Effect of glabridin on collagen deposition in liver and amelioration of hepatocyte destruction in diabetes rats.

Abnormalities in insulin hormone levels leads to a hyperglycemic condition of diabetic mellitus. Hyperglycemia seriously induces organ and system destructions. The excessive accumulation of collagen fiber deposits occurs in inflammatory and reorganization processes of chronic liver diseases in type I insulin-dependent diabetes. Regarding the research objective, glabridin (GLB), an active compound of licorice, was used as a daily supplement (40 mg/kg) in order to decrease hepatocyte destruction and collagen deposition in liver tissue of diabetic animals induced by streptozotocin. A total of 40 were randomly allocated to five groups (each, n=10), control, control treated with GLB (GLB), diabetic rats (DM) injected with single dose of streptozotocin (60 mg/kg) to induce a diabetic condition, diabetic rats receiving GLB (DM+GLB; 40 mg/kg) and diabetic rats treated with glibenclamide (DM+GL; 4 mg/kg). Characteristic histopathological changes in liver cells and tissues of rats were determined by Masson's trichrome staining and transmission electron microscopy (TEM). Western blotting was used to detect the expression of the key markers, collagen type I and fibronectin proteins. The histological investigation of liver tissue of the DM group revealed that the collagen fiber deposition was increased in the periportal, pericentral and perisinusoidal spaces compared with controls. Hepatocytes appeared as small and fragmented cells in TEM examination. Collagenization of the perisinusoidal space was recently demonstrated to represent a new aspect of the microvascular abnormalities and liver fibrosis. Healthy hepatocytes with round nucleus were observed following supplementation of glabridin. In addition, collagen fiber deposition was reduced in the area adjacent to the perisinusoidal space. The expression of collagen type I and fibronectin decreased strongly following glabridin supplementation in DM+GLB rats compared with DM rats, indicating that the hepatic tissue reorganization regained its normal morphology. These findings suggest that it may be beneficial to examine the role of glabridin as a therapeutic agent in diabetes treatment in future research.

Effect of Curcumin on Collagen Deposition and Tissue Reorganization in Liver of Diabetic Rats.

Objective: To localize and characterize type I and type IV collagens in recovery livers after curcumin supplementation in streptozotocin-induced diabetic rats. Material and Method: Induced diabetic rats were performed by streptozotocin injection (60 mg/kg BW). Male rats were organized into three groups, control rat (C), diabetic rat (DM) and diabetic rat supplemented with curcumin (DMC) (200 mg/kg BW). At 8 weeks, animals were sacrificed. The localization and characterization of type I and type IV collagens in liver's cell and tissues were compared among C, DM and DMC groups by Sirius red and Immunohistochemical techniques, respectively. Results: Type I and type IV collagens might be the key mediators of liver tissue healing associated with various disorders, especially with inflammation and reorganization processes. Concerning diabetic experiments, increased type I collagen was intensively recognized at subendothelial area of central veins whereas weakly demonstrated at periportal triad and perisinusoidal areas. Conversely, the high intensity of distribution of type IV collagen was strongly revealed at periportal triad and perisinusoidal areas while the intensity was faintly presented at central veins. In addition, accumulation of type IV collagen also revealed perisinusoidal basement membrane which was characteristic of capillarization of sinusoids. However, the localization of type I and type IV collagens was reduced after curcumin supplement in DMC rats compared with DM rats, implying that the liver tissue reorganization has been developed forwards to normal morphology. Moreover, type I and type IV collagen might distinctively accomplish the liver tissue reorganization by different means of area-based characterization. Conclusion: The potential beneficial effect of curcumin has been exhibited the tissue reorganization of diabetic liver tissues. The efficiency and achievement of curcumin might be applied to be an alternative therapeutic agent in diabetic hepatic pathology.

Effect of Curcumin on Characterization and Localization of Interleukin-13 and Tumor Necrosis Factor-alpha in Liver of Diabetic Rats.

