ABSTRACT
OBJECTIVE@#To observe the effects of different suspension moxibustion methods on the syndrome characteristics and inflammatory factors of rats with rheumatoid arthritis (RA) of heat bi syndrome and to prove the concept of "moxibustion can be used for heat syndrome".@*METHODS@#Among seventy Wistar rats, 12 rats were randomly selected as a normal group, and the remaining rats were induced by collagen combined with wind, dampness, and heat environmental stimulation to establish the RA model of heat bi syndrome. Forty-eight rats with successful model establishment were further randomly divided into a model group and three moxibustion groups (mild moxibustion group, rotating moxibustion group and sparrow-pecking moxibustion group), with 12 rats in each group. The acupoints "Quchi" (LI 11), "Dazhui" (GV 14) and ashi point were used in all moxibustion groups, with mild moxibustion, rotating moxibustion, and sparrow-pecking moxibustion intervention given respectively, each acupoint was treated with moxibustion for 10 min a day, and 6 days were considered one course of treatment, with a total of three courses. After the intervention, the arthritis index (AI), the Evans blue (EB) extravasated volume in the soft tissue of the right hind paw, and the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-10 in the serum were measured by ELISA in each group. The volume of the bilateral hind paw was measured; the infrared thermal imaging was collected to analyze the temperature of the plantar area of the bilateral foot pads, and the reaction time of plantar heat pain was calculated before and after modeling, as well as after the 1st, 2nd and 3rd courses of interrention. The ankle dorsiflexion angle of the right hind foot was also measured before and after modeling, as well as after the intervention.@*RESULTS@#After modeling, compared with the normal group, the rats in the model group had more high-temperature areas in the bilateral hind limbs, abnormal AI score, abnormal bilateral hind paw volume, abnormal temperature of the plantar area of the bilateral foot pads, abnormal foot pain response time, abnormal right hind ankle dorsiflexion angle, abnormal right hind paw soft tissue EB extravasation, and abnormal serum TNF-α and IL-10 levels (P<0.01, P<0.05). After the intervention, compared with the model group, the rats in each moxibustion group had decreased or disappeared high-temperature areas in the bilateral hind limbs, EB extravasated volume in the soft tissue of the right hind paw was reduced (P<0.05), and the right ankle dorsiflexion angle was increased (P<0.05), serum level of TNF-α was reduced, and level of IL-10 increased (P<0.05); the AI scores in the mild moxibustion group and the sparrow-pecking moxibustion group was decreased (P<0.01, P<0.05). After the 1st, 2nd and 3rd courses of intervention, compared with the model group, the bilateral hind paw volume of rats in each moxibustion group was decreased (P<0.05, P<0.01), and plantar heat pain reaction time was increased (P<0.05). After the 2nd course and the 3rd course of intervention, the temperature of the right hind paw pad area was decreased in each moribustion group (P<0.05); after the 3rd courses of intervention, the temperature of the left hind paw pad area was decreased in the mild moxibustion group (P<0.05).@*CONCLUSION@#Suspension moxibustion could adjust the serum levels of TNF-α and IL-10 to improve the syndrome characteristics of RA rats of heat bi syndrome, such as joint redness, swelling, heat, pain and activity restriction. The effect of mild moxibustion is the most prominent. The findings could provide scientific basis for "moxibustion can be used for heat syndrome".
Subject(s)
Animals , Rats , Arthritis, Rheumatoid/therapy , Evans Blue , Hot Temperature , Interleukin-10/genetics , Moxibustion , Rats, Wistar , Tumor Necrosis Factor-alpha/geneticsABSTRACT
Objective: To investigate whether tanshinone ⅡA can protect the apoptosis of mice cochlear pericytes induced by high glucose and its specific protective mechanism, so as to provide experimental evidence for the prevention and treatment of diabetic hearing loss. Methods: C57BL/6J male mice were used to prepare type 2 diabetes model, which were divided into normal (NG) group, diabetic (DM) group, diabetic+tanshinone ⅡA (HG+tanshinone ⅡA) group and tanshinone ⅡA group. Each group had 10 animals. Primary cochlear pericytes were divided into NG group, HG group (high glucose 35 mmol/L), HG+tanshinone ⅡA (1, 3, 5 μmol/L) group, HG+Tanshinone ⅡA+LY294002 (PI3K/AKT pathway inhibitor) group, LY294002 group, tanshinone ⅡA group and DMSO group. Auditory brainstem response (ABR) was used to measure hearing threshold. Evans blue was used to detect the permeability of blood labyrinth barrier in each group. TBA methods were used to detect oxidative stress levels in various organs of mice. Morphological changes of stria vascularis were observed by hematoxylin-eosin staining (HE). Evans blue was used to detect the vascular labyrinth barrier permeability in cochlea. The expression of apoptosis protein in stria vascularis pericytes was observed by immunofluorescence. Pericytes apoptosis rate was observed by flow cytometry. DCFH-DA was combined with flow cytometry to detect intracellular ROS content, and Western blot was used to detect the expression of apoptotic proteins (Cleaved-caspase3, Bax), anti-apoptotic proteins (BCL-2) and pathway proteins (PI3K, p-PI3K, AKT, p-AKT). SPSS software was used for statistical analysis. Independent sample t test was performed, and P<0.05 was considered statistically significant. Results: Animal experiments: Tanshinone ⅡA decreased the hearing threshold of DM group [(35.0±3.5) dB SPL vs. (55.3±8.1) dB SPL] (t=4.899, P<0.01), decreased the oxidative stress level in cochlea (t=4.384, P<0.05), improved the structure disorder, atrophy of cochlea vascular lines, vacuole increased phenomenon. Tanshinone ⅡA alleviated the increased permeability of the blood labyrinth barrier [Evans blue leakage (6.84±0.27) AU vs. (8.59±0.85) AU] in the cochlea of DM mice (t=2.770, P<0.05), reversed the apoptotic protein: Caspase3 (t=4.956, P<0.01) and Bax (t=4.388, P<0.05) in cochlear vascularis. Cell experiments: Tanshinone ⅡA decreased intracellular ROS content in a concentration-dependent way (t=3.569, P<0.05; t=4.772, P<0.01; t=7.494, P<0.01); Tanshinone ⅡA decreased apoptosis rate and apoptotic protein, and increased the expression of anti-apoptotic protein, p-PI3K/PI3K and p-AKT/AKT in concentration-dependent manner (all P values<0.05); LY294002 reversed the protective effect of tanshinone ⅡA on pericytes apoptosis (all P values<0.05). Conclusion: Tanshinone ⅡA can inhibit the apoptosis of cochlear pericytes induced by high glucose by reducing oxidative stress level and activating PI3K/AKT signaling pathway under high glucose environment, thus playing a protective role in diabetic hearing loss.
