Effects of Shenlian Extracts () on Atherosclerosis by Inhibition of the Inflammatory Response

ObjectiveInflammation plays a critical role in atherosclerosis, and this inflammatory reaction is being intensively studied. Shenlian Extracts (), an active ingredient of Chinese medicinal herbs, is believed to have multiple therapeutic and preventive effects against human vascular diseases, including atherosclerosis. Our work investigated whether Shenlian Extracts serves as an anti-inflammatory agent during atherogenesis.MethodsWe established a model of atherosclerosis in rabbits using balloon angioplasty and a high cholesterol diet. The effects of Shenlian Extracts on vessel structure and inflammation were assessed by hematoxylin-eosin staining of the femoral artery, measurement of inflammation-related factors in serum or vascular tissue, and radioimmunoassay. Enzyme linked immunosorbent assays (ELISA), flow cytometry and western blots were also performed.ResultsWe show that oral pre-treatment with Shenlian Extracts suppressed the pathological changes associated with atherosclerosis and that graded doses of Shenlian Extracts reduced total serum levels of cholesterol (90, 180 and 360 mg/kg), triglyceride (180 and 360 mg/kg), and LDL-c (90, 180 mg/kg). Various doses of Shenlian Extracts reduced serum content of TNF-a (180 and 360 mg/kg), CRP (90, 180 and 360 mg/kg) and IL-8 (360 mg/kg) (P<0.05), but led to no significant changes in IL-1β levels. Treatment with Shenlian Extracts also significantly reduced VCAM-1 levels (90 and 360 mg/kg) and IGF-1 levels (90 and 180 mg/kg) in vascular tissue but had no significant effect on ICAM-1 and MCP-1 levels. Finally, Shenlian Extracts significantly reduced the abnormal expression of CD18 in monocytes in a dose-dependent manner.ConclusionThese results suggest that Shenlian Extracts may play a direct role in preventing and treating atherogenesis by inhibiting the inflammatory reaction, providing insights into the possible mechanism underlying the anti-atherosclerotic actions of Shenlian Extracts

Analysis of the Histopathology, TNF-α of Microglia Cells Expression, NRG-1/erbB Oligodendrocyte, and Ki67/Apoptosis of Dentate Gyrus Rattus novergicus Brain After Acute Traumatic Brain Injury

Head trauma or traumatic brain injury (TBI) gives most serious impact on the central nervous system. Several experimental models have been established to mimic different pathogenesis characteristics of TBI. The purpose of this study was to determine whether there is evidence of hystopathological lesions in the brain tissue after Marmorou TBI models. This study uses Rattus norvegicus Sprague Dawley strain. Macroscopic and microscopic observations on the brain tissue were done. Macroscopic lesions were observed in the brain. Microscopic observation was performed with Haematoxylin-Eosin (HE) staining and immunohistochemistry on the distribution of microglia cells and pyramidal cells in the cortex. Meanwhile, the distribution of NRG-1/ErbB, proliferation, and apoptosis were observed in the hippocampus. The results of macroscopic observation showed that there were wounds caused by falling loads and vasodilatation. On microscopic observation, the TBI group showed an increase in neutrophils distribution and distribution of activated microglia to produce TNF-α, and decrease in the number of cortical pyramidal cells significantly. The distribution of NRG-1 tended to decrease after exposure of TBI and had no effect on its receptor, erbB. Exposure of TBI appears to lower the activity of neuronal cells proliferation in dentate gyrus (DG) area and significantly increase the number of apoptotic cells. Marmarou model is a physiological model of TBI that spontaneously occurs following a trauma to the head, for example trauma due to an accident. This data can be used as a preliminary data of inflammation and tissue regeneration of disrupted adult brain. Therefore, this research could be used as the basis in the studies of therapeutic agents in the process of neurogenesis of brain cells.Keywords: traumatic brain injury, ERG-1/ErbB, dentate gyrus, Ki67, TNF-a, microgli