Objective: To localize and characterize inflammatory markers: interleukin-13 (IL-13) and Tumor necrosis factor-alpha (TNF-alpha) in recovery livers after curcumin supplementation in streptozotocin-induced diabetic rats. Material and Method: Induced diabetic male rats were achieved by streptozotocin intravenous injection (50 mg/kg BW). The rats were divided into three groups, control rat (C), diabetic rat (DM) and diabetic rat supplemented with curcumin (DMC) (200 mg/kg BW) that has been proposed for anti-inflammation and antioxidant activities. After 12 weeks of curcumin supplementation, liver tissues were collected and processed for hematoxylin & eosin staining and immunohistochemistry. The localization and characterization of IL-13 and TNF-alpha were investigated and compared among three groups in order to analyze the efficiency of curcumin in recovering liver tissues. Results: According DM group, high intensity of IL-13 and TNF-alpha were accumulated around central vein, along hepatic parenchyma, and at perivascular sinusoidal areas. In contrast, the characterization of IL-13 and TNF-alpha in DMC were attenuated. Then, the liver tissues were recovered and engaged by less severity sign of inflammations. Therefore, dietary curcumin might have efficacy to ameliorate diabetic complications in terms of controlling and modulating inflammatory parameters, including IL-13, and TNF-alpha. Conclusion: Administration of curcumin successfully attenuated liver tissue by means of decreased inflammatory cytokine markers. The potential beneficial effects of curcumin have been shown to decline the inflammatory of liver tissue, concerning illustration of IL-13, and TNF-alpha. Therefore, the efficiency and achievement of curcumin might be applied to be an alternative therapeutic agent in diabetic liver.

Microvasculature Improvement of Heart in Diabetic Rat with Curcumin Supplementation.

OBJECTIVE: To investigate the effect of curcumin supplementation on the improvement of heart microvasculature in Streptozotocin-induced diabetic rat. MATERIAL AND METHOD: Streptozotocin (STZ: 60 mg/kg BW) was applied into rat to induce diabetic condition. Male rats were divided into three groups, control (C), diabetic (DM) and diabetic rats supplemented with curcumin (DMC) (200 mg/kg BW). After 8 and 12 weeks of experiments, heart microvasculature was investigated under vascular corrosion cast technique with scanning electron microscope (SEM). RESULTS: Destruction of heart microvasculature of DM group was observed at 8 and 12-week experiments. Five important categories of heart vessels and related veins and venules were examined respectively: right coronary arteries (RCA), medium arteries (MA), small arteries (SA), arterioles, and capillaries. RCA, cardiac arteries and veins demonstrated abnormality. Atypical patterns of vessels were presented, including shrinkage of artery vessels, capillary dropout, constriction and tortuousity of small cardiac vein and venules, and microaneurysm. At 12-week experiment, vascular lesion of DM group increased in complicated signs, including arterial constrictions and stenosis, arterial blind endings, capillary dropout and shrinkage. In addition, severity of microaneurysm dilatation of arterial branch of RCA, arterial tortuosity, coiled and twisting arteries were investigated. The diameters of vessels of all DM groups were evidently decreased. Subsequent to curcumin supplementation, typical and healthy heart microvasculatures were restored and redeveloped. The diameter sizes of DMC vessels have nearly increased back to normal situations, especially at artery, arteriole, and capillary levels. CONCLUSION: Efficiency of curcumin treatment beneficially repaired and recovered heart microvascular diabetic complications. This evidence suggests that potential anti-diabetic effect of curcumin is meaningful about the ongoing therapeutic consequences, owing to the improvement and recovery of heart blood vessels.

Effects of curcumin on restoration and improvement of microvasculature characteristic in diabetic rat's choroid of eye.