Subject(s)
Animals , Male , Mice , Apoptosis , bcl-2-Associated X Protein , Diabetes Mellitus, Type 2 , Evans Blue , Glucose , Hearing Loss , Mice, Inbred C57BL , Pericytes/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Reactive Oxygen Species/metabolism , Signal TransductionABSTRACT
OBJECTIVE@#To determine whether Schisandrin B (Sch B) attenuates early brain injury (EBI) in rats with subarachnoid hemorrhage (SAH).@*METHODS@#Sprague-Dawley rats were divided into sham (sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B (100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan's blue extravasation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1 (Iba-1) and myeloperoxidase (MPO) in the rat brain, while the expressions of B-cell lymphoma 2 (Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in the rat brains were detected by Western blot.@*RESULTS@#Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan's blue content, and apoptotic cells number in the brain of rats (P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO (P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain (P<0.01), all of which were inhibited by Sch B (P<0.01). In addition, Sch B increased the Bcl-2 expression (P<0.01).@*CONCLUSION@#Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.
Subject(s)
Animals , Rats , Apoptosis , Brain/pathology , Brain Injuries/pathology , Caspase 3/metabolism , Cyclooctanes , Evans Blue , Inflammasomes/metabolism , Interleukin-18/metabolism , Lignans , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Polycyclic Compounds , Proto-Oncogene Proteins c-bcl-2/metabolism , Rats, Sprague-Dawley , Subarachnoid Hemorrhage/drug therapy , Water , bcl-2-Associated X Protein/metabolismABSTRACT
PURPOSE: Previous study has well documented the anti-apoptotic effects of miR-590 on oxidized low-density lipoprotein (ox-LDL)-treated endothelial cells (ECs). However, the mechanism underlying the anti-apoptotic effects of miR-590 in ox-LDL-treated ECs remains to be further addressed. MATERIALS AND METHODS: ApoE(−/−) mice fed with a high-fat diet (HFD) and human aortic endothelial cells (HAECs) treated with ox-LDL were used as in vivo and in vitro models of atherosclerosis. The expressions of miR-590 and toll-like receptor 4 (TLR4) were detected by quantitative real-time PCR and Western blot, respectively. Atherosclerotic lesion analysis was performed using Evans blue and hematoxylin-eosin staining. Cell proliferation was assessed by MTT assay. Apoptosis was examined using flow cytometry analysis and Western blot analysis of Cleaved poly (ADP-ribose) polymerase (PARP) and Cleaved Caspase-3 levels. The effect of miR-590 on TLR4/nuclear factor kappa B (NF-κB) pathway was evaluated by Western blot. Binding between miR-590 and TLR4 was confirmed by luciferase reporter assay and Western blot. RESULTS: miR-590 was downregulated in the aorta tissues from HFD-fed apoE(−/−) mice and ox-LDL-treated HAECs. miR-590 overexpression inhibited atherosclerotic lesion in HFD-induced apoE(−/−) mice and promoted proliferation and inhibited apoptosis of ox-LDL-treated HAECs. Additionally, TLR4 was identified as a direct target of miR-590 in ox-LDL-treated HAECs. Moreover, anti-miR-590 reversed TLR4 knockdown-mediated promotion of cell proliferation and suppression of apoptosis in ox-LDL-treated HAECs. miR-590 overexpression suppressed the TLR4/NF-κB pathway, and inhibition of the TLR4/NF-κB pathway promoted cell proliferation and impeded apoptosis in ox-LDL-treated HAECs. CONCLUSION: miR-590 promoted proliferation and blocked ox-LDL-induced apoptosis in HAECs through inhibition of the TLR4/NF-κB pathway.