Role of Caspase-8 on Human Retinal Microvasculature Endothelial Cells (HRMEC) Migration in Hyperglycemia

Pathological angiogenesis is the hallmark of proliferative diabetic retinopathy that results from the advanced stage of Diabetic Retinopathy (DR). In response to prolonged tissue injury, retinal microvascular endothelial cells form new blood vessels to ensure the supply of oxygen and nutrients to the eye; however, progressive fibrovascular proliferation can eventually lead to retinal detachment and blindness. Caspase-8, the responsible protease for apoptosis has also been shown to modulate pathological and developmental angiogenesis but whether it exerts this role in a hyperglycemic environment has not yet been explored. Here we demonstrate that Caspase-8 decreased cell viability and was upregulated in hyperglycemic HRMEC. In addition, Caspase-8 K.D. resulted in inhibition of cell migration. Taken together, our data suggest that Caspase-8 plays a role that is involved in cell migration in HRMEC

Effects of Cyclic Heat Stress on the Acute Inflammatory Response in Broilers

Heat stress (HS) is a growing concern in broiler production due to increasing environmental temperatures. Little is known of the overall effect of HS on innate immunity. Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, is commonly used to study the inflammatory response. In avian species, the local tissue and systemic inflammatory activities in response to LPS may be determined concurrently, over time, in an individual, using the growing feather (GF) pulp dermal test along with blood measurements (dual-window approach). To examine the effect of cyclic HS on the local and systemic acute inflammatory responses, Cobb 500 male broiler chicks were reared under thermoneutral (TN) or cyclic HS conditions. Eight environmental chambers were used, four TN and four HS, with each chamber split into two pens. Beginning at 4 d of age, cyclic HS birds were subjected to 35°C from 8 am to 10 pm and TN temperatures from 10 pm to 8 am. At 37 d of age, four groups of broilers were formed: TN-LPS (n = 8 broilers), TN-PBS (phosphate-buffered saline (vehicle) control; n = 4), HS-LPS (n = 8) and HS-PBS (n = 4). The pulps of 12 GF per broiler were each intradermally (i.d). injected with 10 μL of LPS (100 μg/mL) or 10 μL of PBS. Blood and GF were collected before (0 h) and at 6- and 24-h after GF pulp injection. Blood and GF pulp cell suspensions were immunofluorescently stained and leukocyte population profiles analyzed by flow cytometry. Blood also was used to prepare Wright-stained blood smears for differential leukocyte counts. Data were analyzed by ANOVA (P ≤ 0.05) and Fisher’s multiple means comparison tests to determine differences (P ≤ 0.05) between means. Locally, HS-LPS broilers exhibited lower levels (% pulp cells) of GF pulp infiltrating heterophils at 6- and 24-h, and lower macrophage levels at 24-h post-injection, compared to TN-LPS birds. In the blood, TN and HS broilers had similar baseline line (0 h) concentrations (cells/μL) of heterophils, monocytes, eosinophils, and basophils, but HS broilers had lower (P ≤ 0.05) total WBC and T- and B-lymphocyte levels. Concentrations of circulating heterophils and monocytes were greatly elevated (P ≤ 0.05) at 6- and 24-h, respectively, only in TN-LPS broilers, although a minimal increase (P = 0.091) in heterophils also was observed in HS-LPS broilers at 6 h. By 24 h, blood heterophils returned to pre-injection levels in both HS-LPS and TN-LPS broilers. Overall, results indicated that cyclic HS reduced both the local and systemic acute inflammatory response to LPS in broilers, likely impairing their innate defense against microbial infection. With growing concern regarding HS in the poultry industry, further research should be pursued to elucidate the mechanisms by which HS affects the innate immune system of broilers. Application of this approach may be utilized to select individuals expressing greater innate immune robustness while experiencing HS