OBJECTIVE: To investigate the effect ofcurcumin on microvasculature changes in STZ-induced diabetic rat' choroid ofeye. MATERIAL AND METHOD: Male rats were divided into three groups: control (C) Diabetic rats were induced by streptozotocin (STZ) (60 mg/kg BW) (DM) diabetic rats treated with curcumin (DMC) (200 mg/kg BW). After 8 weeks of experiments, microvasculature changes of rat's choroid were studied under vascular corrosion cast technique with scanning electron microscope (SEM). RESULTS: There were pathology and destruction of choroid microvasculature of DM group that revealed reduced and shrunken sizes of large and small blood vessels, compared with control group; long posterior ciliary arteries (LPCAs) (C = 113.70 +/- 1.38, DM = 83.53 +/- 2.70, DMC = 109.64 +/- 3.41 microm), choroid arteries (C = 94.97 +/- 2.79, DM = 59.36 +/- 2.61, DMC = 80.31 +/- 3.73 microm), vortex veins (C = 74.11 +/- 3.24, DM = 46.71 +/- 2.56, DMC = 64.66 +/- 3.60 microm), and Choriocapillaris (choroidal capillaries) (C = 13.61 +/- 0.62, DM = 4.46 +/- 0.24, DMC = 9.96 +/- 0.70 microm), respectively. In DM group, LPCAs and Choroid arteries were tortuous and showed shrinkage. Vortex veins became narrow. Choriocapillaris showed the pathological characteristics of vascular lesions including of shrinkage, constriction, microaneurysm and blind ending. Fascinatingly, Choroid microvasculature of the eye in curcumin treated group developed into regenerate and repaired conditions with healthy and normal characteristics. CONCLUSION: Efficiency of curcumin treatment beneficially repaired and regenerated the redevelopment of choroid's microvascular complications of eye in 8-week experiments. Potential treatment with curcumin in diabetes has demonstrated in a meaningful way the therapeutic consequences in the improvement and recovery of choroidal blood vessels in eye pathology ofdiabetic rats.

Study of curcumin on microvasculature characteristic in diabetic rat's liver as revealed by vascular corrosion cast/scanning electron microscope (SEM) technique.

OBJECTIVE: To investigate the effect of curcumin on the structural change ofmicrovasculature in STZ-induced diabetic rat' liver. MATERIAL AND METHOD: Diabetic rats were induced by streptozotocin (60 mg/kg BW). Male rats were divided into thre groups, control (C), diabetic (DM) and diabetic rats treated with curcumin (DMC) (200 mg/kg BW). After 8 weeks o experiments, blood vessels of rat's liver were studied under conventional light microscope (LM) and vascular corrosion cas technique with scanning electron microscope (SEM). RESULTS: LM observation demonstrated that there were pathology and destruction of liver tissues and microvasculature in diabetic animals. The sinusoids around central veins were dilated and filled with red blood cells. There was an accumulation of lipid droplets in the cytoplasm of hepatocytes and hepatocyte nuclei showed pathological sign of pyknosis. Moreover, the inflammation change of liver tissues revealed the infiltration of lymphocytes and increasing of collagen deposition in the area of portal triad. In curcumin-treated rats, the distinguished recovery of liver tissues showed regained normal pattern of central veins, sinusoids, hepatocytes and portal triad, when compared with liver tissues of control group. By using vascular corrosion casting with SEM, the liver blood vessels of DM group revealed higher and expanded sizes, compared with control group; proximal parts of portal veins (C = 577.75 +/- 126.23, DM = 892 +/- 35.79, DMC = 469.5 +/- 8553 microm), distal parts of portal veins (C = 76.72 +/- 1.48, DM = 200 +/- 31.05, DMC = 76.38 +/- 2.98 microm) and venules (C = 27.03 +/- 0.55, DM = 45.15 +/- 5.03, DMC = 28.38 +/- 3.67 microm) and corresponding to increased blood volumes compared with control group; proximal parts of portal veins (C = 20.8 +/- 1.28, DM = 62.2 +/- 3.39, DMC = 14.9 +/- 0.67 microm3), distal parts of portal veins (C = 0.46 +/- 0.03, DM = 3.81 +/- 0.18, DMC = 0.41 +/- 0.05 microm3) and venules (C = 0.05 +/- 0.05, DM = 0.24 +/- 0.013, DMC = 0.05 +/- 0.05 microm3) respectively. Fascinatingly, liver microvasculature in curcumin treated group developed into regenerate and repair into healthy and normal characteristics. CONCLUSION: Efficiency of curcumin treatment beneficially repaired and regenerated liver tissues of diabetic groups and also redeveloped the liver's microvascular complications. These results optimistically demonstrated the potential use of curcumin as a novel therapeutic agent in liver pathology of diabetic rats.