Subject(s)
Animals , Humans , Mice , Aorta , Apoptosis , Atherosclerosis , Blotting, Western , Caspase 3 , Cell Proliferation , Diet, High-Fat , Endothelial Cells , Evans Blue , Flow Cytometry , In Vitro Techniques , Lipoproteins , Luciferases , Real-Time Polymerase Chain Reaction , Toll-Like Receptor 4ABSTRACT
In the twenty-first century, high contagious infectious diseases such as SARS (Severe Acute Respiratory Syndrome), MERS (Middle East Respiratory Syndrome), FMD (Foot-and-Mouth Disease) and AI (Avian Influenza) have become very prevalent, causing treat harm to humans and animals in aspect of public health, and economical issues. The critical problem is that newly-reported infectious diseases that humans firstly experience are expected to continue to emerge, and these diseases will be spreading out rapidly. Therefore, rapid and safe supplies of effective vaccines are most pivotal to prevent the rapid prevalent of new infection, but international standards or assessing protocol the safety of urgent vaccines are not established well. In our previous study, since we established a module to assess the brain safety of urgent vaccines, therefore, it is necessary to verify that this established module for assessing brain safety could work effectively in commercially available two vaccines (one killed- and on live-vaccines). We compared the results of Evans blue (EB) assay and qPCR analysis by injection of two kinds of vaccines, PBS and Lipopolysaccharide (LPS) under the condition of the module previously reported. We confirmed that the brain safety test module for urgent vaccine we established is very reproducible. Therefore, it is believed that this vaccine safety testing method can be used to validate brain safety when prompt supply of a newly developed vaccines is needed.
Subject(s)
Animals , Humans , Brain , Communicable Diseases , Coronavirus Infections , Equipment and Supplies , Evans Blue , Methods , Public Health , Severe Acute Respiratory Syndrome , VaccinesABSTRACT
PURPOSE: Patients treated with propranolol, a nonselective β-adrenoceptor antagonist, develop severe anaphylaxis, but the mechanism remains unknown. We determined effects of β₁- and β₂-adrenoceptor antagonists on the anaphylaxis-induced increase in vascular permeability in mice. METHODS: In anesthetized ovalbumin-sensitized C57BL mice, mean arterial blood pressure (MBP) was measured, and Evans blue dye extravasation and hematocrit (Hct) were assessed at 20 minutes after antigen injection. The following pretreatment groups (n=7/group) were studied: (1) sensitized control (non-pretreatment), (2) propranolol, (3) the selective β₂-adrenoceptor antagonist ICI 118,551, (4) the selective β₁-adrenoceptor antagonist atenolol, (5) adrenalectomy, (6) the selective β₂-adrenoceptor agonist terbutaline, and (7) non-sensitized groups. RESULTS: The antigen injection decreased MBP, and increased Hct and vascular permeability in the kidney, lung, mesentery, and intestine, but not in the liver or spleen. Pretreatment with ICI 118,551, propranolol and adrenalectomy, but not atenolol, reduced the survival rate and augmented the increases in Hct and vascular permeability in the kidney, intestine, and lung as compared with the sensitized control group. Pretreatment with terbutaline abolished the antigen-induced alterations. Plasma epinephrine levels were increased significantly in the sensitize control mice. CONCLUSIONS: Blockade of β₂-adrenoceptor can deteriorate systemic anaphylaxis by augmenting hyperpermeability-induced increase in plasma extravasation by inhibiting beneficial effects of epinephrine released from the adrenal glands in anesthetized mice.
Subject(s)
Animals , Humans , Mice , Adrenal Glands , Adrenalectomy , Anaphylaxis , Arterial Pressure , Atenolol , Capillary Permeability , Epinephrine , Evans Blue , Hematocrit , Intestines , Kidney , Liver , Lung , Mesentery , Mice, Inbred C57BL , Plasma , Propranolol , Spleen , Survival Rate , TerbutalineABSTRACT
PURPOSE: Microvascular endothelial integrity is important for maintaining the blood-brain barrier (BBB). However, subarachnoid hemorrhage (SAH) disrupts this integrity, making the BBB dysfunctional—an important pathophysiological change after SAH. Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) regulate microvascular permeability by balancing each other’s expression. METHODS: This study investigated the dynamics of Ang-1 and Ang-2 expression after SAH and the protective effect of Ang-1 on BBB functioning using an endovascular puncture model of rat SAH. The Ang-1 and Ang-2 expression in brain tissue was determined by immunohistochemistry. In addition, Western blotting was used to estimate Ang-1 and Ang-2 concentration and to compare them at 6–72 hours post-SAH cortex and hippocampus. Evans blue viability assay was used to evaluate BBB permeability, and neurological testing was implemented to evaluate neurological impairment during SAH. RESULTS: It was found that following SAH, Ang-1 expression decreases and Ang-2 expression increases in the cortex, hippocampus, and microvessels. The Ang-1/Ang-2 ratio decreased as quickly as 6 hours after SAH and reached its lowest 1 day after SAH. Finally, it was found that exogenous Ang-1 reduces SAH-associated BBB leakage and improves neurological function in post-SAH rats. CONCLUSIONS: Our findings suggest that the equilibrium between Ang-1 and Ang-2 is broken in a period shortly after SAH, and the treatment of exogenous Ang-1 injection alleviates neurological dysfunctions through decreasing BBB destruction.