Effects of Cyclic Heat Stress on the Acute Inflammatory Response in Broilers

Heat stress (HS) is a growing concern in broiler production due to increasing environmental temperatures. Little is known of the overall effect of HS on innate immunity. Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, is commonly used to study the inflammatory response. In avian species, the local tissue and systemic inflammatory activities in response to LPS may be determined concurrently, over time, in an individual, using the growing feather (GF) pulp dermal test along with blood measurements (dual-window approach). To examine the effect of cyclic HS on the local and systemic acute inflammatory responses, Cobb 500 male broiler chicks were reared under thermoneutral (TN) or cyclic HS conditions. Eight environmental chambers were used, four TN and four HS, with each chamber split into two pens. Beginning at 4 d of age, cyclic HS birds were subjected to 35°C from 8 am to 10 pm and TN temperatures from 10 pm to 8 am. At 37 d of age, four groups of broilers were formed: TN-LPS (n = 8 broilers), TN-PBS (phosphate-buffered saline (vehicle) control; n = 4), HS-LPS (n = 8) and HS-PBS (n = 4). The pulps of 12 GF per broiler were each intradermally (i.d). injected with 10 μL of LPS (100 μg/mL) or 10 μL of PBS. Blood and GF were collected before (0 h) and at 6- and 24-h after GF pulp injection. Blood and GF pulp cell suspensions were immunofluorescently stained and leukocyte population profiles analyzed by flow cytometry. Blood also was used to prepare Wright-stained blood smears for differential leukocyte counts. Data were analyzed by ANOVA (P ≤ 0.05) and Fisher’s multiple means comparison tests to determine differences (P ≤ 0.05) between means. Locally, HS-LPS broilers exhibited lower levels (% pulp cells) of GF pulp infiltrating heterophils at 6- and 24-h, and lower macrophage levels at 24-h post-injection, compared to TN-LPS birds. In the blood, TN and HS broilers had similar baseline line (0 h) concentrations (cells/μL) of heterophils, monocytes, eosinophils, and basophils, but HS broilers had lower (P ≤ 0.05) total WBC and T- and B-lymphocyte levels. Concentrations of circulating heterophils and monocytes were greatly elevated (P ≤ 0.05) at 6- and 24-h, respectively, only in TN-LPS broilers, although a minimal increase (P = 0.091) in heterophils also was observed in HS-LPS broilers at 6 h. By 24 h, blood heterophils returned to pre-injection levels in both HS-LPS and TN-LPS broilers. Overall, results indicated that cyclic HS reduced both the local and systemic acute inflammatory response to LPS in broilers, likely impairing their innate defense against microbial infection. With growing concern regarding HS in the poultry industry, further research should be pursued to elucidate the mechanisms by which HS affects the innate immune system of broilers. Application of this approach may be utilized to select individuals expressing greater innate immune robustness while experiencing HS

Expression of BMP-2 in vascular endothelial cells of recipient may predict delayed graft function after renal transplantation

BACKGROUND/AIMS: Delayed graft function (DGF) is associated with adverse outcomes after renal transplantation. Bone morphogenetic protein-2 (BMP-2) is involved in both endothelial function and immunological events. We compared expression of BMP-2 in epigastric artery of renal transplant recipients with immediate graft function (IGF) and DGF. ----- METHODS: 79 patients were included in this prospective study. Patients were divided in IGF group (64 patients) and DGF group (15 patients). BMP-2 expression in intima media (BMP2m) and endothelium (BMP2e) of epigastric artery was assessed by immunohistochemistry. ----- RESULTS: Lower intensity of BMP2e staining was recorded in DGF compared to IGF. In DGF patients, 93% had no expression of BMP2e and 7% had 1st grade expression, compared to 45% and 41% in IGF group, respectively (P=0.001) (P<0.001 for no expression and P = 0.015 for 1st grade expression). Patients who had BMP2e staining positive had lower odds for DGF (OR 0.059 [0.007, 0.477]) and this remained significant even after adjustment for donor and recipient variables, cold ischemia time, and immunological matching (OR 0.038 [0.003, 0.492]). ----- CONCLUSIONS: Our results demonstrate that BMP-2 expression in endothelial cells of epigastric arteries may predict development of DGF