Maturation of the gastric microvasculature in Xenopus laevis (Lissamphibia, Anura) occurs at the transition from the herbivorous to the carnivorous lifestyle, predominantly by intussuceptive microvascular growth (IMG): a scanning electron microscope study of microvascular corrosion casts and correlative light microscopy.

The microvascular bed of the stomach of Xenopus laevis and the changes it undergoes when the herbivorous tadpole becomes a carnivorous adult were studied by scanning electron microscopy of vascular corrosion casts and light microscopy of stained tissue sections. In tadpoles an upper and a lower gastric artery supplied, and upper, middle and lower medial and lateral gastric veins drained the vertically extending stomach. During metamorphosis, the stomach gained a horizontal cranio-caudal extension and vessels accordingly become dorsal and ventral gastric arteries, and anterior, middle and posterior gastric veins, respectively. Up to stage 64 (late climax) mucosal capillaries formed a polygonal network of wide immature-looking capillaries ensheathing gastric glands in a basket-like manner. From stage 64 onwards, blood vessels of the stomach appeared mature, revealed a clear hierarchy and were correlated closely with the histomorphology of the stomach, which had also gained the adult pattern. Within the gastric mucosa, ascending arterioles branched in a fountain-like pattern into wide subepithelial capillaries establishing a centripetal blood flow along the gastric glands, which makes an ultrashort control loop of glandular cells within the branched tubular gastric glands very unlikely. Formation of the stomach external muscular layer started at stage 57 when smooth muscle cells locally formed a single longitudinal and one-to-two single circular layers. Abundant signs of intussusceptive microvascular growth and rare vascular sprouts in vascular corrosion casts indicated that the larval-to-adult microvascular pattern formation of the stomach of Xenopus laevis Daudin occurs predominantly by non-sprouting angiogenesis.

Effect of Vernonia cinerea in improvement of respiratory tissue in chronic nicotine treatment.

OBJECTIVE: To demonstrate the effect of Vernonia cinerea (VC) on rat respiratory tissue in chronic nicotine condition. MATERIAL AND METHOD: Pathology of rat respiratory tissue was induced by intraperitoneally injection with 1 mg/kg BW of rat. Male Wistar rats were divided into three groups, control group (C), nicotine treated group (N) and nicotine treated with Vernonia cinerea (VC) supplementation (NV, 100 mg/kg BW of rat) for 3 and 6 months. The animals were sacrificed and the respiratory tissues were removed and further processed for paraffin embedment and stained with Hematoxylin & Eosin (H&E), Periodic Acid Schiff (PAS), and Masson's trichrome techniques. RESULTS: The histopathology of lung tissue and trachea occurred in a chronic nicotine treatment. The thickness of alveolar walls and proliferation of alveolar type 2 cell were found. There was remarkable increasing of various inflammatory cells, alveolar macrophages, lymphocytes and plasma cells after nicotine treatment for 6 months. A large number of small blood vessels appeared in the alveolar wall. Nicotine also caused fibrosis which dispersed throughout the lung parenchyma in perivascular peribronchiole and alveolar wall regions. Moreover there was the appearance of epithelial cell injury and goblet cell hyperplasia in the trachea. Regarding the VC supplementation, the result of a recovery of alveolar walls, i.e. decreasing of various inflammatory cells and alveolar type 2 cells was clearly demonstrated. In addition, the fibrosis and goblet cell hyperplasia were almost disappeared in the lung tissue after VC treatment. CONCLUSION: Administration of VC in a chronic nicotine treatment resulted in an improvement of respiratory tissue. The recovery of the respiratory tract, especially trachea and lung tissue was characterized by the remarkable decrease of various inflammatory cells, fibrotic areas, and goblet cell hyperplasia. The VC, therefore shows the potential effect to be a new herbal therapeutic agent for alleviate the symptoms of the respiratory tract caused by nicotine from heavy cigarette smoke.

Paramphistomum cervi: surface topography of the tegument of adult fluke.