Subject(s)
Animals , Rats , Angiopoietin-1 , Angiopoietin-2 , Blood-Brain Barrier , Blotting, Western , Brain , Brain Injuries , Capillary Permeability , Evans Blue , Hippocampus , Immunohistochemistry , Microvessels , Permeability , Punctures , Subarachnoid HemorrhageABSTRACT
Dyes are the most difficult constituents to remove by conventional biological wastewater treatment. Colored wastewater is mainly eliminated by physical and chemical procedures, which are very expensive and have drawbacks. Therefore, the advantage of using biological processes, such as the biotransformation of dyes, is that they may lead to complete mineralization or formation of less toxic products. To prove the possibility of using fungal processes for decolorization and other applications, the analysis of the toxicity of the processes' products is required. The decolorization of the mixture of two dyes from different classes - triphenylmethane brilliant green and azo Evans blue (GB - total concentration 0.08 g/L, proportion 1:1 w/w) - by Pleurotus ostreatus (BWPH and MB), Gloeophyllum odoratum (DCa), RWP17 (Polyporus picipes) and Fusarium oxysporum (G1) was studied. Zootoxicity (Daphnia magna) and phytotoxicity (Lemna minor) changes were estimated at the end of the experiment. The mixture of dyes was significantly removed by all the strains that were tested with 96 h of experimental time. However, differences among strains from the same species (P. ostreatus) were noted. Shaking improved the efficacy and rate of the dye removal. In static samples, the removal of the mixture reached more than 51.9% and in shaken samples, more than 79.2%. Tests using the dead biomass of the fungi only adsorbed up to 37% of the dye mixture (strain BWPH), which suggests that the process with the living biomass involves the biotransformation of the dyes. The best results were reached for the MB strain, which removed 90% of the tested mixture under shaking conditions. Regardless of the efficacy of the dye removal, toxicity decreased from class V to class III in tests with D. magna. Tests with L. minor control samples were classified as class IV, and samples with certain strains were non-toxic. The highest phytotoxicity decrease was noted in shaken samples where the elimination of dye mixture was the best.
Subject(s)
Animals , Basidiomycota/growth & development , Basidiomycota/metabolism , Evans Blue/metabolism , Fusarium/growth & development , Fusarium/metabolism , Rosaniline Dyes/metabolism , Wastewater/microbiology , Araceae/drug effects , Araceae/physiology , Biotransformation , Cell Survival/drug effects , Daphnia/drug effects , Daphnia/physiology , Evans Blue/toxicity , Rosaniline Dyes/toxicity , Water Purification/methodsABSTRACT
OBJECTIVE: The purpose of this study was to correlate permeability parameters measured with dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) using a clinical 3-tesla scanner with extravasation of Evans blue in a rat model with transient cerebral ischemia. MATERIALS AND METHODS: Sprague-Dawley rats (n = 13) with transient middle cerebral artery occlusion were imaged using a 3-tesla MRI with an 8-channel wrist coil. DCE-MRI was performed 12 hours, 18 hours, and 36 hours after reperfusion. Permeability parameters (K(trans), v(e), and v(p)) from DCE-MRI were calculated. Evans blue was injected after DCE-MRI and extravasation of Evans blue was correlated as a reference with the integrity of the blood-brain barrier. Correlation analysis was performed between permeability parameters and the extravasation of Evans blue. RESULTS: All permeability parameters (K(trans), v(e), and v(p)) showed a linear correlation with extravasation of Evans blue. Among them, K(trans) showed highest values of both the correlation coefficient and the coefficient of determination (0.687 and 0.473 respectively, p < 0.001). CONCLUSION: Permeability parameters obtained by DCE-MRI at 3-T are well-correlated with Evans blue extravasation, and K(trans) shows the strongest correlation among the tested parameters.
