemic Wistar albino rats and can be employed in the management of dietary hyperlipidemia.

Therapeutic efficacy of differentiated versus undifferentiated mesenchymal stem cells in experimental type I diabetes in rat

Selective MSCs differentiation protocol into pancreatic beta cells was conducted in the present study using exendin-4 and TGF-beta. Differentiated and undifferentiated MSCs were assessed in experimental type I diabetes in rats. Ninety female white albino rats were included in the study and divided equally (n=15/group) into 6 groups: healthy control, healthy control rats received acellular tissue culture medium, diabetic rats, diabetic rats received acellular tissue culture medium, diabetic rats received undifferentiated MSCs and diabetic rats received differentiated MSCs. Therapeutic efficacy of undifferentiated versus differentiated MSCs was evaluated via assessment of quantitative gene expressions of insulin1, insulin 2, Smad-2, Smad-3, PDX-1, PAX-4, neuroD. Blood glucose and insulin hormone levels were also assessed. Results showed that quantitative gene expressions of all studied genes showed significant decrease in diabetic rat groups. Use of undifferentiated and differentiated MSCs led to a significant elevation of expression levels of all genes with more superior effect with differentiated MSCs except smad-2 gene. As regards insulin hormone levels, use of either undifferentiated or differentiated MSCs led to a significant elevation of its levels with more therapeutic effect with differentiated MSCs. Blood glucose levels were significantly decreased with both undifferentiated and differentiated MSCs in comparison to diabetic groups but its levels were normalized 2 months after injection of differentiated MSCs. In conclusion, use of undifferentiated or differentiated MSCs exhibited significant therapeutic potentials in experimental type I diabetes in rats with more significant therapeutic effect with the use of differentiated MSCs.

High glucose concentration induces endothelial cell proliferation by regulating cyclin‐D2‐related miR‐98

Cyclin D2 is involved in the pathology of vascular complications of type 2 diabetes mellitus (T2DM). This study investigated the role of cyclin‐D2‐regulated miRNAs in endothelial cell proliferation of T2DM. Results showed that higher glucose concentration (4.5 g/l) significantly promoted the proliferation of rat aortic endothelial cells (RAOECs), and significantly increased the expression of cyclin D2 and phosphorylation of retinoblastoma 1 (p‐RB1) in RAOECs compared with those under low glucose concentration. The cyclin D2‐3′ untranslated region is targeted by miR‐98, as demonstrated by miRNA analysis software. Western blot also confirmed that cyclin D2 and p‐RB1 expression was regulated by miR‐98. The results indicated that miR‐98 treatment can induce RAOEC apoptosis. The suppression of RAOEC growth by miR‐98 might be related to regulation of Bcl‐2, Bax and Caspase 9 expression. Furthermore, the expression levels of miR‐98 decreased in 4.5 g/l glucose‐treated cells compared with those treated by low glucose concentration. Similarly, the expression of miR‐98 significantly decreased in aortas of established streptozotocin (STZ)‐induced diabetic rat model compared with that in control rats; but cyclin D2 and p‐RB1 levels remarkably increased in aortas of STZ‐induced diabetic rats compared with those in healthy control rats. In conclusion, this study demonstrated that high glucose concentration induces cyclin D2 up‐regulation and miR‐98 down‐regulation in the RAOECs. By regulating cyclin D2, miR‐98 can inhibit human endothelial cell growth, thereby providing novel therapeutic targets for vascular complication of T2DM.

Featured Article: Inhibition of diabetic cataract by glucose tolerance factor extracted from yeast.

Diabetes leads to many complications; among them is the development of cataract. Hyperglycemia brings to increased polyol concentration in the lens, to glycation of lens proteins, and to elevated level of ROS (Reactive Oxygen Species) causing oxidative stress. The glucose tolerance factor (GTF) was found by several groups to decrease hyperglycemia and oxidative stress both in diabetic animals and humans. The aim of our study was to explore the damages induced by high glucose to the eye lens and to assess the protective effects of GTF both in vivo and in vitro The in vivo study included control healthy rats, streptozotocin (STZ) diabetic untreated rats, and STZ diabetic rats orally treated with 15 doses of GTF. The diabetic untreated rats developed cataracts, whereas the development of cataract was totally or partially prevented in GTF treated animals. In vitro studies were done on bovine lenses incubated for 14 days. Half of the lenses were incubated in normal glucose conditions, and half in high glucose conditions (450 mg%). To one group of the normal or high glucose condition GTF was added. The optical quality of all the lenses was measured daily by an automated scanning laser system. The control lenses, whether with or without GTF addition, did not show any reduction in their quality. High glucose conditions induced optical damage to the lenses. Addition of GTF to high glucose conditions prevented this damage. High glucose conditions affected the activity of aldose reductase and sodium potassium ATPase in lens epithelial cell. Addition of GTF decreased the destructive changes induced by high glucose conditions. The amount of soluble cortical lens proteins was decreased and structural changes were detected in lenses incubated in high glucose medium. These changes could be prevented when GTF was added to high glucose medium. Our findings demonstrate the anticataractogenic potential of GTF.