Adult Paramphistomum cervi or rumen fluke are pear-shaped, slightly concave ventrally and convex dorsally. The worm measures about 5-13 mm in length and 2-5 mm in width across the mid-section. As observed by scanning electron microscopy (SEM), the tegumental surface in all part of the body, appears highly corrugated with transverse folds alternating with grooves and is spineless. At high magnification, the surface of the fold is composed of microfolds or ridges separated by microgrooves or pits. Corrugations and invaginations of the ventral surface are also more extensive than on the dorsal surface of the body. Both anterior and posterior suckers have thick rims covered with transverse folds without spine. The genital pore is situated at the anterior third of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape, measuring 10-15 microm in diameter at the base with nipple-like tips, and type 2 has a similar shape and size and also a short cilia on top. These sensory papillae usually occur in large clusters, each having between 5 and 20 units depending on the region of the body. Clusters of papillae on the ventral surface and around the anterior suckers tend to be more numerous and larger in size. The dorsal surface of the body has the least number of papillae.

Effect of curcumin in the amelioration of pancreatic islets in streptozotocin-induced diabetic mice.

OBJECTIVE: To evaluate the antidiabetic effect of curcumin and its potential in amelioration of pancreatic islets against damage under diabetic condition. MATERIAL AND METHOD: Diabetic mice were induced by injection of STZ (60 mg/kg body weight). Male mice were divided into 3 groups: group I was normal mice, group II was diabetic mice and group III diabetic mice were treated with curcumin (200 mg/kg body weight). The blood glucose levels and body weights were recorded every two weeks. After 4 weeks, 8 weeks and 12 weeks, the animals in each group were sacrificed. Histopathology of pancreatic tissues, pancreatic islets areas and numbers were observed under light microscope. RESULTS: The weight loss and the elevation of blood glucose levels were observed in diabetic mice and diabetic mice treated with curcumin at 4 weeks, 8 weeks and 12 weeks. The reduction of pancreatic islets areas and numbers were presented in diabetic mice and diabetic mice fed with curcumin at 4 weeks. At 8 weeks of diabetic mice, the numbers of pancreatic islets were decreased however the pancreatic islets hyperplasia was prominently investigated, whereas the noticeable increase in numbers of small pancreatic islets were observed in diabetic mice fed with curcumin. Histopathological observation at 12 weeks revealed the accumulation of lymphocytes in the shrunken pancreatic islets of diabetic mice, while an absent lymphocytes infiltration in the pancreatic islets and the increase in numbers of small islets of Langerhans appeared nearly the ducts in the pancreas of diabetic mice treated with curcumin at 12 weeks. CONCLUSION: Curcumin treatment at 12 week can exert beneficial effect in diabetes mellitus, regarding the improvement of pancreatic islets. The islets of Langerhans neogenesis is characterize by the presentation of small islets increase in numbers nearly the ducts and no insulitis.

Effect of curcumin on vascular endothelial growth factor expression in diabetic mice kidney induced by streptozotocin.

OBJECTIVE: To localize and demonstrate the effect of curcumin on vascular endothelial growth factor in diabetic mice kidney induced by streptozotocin. MATERIAL AND METHOD: Diabetic mice were induced by streptozotocin (60 mg/kg BW). Male mice were divided into three groups, control mice, diabetic mice (DM) and diabetic mice treated with curcumin (DMC) (200 mg/kg BW). At 4 and 8 weeks, animals were sacrificed and kidneys were processed by immunohistochemistry technique. RESULTS: At the end of 4 and 8 week experiments, glomeruli were slightly enlarged and showed diffuse thickening of the glomerular capillary walls in diabetic mice. Administration with curcumin presented the better improvement and recovery of cells and tissues compared with diabetic mice. Immunohistochemical staining for vascular endothelial growth factor (VEGF) demonstrated that VEGF was mainly detected in the podocytes and renal tubules. There was an increase in VEGF expression in diabetic mice as compared to control. Treatment with curcumin significantly inhibited the expression of VEGF in the kidney tissue of diabetic mice in both 4 and 8 weeks. Comparing the diabetic mice between 4 and 8 week experiments, the expression of VEGF in the podocytes and renal tubules at 8 week were significantly stronger than at 4 week which represented time-dependent change. Nevertheless, the intensity of VEGF was not different in DMC mice when it was compared between 4 and 8 weeks. CONCLUSION: VEGF immunoreactivity of the podocytes and the renal tubules at 4 and 8 weeks in DM mice showed strong intensity more than in control mice. However, the intensity of VEGF in DMC mice was less when it was compared with DM mice. Moreover, VEGF was a key modulator of angiogenesis and a potent mitogen for endothelial cells. These results demonstrated the potential use of antiangiogenic curcumin as a novel therapeutic agent in diabetic mellitus and maintain normal structure of the kidney.