Subject(s)
Animals , Male , Rats , Blood-Brain Barrier/pathology , Capillary Permeability , Contrast Media , Disease Models, Animal , Evans Blue/analysis , Ischemic Attack, Transient/diagnosis , Magnetic Resonance Imaging/instrumentation , Rats, Sprague-Dawley , Stroke/diagnosisABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective effects of Jiedu Tongluo injection on cerebral edema induced by focal lesion of cerebral ischemia/reperfusion, the hydrous content of brain and the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and MMP-9 in rats.</p><p><b>METHOD</b>The model of brain middle cerebral artery ischemia/reperfusion was established by the thread approach. After 24 hours of reperfusion, cerebral edema formation was determined by the hydrous content of brain. The permeability of blood brain barrier was evaluated based on the leakage of Evans blue. Enzyme-linked immunoadsordent assay (ELISA)was used to examine the expression of ICAM-1, VCAM-1, E-selectin. The expression of MMP-9 was measured by immunohistochemistry.</p><p><b>RESULT</b>JDTL, in the dose of 2 mL x kg(-1) and 4 mL x kg(-1), relieved cerebral edema (P < 0.05, P < 0.01), reduced the expressions of ICAM-1, VCAM-land E-selectin and decreased MMP-9 activity (P < 0. 05, P < 0.01) in model rats.</p><p><b>CONCLUSION</b>Jiedu Tongluo injection has a protective effect on rat brain from cerebral edema induced by the injury of focal cerebral ischemia/reperfusion. The mechanism is related to that Jiedu Tongluo injection can reduce the expressions of ICAM-1, VCAM-1 and E-selectin and inhibit of MMP-9 activation in rat brain.</p>
Subject(s)
Animals , Male , Rats , Blood-Brain Barrier , Metabolism , Brain Edema , Metabolism , Brain Ischemia , Drugs, Chinese Herbal , Pharmacology , E-Selectin , Metabolism , Evans Blue , Metabolism , Gene Expression Regulation, Enzymologic , Injections , Intercellular Adhesion Molecule-1 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Permeability , Rats, Sprague-Dawley , Reperfusion Injury , Vascular Cell Adhesion Molecule-1 , MetabolismABSTRACT
BACKGROUND/OBJECTIVE: Myocardial cell death due to occlusion of the coronary arteries leads to myocardial infarction, a subset of coronary heart disease (CHD). Dietary fiber is known to be associated with a reduced risk of CHD, the underlying mechanisms of which were suggested to delay the onset of occlusion by ameliorating risk factors. In this study, we tested a hypothesis that a beneficial role of dietary fiber could arise from protection of myocardial cells against ischemic injury, manifested after occlusion of the arteries. MATERIALS/METHODS: Three days after rats were fed apple pectin (AP) (with 10, 40, 100, and 400 mg/kg/day), myocardial ischemic injury was induced by 30 min-ligation of the left anterior descending coronary artery, followed by 3 hr-reperfusion. The area at risk and infarct area were evaluated using Evans blue dye and 2,3,5-triphenyltetrazolium chloride (TTC) staining, respectively. DNA nicks reflecting the extent of myocardial apoptosis were assessed by TUNEL assay. Levels of cleaved caspase-3, Bcl-2, and Bax were assessed by immunohistochemistry. RESULTS: Supplementation of AP (with 100 and 400 mg/kg/day) resulted in significantly attenuated infarct size (IS) (ratio of infarct area to area at risk) by 21.9 and 22.4%, respectively, in the AP-treated group, compared with that in the control group. This attenuation in IS showed correlation with improvement in biomarkers involved in the apoptotic cascades: reduction of apoptotic cells, inhibition of conversion of procaspase-3 to caspase-3, and increase of Bcl-2/Bax ratio, a determinant of cell fate. CONCLUSIONS: The findings indicate that supplementation of AP results in amelioration of myocardial infarction by inhibition of apoptosis. Thus, the current study suggests that intake of dietary fiber reduces the risk of CHD, not only by blocking steps leading to occlusion, but also by protecting against ischemic injury caused by occlusion of the arteries.
Subject(s)
Animals , Rats , Apoptosis , Arteries , Biomarkers , Caspase 3 , Cell Death , Coronary Disease , Coronary Vessels , Dietary Fiber , DNA Breaks, Single-Stranded , Evans Blue , Immunohistochemistry , In Situ Nick-End Labeling , Ischemia , Models, Animal , Myocardial Infarction , Risk FactorsABSTRACT
BACKGROUND/OBJECTIVE: Myocardial cell death due to occlusion of the coronary arteries leads to myocardial infarction, a subset of coronary heart disease (CHD). Dietary fiber is known to be associated with a reduced risk of CHD, the underlying mechanisms of which were suggested to delay the onset of occlusion by ameliorating risk factors. In this study, we tested a hypothesis that a beneficial role of dietary fiber could arise from protection of myocardial cells against ischemic injury, manifested after occlusion of the arteries. MATERIALS/METHODS: Three days after rats were fed apple pectin (AP) (with 10, 40, 100, and 400 mg/kg/day), myocardial ischemic injury was induced by 30 min-ligation of the left anterior descending coronary artery, followed by 3 hr-reperfusion. The area at risk and infarct area were evaluated using Evans blue dye and 2,3,5-triphenyltetrazolium chloride (TTC) staining, respectively. DNA nicks reflecting the extent of myocardial apoptosis were assessed by TUNEL assay. Levels of cleaved caspase-3, Bcl-2, and Bax were assessed by immunohistochemistry. RESULTS: Supplementation of AP (with 100 and 400 mg/kg/day) resulted in significantly attenuated infarct size (IS) (ratio of infarct area to area at risk) by 21.9 and 22.4%, respectively, in the AP-treated group, compared with that in the control group. This attenuation in IS showed correlation with improvement in biomarkers involved in the apoptotic cascades: reduction of apoptotic cells, inhibition of conversion of procaspase-3 to caspase-3, and increase of Bcl-2/Bax ratio, a determinant of cell fate. CONCLUSIONS: The findings indicate that supplementation of AP results in amelioration of myocardial infarction by inhibition of apoptosis. Thus, the current study suggests that intake of dietary fiber reduces the risk of CHD, not only by blocking steps leading to occlusion, but also by protecting against ischemic injury caused by occlusion of the arteries.