Endogenous L-Carnosine Level in Diabetes Rat Cardiac Muscle.

A novel method for quantitation of cardiac muscle carnosine levels using HPLC-UV is described. In this simple and reliable method, carnosine from the rat cardiac muscle and the internal standard, thymopentin, were extracted by protein precipitation with acetonitrile. The method was linear up to 60.96 μg·mL−1 for L-carnosine. The calibration curve was linear in concentration ranges from 0.5 to 60.96 μg·mL−1. The relative standard deviations obtained for intra- and interday precision were lower than 12% and the recoveries were higher than 90% for both carnosine and internal standard. We successfully applied this method to the analysis of endogenous carnosine in cardiac muscle of the diabetes rats and healthy control rats. The concentration of carnosine was significantly lower in the diabetes rats group, compared to that in the healthy control rats. These results support the usefulness of this method as a means of quantitating carnosine and illustrate the important role of L-carnosine in cardiac muscle.

Qinggan Huoxue Recipe suppresses epithelial-to-mesenchymal transition in alcoholic liver fibrosis through TGF-β1/Smad signaling pathway.

To investigate the mechanism by which Qinggan Huoxue Recipe (QGHXR) inhibits epithelial-to-mesenchymal transition (EMT) in rats with alcoholic liver fibrosis (ALF). A total of 75 male SD rats were used to induce ALF. Serum biochemical indicators, including alanine aminotransferase, aspartate aminotransferase, laminin and hyaluronidase, were measured. Liver histopathological changes were evaluated using hematoxylin-eosin and Sirius red staining. EMT was examined by analyzing the expression of the epithelial marker E-cadherin and the mesenchymal markers vimentin and fibronectin using RT-PCR and Western blot. The inhibitory effect of QGHXR on EMT markers, as well as its effect on molecules associated with the transforming growth factor (TGF)-β1/Smad signaling pathway, including TGF-β1, Smad3, snail, occludin, ZO-1 and claudin, was also examined. Compared with normal control rats, ALF rats exhibited a decrease in E-cadherin levels (mRNA: ALF 0.16 ± 0.05 vs control 1.00 ± 0.08; protein: ALF 0.09 ± 0.05 vs control 0.70 ± 0.17, P < 0.01) and an increase in vimentin and fibronectin levels (mRNA: 11.43 ± 0.39 vs 1.00 ± 0.19 and 9.91 ± 0.34 vs 1.00 ± 0.44, respectively, P < 0.01; protein: 1.13 ± 0.42 vs 0.09 ± 0.03 and 1.16 ± 0.43 vs 0.09 ± 0.00, respectively, P < 0.01). This indicates that EMT occurred in ALF rats. In addition, the TGF-β1/Smad signaling pathway was activated in ALF rats, as evidenced by the increase in TGF-β1 and snail levels (mRNA: 1.76 ± 0.12 vs 1.00 ± 0.05 and 6.98 ± 0.41 vs 1.00 ± 0.10, respectively, P < 0.01; protein: 1.43 ± 0.05 vs 0.12 ± 0.03 and 1.07 ± 0.29 vs 0.07 ± 0.02, respectively, P < 0.01) and the decrease in Smad3 levels (mRNA: 0.05 ± 0.01 vs 1.00 ± 0.12, P < 0.01; protein: 0.06 ± 0.05 vs 0.89 ± 0.12, P < 0.01). Furthermore, levels of the tight junction markers occludin, ZO-1 and claudin decreased in ALF rats compared with healthy control rats (mRNA: 0.60 ± 0.09 vs 1.00 ± 0.12, 0.11 ± 0.00 vs 1.00 ± 0.12 and 0.60 ± 0.01 vs 1.00 ± 0.08, respectively, P < 0.01; protein: 0.05 ± 0.01 vs 0.87 ± 0.40, 0.09 ± 0.05 vs 0.89 ± 0.18 and 0.04 ± 0.03 vs 0.95 ± 0.21, respectively, P < 0.01). In ALF rats treated with QGHXR, E-cadherin levels increased (mRNA: QGHXR 0.67 ± 0.04 vs ALF model 0.16 ± 0.05, P < 0.01; protein: QGHXR 0.66 ± 0.21 vs ALF model 0.09 ± 0.05, P < 0.01), and vimentin and fibronectin levels decreased (mRNA: 6.57 ± 1.05 vs 11.43 ± 0.39 and 1.45 ± 1.51 vs 9.91 ± 0.34, respectively, P < 0.01; protein: 0.09 ± 0.03 vs 1.13 ± 0.42 and 0.10 ± 0.01 vs 1.16 ± 0.43, respectively, P < 0.01). In addition, QGHXR inhibited the expression of TGF-β1 and increased the expression of Smad3 (mRNA: 1.03 ± 0.11 vs 1.76 ± 0.12, 0.70 ± 0.10 vs 0.05 ± 0.01, respectively, P < 0.05 and P < 0.01; protein: 0.12 ± 0.03 vs 1.43 ± 0.05 and 0.88 ± 0.20 vs 0.06 ± 0.05, respectively, P < 0.01). QGHXR treatment also reduced the levels of the EMT-inducing transcription factor snail (mRNA: 2.28 ± 0.33 vs 6.98 ± 0.41, P < 0.01; protein: 0.08 ± 0.02 vs 1.07 ± 0.29, P < 0.01) and increased the occludin, ZO-1 and claudin levels (mRNA: 0.73 ± 0.05 vs 0.60 ± 0.09, 0.57 ± 0.04 vs 0.11 ± 0.00 and 0.68 ± 0.03 vs 0.60 ± 0.01, respectively, P < 0.01, P < 0.01 and P < 0.05; protein: 0.92 ± 0.50 vs 0.05 ± 0.01, 0.94 ± 0.22 vs 0.09 ± 0.05 and 0.94 ± 0.29 vs 0.04 ± 0.03, respectively, P < 0.01). The effects of QGR and HXR on the TGF-β1/Smad signaling pathway were similar to that of QGHXR; however, the QGR- and HXR-induced changes in vimentin mRNA levels, the QGR-induced changes in fibronectin mRNA levels and the HXR-induced changes in snail and TGF-β1 mRNA levels were not significant. Qinggan Huoxue Recipe inhibits EMT in ALF rats by modulating the TGF-β1/Smad signaling pathway, suggesting that the mechanism underlying the amelioration of ALF induced by QGHXR is associated with this pathway.