Origins and pathways of fluid entering sublobular lymphatic vessels in cat livers.

The liver, which produces a large volume of lymph, has a lymphatic system which can be classified into three categories: portal, sublobular, and superficial lymphatic vessels. As little is known about the origin and pathways of sublobular lymph, this study demonstrates pathways of interstitial fluid flowing into sublobular lymphatic vessels. Livers from cats whose thoracic ducts were either ligated or non-ligated were examined by light-, transmission electron- and scanning electron-microscopy (SEM). Complete ligation of the thoracic duct caused significant dilation of the hepatic sinusoids, the space of Disse, and channels passing through the limiting plate. Sublobular interstitial space and sublobular lymphatic vessels were also expanded. The channels between hepatocytes forming the limiting plate contained collagen fibers, and connected the space of Disse with a sublobular interstitial space. The alkali-water maceration/SEM confirmed that collagen fibers traversing the layer of the limiting plate independently of blood vessels connected collagen fibers in the space of Disse with those in the sublobular space. Complete ligation of the thoracic duct also showed an accumulation of mast cells and plasma cells in the sublobular interstitial space. Our data suggest that fluid in the space of Disse flows along collagen fibers in channels traversing the limiting plate as well as those along the sinusoids and central veins that drain into sublobular veins, and enters the sublobular interstitial space to finally lead into sublobular lymphatic vessels. Our study has also shown that hepatic lymphostasis causes the accumulation of mast cells and plasma cells in the sublobular interstitial space, which may be involved in lymphangiogenesis and fibrogenesis.

Scanning electrochemical microscopy of model neurons: imaging and real-time detection of morphological changes.

Living PC12 cells, a model cell type for studying neuronal function, were imaged using the negative feedback mode of a scanning electrochemical microscope (SECM). Six biocompatible redox mediators were successfully identified from a large pool of candidates and were then used for imaging PC12 cells before and after exposure to nerve growth factor (NGF). When exposed to NGF, cells differentiate into a neuron phenotype by growing narrow neurites (1-2 microm wide) that can extend > 100 microm from the cell proper. We demonstrate that carbon fiber electrodes with reduced tip diameters can be used for imaging both the cell proper and these neurites. Regions of decreased current, possibly resulting from raised features not identifiable by light microscopy, are clearly evident in the SECM images. Changes in the morphology of undifferentiated PC12 cells could be detected in real time with the SECM. After exposure to hypotonic and hypertonic solutions, reversible changes in cell height of <2 microm were measured.

Effects of ozone treatment on cell growth and ultrastructural changes in bacteria.

Ozone appeared to inhibit growth and caused the death of gram negative and gram positive tested bacteria: Escherichia coli, Salmonella sp., Staphylococcus aureus and Bacillus subtilis. Bacterial cultures at 10(3), 10(4), 10(5), 10(6), and 10(7) cfu/ml dilution were exposed to 0.167/mg/min/L of ozone at different time intervals (0, 5, 10, 15, 30, 60, 90, 120, and 150 min). Cell viability was observed in all types of tested bacteria at 10(3), 10(4), 10(3) cfu/ml within 30 min after ozone exposure. However, cell inactivation was not significantly observed at concentrations of 10(6), 10(7) cfu/ml even after an exposure of 150 min. Ultrastructural changes of treated bacteria showed deformation, rough damage and surface destruction revealed by scanning electron microscopy. Some bacterial cells showed collapsed and shrunken patterns within 60 min and severe rupture and cellular lysis after 90 min of ozone treatment. This study supports the proposed mechanism of the bacteria inactivation by ozone that caused cell membrane destruction and finally lysis reaction. Thus, the precaution of using ozone as a biocide should be used to address appropriate concentrations of bacterial contamination in water.