Subject(s)
Animals , Rats , Apoptosis , Arteries , Biomarkers , Caspase 3 , Cell Death , Coronary Disease , Coronary Vessels , Dietary Fiber , DNA Breaks, Single-Stranded , Evans Blue , Immunohistochemistry , In Situ Nick-End Labeling , Ischemia , Models, Animal , Myocardial Infarction , Risk FactorsABSTRACT
The structure of coxsackievirus and adenovirus receptor's CAR is similar to adhesion molecules. In the adult heart, the majority of CAR localizes at the intercalated disc. Germ line CAR deletion induces embryonic lethality at E11.5 with evidence of a cardiac abnormality. The CAR role as a viral receptor is well known; however, its precise function in the heart for enterovirus infection is not clear. To understand the role of CAR in the cardiac myocyte, we generated cardiac-specific CAR knockout mice using a CAR floxed allele and alpha-MHC-Mer CRE Mer mice. Western blot analysis and immunofluorescent stain of ventricles at 6 weeks after 2 weeks tamoxifen administration, CAR expression was significantly decreased in CAR(f/f) MCM mice but not in CAR(f/f) mice heart. Enterovirus was intraperitoneally infected into CAR(f/f) MCM and CAR(f/f) mice (n=10 each). CAR disruption was dramatically reduced virus infection and replication in the heart but not different in liver, spleen, and pancreas. Cardiac myocyte damage was significantly reduced in the CAR(f/f) MCM mutant mice by evans blue dye stain. In addition, the CAR(f/f) MCM mutant mice heart inflammation and fibrosis were decreased in H&E and trichrome stain compare to CAR(f/f) control mice. CAR expression was required for normal ventricular function, but it is the cause of enterovirus infection. In the adult mice heart, CAR deletion was significantly reduced viral infection, proliferation, and myocarditis. These results suggested that CAR deletion could be useful therapeutic strategy to prevent viral myocarditis.
Subject(s)
Adult , Animals , Humans , Mice , Adenoviridae , Alleles , Azo Compounds , Blotting, Western , Enterovirus , Enterovirus Infections , Eosine Yellowish-(YS) , Evans Blue , Fibrosis , Germ Cells , Heart , Inflammation , Liver , Methyl Green , Mice, Knockout , Myocarditis , Myocytes, Cardiac , Pancreas , Receptors, Virus , Spleen , Tamoxifen , Ventricular Function , VirusesABSTRACT
OBJECTIVE@#To observe changes in expression of tumor necrosis factor (TNF)-alpha and permeability of blood brain barrier after salidroside pretreatment in rats with injury induced by focal cerebralischemia-reperfusion.@*METHODS@#Forty-five male SD rats were randomly divided into three groups (n=15): control group, ischemia-reperfusion (IR) model group, and salidroside pretreatment group. Before the IR model establishment, the rats in the salidroside pretreatment group were intraperitoneally administered with salidroside at a dose of 24 mg/(kg·d) for 7 d. After 30 min post the last administration, the IR model was induced by occlusion of middle cerebral artery with a filament. After 24 h post the operation, the water content and Evens blue content in the ischemia cerebral hemisphere were determined, and the level of TNF-alpha mRNA was detected by the semi-quantitative RT-PCR.@*RESULTS@#Compared with the IR model group, the salidroside pretreatment group had significantly lower (P<0.05) water content and Evens blue content in the ischemia cerebral hemisphere and also had significantly lower (P<0.05) level of TNF-alpha in the ischemic cerebral cortex tissue.@*CONCLUSIONS@#The salidroside pretreatment alleviated the focal cerebralischemia-reperfusion injury in the rat model, possibly by decreasing the permeability of blood brain barrier, attenuating brain edema and reducing TNF-alpha expression.
Subject(s)
Animals , Male , Rats , Blood-Brain Barrier , Metabolism , Brain Ischemia , Drug Therapy , Metabolism , Cerebral Cortex , Chemistry , Metabolism , Disease Models, Animal , Evans Blue , Chemistry , Gene Expression , Glucosides , Pharmacology , Phenols , Pharmacology , Protective Agents , Pharmacology , Random Allocation , Rats, Sprague-Dawley , Reperfusion Injury , Metabolism , Tumor Necrosis Factor-alpha , Genetics , MetabolismABSTRACT
OBJECTIVE: To reveal the expression and possible roles of aquaporin 9 (AQP9) in rat brain, after severe traumatic brain injury (TBI). METHODS: Brain water content (BWC), tetrazolium chloride staining, Evans blue staining, immunohistochemistry (IHC), immunofluorescence (IF), western blot, and real-time polymerase chain reaction were used. RESULTS: The BWC reached the first and second (highest) peaks at 6 and 72 hours, and the blood brain barrier (BBB) was severely destroyed at six hours after the TBI. The worst brain ischemia occurred at 72 hours after TBI. Widespread AQP9-positive astrocytes and neurons in the hypothalamus were detected by means of IHC and IF after TBI. The abundance of AQP9 and its mRNA increased after TBI and reached two peaks at 6 and 72 hours, respectively, after TBI. CONCLUSIONS: Increased AQP9 might contribute to clearance of excess water and lactate in the early stage of TBI. Widespread AQP9-positive astrocytes might help lactate move into neurons and result in cellular brain edema in the later stage of TBI. AQP9-positive neurons suggest that AQP9 plays a role in energy balance after TBI.