Assessment of MMP-1, MMP-8 and TIMP-2 in experimental periodontitis treated with kaempferol.

PurposeThe objective of this study was to investigate the effect of a dietary flavonoid, kaempferol, which has been shown to possess antiallergic, anti-inflammatory, anticarcinogenic, and antioxidant activities on the periodontium by histomorphometric analysis and on gingival tissue matrix metalloproteinase-1 (MMP-1), MMP-8, and tissue inhibitor of metalloproteinase-2 (TIMP-2) by biochemical analysis of rats after experimental periodontitis induction.MethodsSixty Wistar rats were randomly divided into six groups of ten rats each, and silk ligatures were placed around the cervical area of the mandibular first molars for 15 days, except in the healthy control rats. In the experimental periodontitis groups, systemic kaempferol (10 mg/kg/2d) and saline were administered by oral gavage at two different periods (with and without the presence of dental biofilm) to all rats except for the ten non-medicated rats. Alveolar bone area, alveolar bone level, and attachment level were determined by histomorphometric analysis, and gingival tissue levels of MMP-1, MMP-8, and TIMP-2 were detected by biochemical analysis.ResultsSignificantly greater bone area and significantly less alveolar bone and attachment loss were observed in the kaempferol application groups compared to the control groups (P<0.05). In addition, gingival tissue MMP-1 and -8 levels were significantly lower in the kaempferol application groups compared to the control groups and the periodontitis group (P<0.001). There were no statistically significant differences in TIMP-2 levels between the kaempferol and saline application groups (P>0.05).ConclusionsKaempferol application may be useful in decreasing alveolar bone resorption, attachment loss, and MMP-1 and -8 production in experimental periodontitis.

Effects of Zinc Supplementation on some Hematological and Biochemical Parameters in Liver Injured Rats