OBJETIVO: Revelar a expressão e os possíveis papéis da aquaporina 9 (AQP9) no cérebro de ratos após lesão cerebral traumática (LCT) grave. MÉTODOS: Foram utilizados: determinação do conteúdo cerebral de água, corante cloreto de tetrazólio, corante azul de Evans, imunoistoquímica (IHQ), imunofluorescência (IF), western blot e PCR em tempo real. RESULTADOS: O conteúdo cerebral de água alcançou o primeiro e o segundo (o mais alto) picos após 6 e 72 horas. A função da barreira hematoencefálica se mostrou muito prejudicada após 6 horas da LCT. A pior isquemia cerebral ocorreu após 72 horas da LCT. Astrócitos AQP9 positivos e neurônios no hipotálamo foram detectados difusamente pela IHQ e IF após LCT. A abundância de AQP9 e de sua mRNA aumentou após LCT e alcançou dois picos após 6 e 72 horas, respectivamente, da LCT. CONCLUSÕES: AQP9 aumentada pode contribuir para a eliminação de água e lactato em excesso na fase precoce da LCT. Astrócitos difusamente localizados AQP9 positivos podem ajudar a entrada do lactato nos neurônios, promovendo edema cerebral celular na fase tardia da LCT. Neurônios AQP9 positivos sugerem que AQP9 tem um papel no equilíbrio energético após LCT.
Subject(s)
Animals , Male , Rats , Aquaporins/metabolism , Brain Injuries/metabolism , Blotting, Western , Evans Blue , Fluorescent Antibody Technique , Immunohistochemistry , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Severity of Illness Index , Staining and Labeling , Tetrazolium SaltsABSTRACT
PURPOSE: The present study investigated the effects of triamcinolone acetonide (TA) on retinal vessel-related factors and retinal vascular leakage in the retina of oxygen-induced retinopathy (OIR) rats. METHODS: Sprague-Dawley rats used for OIR were exposed to hyperoxia from postnatal day 2 to day 14, then returned to normoxia from day 15 to day 30 and compared with control rats. On postnatal day 16, 2 microl of TA (4 mg/kg) was injected into the vitreous of the right eye in control and OIR rats. The Evans blue method was used for evaluating vascular leakage and RT-PCR was performed for confirmation of mRNA expression. RESULTS: In OIR rats exposed to hyperoxia, retinal vascular permeability increased when returned to normoxia. After intravitreal injection of TA, vascular permeability was decreased in OIR rats, but no changes were found in control rats. In OIR rats, mRNAs of HIF-1alpha, VEGF, SDF-1 and ICAM-1 were more expressed and down-regulated after intravitreal injection of TA. Occludin mRNA level was decreased in the hypoxic condition and up-regulated after TA treatment. CONCLUSIONS: The present study suggests the ability of TA to inhibit retinal vascular leakage in OIR rats and a possibility that TA controls expression of the HIF-1, VEGF, SDF-1, ICAM-1 and Occludin, which may protect retinal vascular destruction from hypoxic conditions; further studies are necessary to confirm changes in protein levels.
Subject(s)
Animals , Rats , Capillary Permeability , Evans Blue , Eye , Hyperoxia , Intercellular Adhesion Molecule-1 , Intravitreal Injections , Occludin , Rats, Sprague-Dawley , Retina , Retinaldehyde , RNA, Messenger , Triamcinolone , Triamcinolone Acetonide , Vascular Endothelial Growth Factor AABSTRACT
PURPOSE: The present study investigated the effects of triamcinolone acetonide (TA) on retinal vessel-related factors and retinal vascular leakage in the retina of oxygen-induced retinopathy (OIR) rats. METHODS: Sprague-Dawley rats used for OIR were exposed to hyperoxia from postnatal day 2 to day 14, then returned to normoxia from day 15 to day 30 and compared with control rats. On postnatal day 16, 2 microl of TA (4 mg/kg) was injected into the vitreous of the right eye in control and OIR rats. The Evans blue method was used for evaluating vascular leakage and RT-PCR was performed for confirmation of mRNA expression. RESULTS: In OIR rats exposed to hyperoxia, retinal vascular permeability increased when returned to normoxia. After intravitreal injection of TA, vascular permeability was decreased in OIR rats, but no changes were found in control rats. In OIR rats, mRNAs of HIF-1alpha, VEGF, SDF-1 and ICAM-1 were more expressed and down-regulated after intravitreal injection of TA. Occludin mRNA level was decreased in the hypoxic condition and up-regulated after TA treatment. CONCLUSIONS: The present study suggests the ability of TA to inhibit retinal vascular leakage in OIR rats and a possibility that TA controls expression of the HIF-1, VEGF, SDF-1, ICAM-1 and Occludin, which may protect retinal vascular destruction from hypoxic conditions; further studies are necessary to confirm changes in protein levels.
Subject(s)
Animals , Rats , Capillary Permeability , Evans Blue , Eye , Hyperoxia , Intercellular Adhesion Molecule-1 , Intravitreal Injections , Occludin , Rats, Sprague-Dawley , Retina , Retinaldehyde , RNA, Messenger , Triamcinolone , Triamcinolone Acetonide , Vascular Endothelial Growth Factor ASubject(s)
Animals , Male , Rats , Muscle, Skeletal/injuries , Diabetes Complications , Immunity , Cytokines/blood , Rats, Wistar , Evans BlueABSTRACT
Busca-se em pesquisas e estudos avaliar a capacidade de adaptação e selamento entre a conexão implante/intermediário de diferentes sistemas de implantes odontológicos. Observou-se recentemente que implantes com abutments retidos com parafusos, diversos fenômenos como afrouxamento e fratura do parafuso, rotação e fratura do abutment com penetração bacteriana nas câmaras internas dos implantes, acontece como conseqüência da desadaptação interface implante/abutment. É descrito ao nível desta região um pequeno espaço microgap, fator relevante para remodelamento da crista óssea e longevidade da saúde dos tecidos moles periimplantares. O propósito do estudo foi investigar o extravasamento da solução do corante azul de Evan em três tipos de implantes e seus respectivos intermediários, durante um período de seis (6) dias, a cada vinte e quatro (24) horas, com intervalo em cento e vinte (120) horas, através da agitação proporcionada por uma mesa agitadora. Para tal, foram utilizados trinta (30) implantes, dez (10) de cada tipo, com seus respectivos intermediários protéticos, minipilares, sendo o Grupo Um (1) de implantes Hexágono Externo (HE), Grupo dois (2) de Hexágono Interno (HI) e Grupo três (3) de Cone Morse (CM). No interior de cada implante foi pipetado volume ou quantidade proporcional ao seu espaço interno uma solução de corante azul de Evan. Após a colocação do corante no interior dos implantes, os abutments ou intermediários foram acoplados e aparafusados com torque de vinte (20) Ncm, através do torquímetro de Gauge (Tohnichi), e estes depositados individualmente em micro tubos de cor âmbar na condição de intermediários voltados para baixo. Segui/se imediatamente a colocação de (1)ml de água deionizada. A seguir os tubos foram fechados hermeticamente e posicionados numa mesa suporte para microtubos e foram armazernados por 24 horas, sem agitação. Posteriormente foram agitados por 10 minutos com movimentos uniformes em mesa agitadora e a partir deste momento...
Research and studies seek to evaluate the capacity of adaptation and sealing between the implant-intermediate connections of different dental implant systems. It has recently been observed that in implants with screw-retained abutments, various phenomena, such as screw4 loosening and fracture, rotation and fracture of the abutment with bacterial penetration into the internal chambers of the implants have occurred as a result of maladaptatation at the implant-abutment interface. At the level of this region, a small space known as a microgap is described, and is a relevant factor in remodeling of the crestal bone and peri-implant soft tissue health in the long term. The purpose of this study was to investigate the extravasation of Evans blue dye solution in three types of implants and their respective intermediates during a period of six (6) days, every twenty-four (24) hours, with an interval in one hundred and wenty (120) hours, by means of agitation provided by an agitating table. To do this, thirty (30) implants were used, ten (10) of each type, with their respective prosthetic intermediates and mini-abutments, divided into groups as follows: Group One (1) External Hexagon implants (EH), Group Two (2) Internal Hexagon (IH) and Group three (3) Morse Cone (MC). Into the interior of each implant, Evans blue dye solution was inserted with a pipette in a volume or quantity proportional to its internal space. After the dye was put into the implants, the abutments or intermediates were coupled and the screws tightened to a torque of twenty (20) Ncm, with a Gauge torque meter (Tohnichi), and they were individually deposited in amber-colored microtubes positioned so that the intermediates faced downwards. This was immediately followed by the placement of (1)ml of deionized water. Next, the tubes were hermetically closed and placed on a table with a microtube holder and stored for 24 hours, without agitation. Afterwards they were agitated for 10...
Subject(s)
Coloring Agents/chemistry , Evans Blue/chemistry , Dental Pulp Cavity , Dental Prosthesis, Implant-Supported/methods , Analysis of Variance , Materials Testing , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells (ECs) with the protein expression of zonula occludens-1 (ZO-1).</p><p><b>METHODS</b>Sprague Dawley rats were randomly divided into a model group of severe acute pancreatitis (SAP) and a sham-operated (SO) group. The SAP rats were further divided into two subgroups on the basis of tongue-coating status: a thick greasy tongue fur group (SAP-TGF) and a normal tongue fur group (SAP-NF). Six rats were chosen randomly from every group mentioned above for an Evans blue assay 5 days after model establishment. For the histomorphology analysis, the expressions of ZO-1 protein and mRNA were studied by hematoxylin-eosin (H&E) staining, transmission electron microscope, Western blot, and Q-PCR using blood and tongue tissues, which were collected from 8 rats randomly chosen from each group.</p><p><b>RESULTS</b>The papillae density of the rat tongue surface and the caryocinesis frequency of the basal layer were significantly increased in the SAP-TGF group compared with the SO group (P<0.05). Evans blue levels in the tongue tissue of the SAP-TGF group were significantly higher than that of the SO and SAP-NF groups (P<0.05). Vascular ECs were wider and obviously fissured in the SAP-TGF group under transmission electron microscope observation. The protein and mRNA expression of ZO-1 in the SAP-TGF group were lower than those in the SAP-NF (P<0.05).</p><p><b>CONCLUSIONS</b>Reproductive activity enhancement of glossal epithelial cells was one of the main characteristics of thick greasy tongue fur formation. An increase in vasopermeability was closely associated with thick greasy tongue fur formation. Tight junction structural variation of vascular ECs might play an important role in the pathological and physiological process of thick greasy tongue fur formation.</p>