The present study was aimed to evaluate effect of zinc on some hematological and biochemical parameters in thioacitamide (TAA) induced liver cirrhotic rats. For this purpose 60 male Albino rats were divided into six groups (n=10). Group I, healthy control rats, Group II, received Zinc citrate (at a dose 5 mg/kg B.W, i.p , every other day for 8 weeks) , Group III, received Zinc sulfate (at a dose 10 mg/kg B.W, i.p , every other day for 8 weeks), Group IV, received thioacetamide (at a dose of 200mg/kg b.w, i.p, three time weekly for 8 weeks) ), Group V, received Zinc citrate and thioacitamide at the same time with the same dose of group II and group IV for 8 weeks, Group VI, received Zinc sulfate and thioacitamide at the same time with the same dose of group III and group IV for 8weeks. Some blood parameter (RBCs, HGB, HCT, Platelet, WBCs) were determined by using automatic blood counter apparatus also some biochemical parameters (GPT, GOT, ALP, GGT, Total protein and Albumin) were determined in serum. Results showed that in Zinc citrate and Zinc sulfate treated groups there is significant increase in RBCs, HGB and HCT values and significant decrease in WBCs while the thrombocyte values did not show any significant difference among the study groups as compared with control group at( P>.05).Also all blood parameters decrease in thioacitamide treated group as compared with control While zinc supplementation in combined treated groups decrease effect of thioacitamide in all blood parameters . furthermore, a significant increase in (GPT, GOT, ALP and GGT) with significant decrease of total protein and albumin in TAA treated group, However, treatment with zinc citrate and zinc sulfate and combined treatment show there is significant decrease in (GPT, GOT, ALP and GGT)with significant increase of total protein and albumin in groups treated with zinc citrate and zinc sulfate and also combined treated groups . Results indicate that zinc have a protective effect against the thioacetamide induced liver cirrhosis.

Studies on the biochemical and molecular effects of some natural herbs on experimental-induced breast cancer in Wistar rats

Background: Breast cancer is the most frequent malignant tumor in women, and its metastatic state represents the second cause of mortality. To treat cancer, we must prevent tumor cell proliferation and angiogenesis. The usage of herbal medication for treatment of some diseases as diabetes is increased nowadays for their wide safety margin. Aims and Objectives: Therefore, the current study was designed to examine the protective effect of ginger extract (GE) and cinnamon extract (CE) in controlling the mammary gland incidence in female Wistar rats. Materials and Methods: A total of 75 female virgin Wistar rats were allocated into 5 groups. Control group without any treatment; mammary gland group administered 7, 12-dimethyl-benz[a]anthracene at a dose of 20 mg/kg orally in corn oil. Rats in 3-5 served as tumor groups and received GE for group 3 (0.125% in water), CE for group 4 (100 mg/kg), and a mixture of GE plus CE for group 5. GE and CE were administered 2 weeks before cancer induction and continued for 4 months. Serum levels of breast cancer biomarker (CA125) and oxidative stress biomarkers were measured. Tissues of tumors were examined at molecular and histopathological levels. Results: There was a significant increase in the serum levels of CA125 in untreated mammary gland tumor group when compared with untreated healthy rats. There was a significant decrease in the serum levels of glutathione (GSH) peroxidase, catalase, superoxide dismutase, and GSH reductase in the untreated mammary gland tumor groups compared to control healthy rats. Administration of ginger and CE normalized the decrease in antioxidants levels. Induction of mammary gland tumor upregulated the genes associated with tumor incidences such as GST-P, CYP1A1, CYP1B1, and vascular endothelial growth factor-receptor 1 and administration of both herbal plants normalized these changes. Coadministration of GE and CE-induced additive inhibitory effect on genes upregulated due to tumor incidence. Bax expression was downregulated in tumor group and increased after GE and CE expression either alone or in combination. At histopathological levels, breast cancer group showed adenocarcinoma that is decreased with plants extracts administration when compared with breast cancer and control group. Conclusion: Results suggest that extracts of ginger and cinnamon have anticarcinogenic protective activities and support the hypothesis that these plants help in the prevention of mammary cancer by controlling the expression of carcinogenesis-associated genes.

Electroacupuncture Attenuates Visceral Pain and Reverses Upregulation of TRPV1 Expression in Adult Rats with Neonatal Maternal Deprivation

Irritable bowel syndrome (IBS) is characterized by chronic visceral hypersensitivity that companied by altered bowel movement. However, the treatment options are very limited. The aim of this study was to investigate effects of electroacupuncture (EA) on visceral hypersensitivity in a rat model of IBS and to explore the underlying mechanisms of EA effects. Visceral hypersensitivity was established by neonatal maternal deprivation (NMD) in male rats on postnatal days 2 - 15. Behavioral experiments were conducted at the age of 7 weeks. Treatment with EA at Zusanli (stomach-36, ST-36) significantly reduced abdominal withdrawal reflex (AWR) scores in NMD rats but not in age-matched healthy control rats. In addition, EA treatment hyperpolarized resting membrane potentials, increased the rheobase and reduced the numbers of action potentials evoked by 2 and 3 times rheobase current stimulation of dorsal root ganglion (DRG) neurons innervating the colon. NMD markedly enhanced expression of TRPV1 in colon related DRGs while EA treatment drastically suppressed the expression of TRPV1 in DRGs of NMD rats. These data suggest that EA treatment produced an analgesic effect, which might be mediated at least in a part by suppression of TRPV1 expression and by inhibition of neuronal excitability of primary sensory neurons in rats with chronic visceral pain.