Histopathological Changes in the Liver, Heart and Kidneys Following Malayan Pit Viper (Calloselasma rhodostoma) Envenoming and the Neutralising Effects of Hemato Polyvalent Snake Antivenom.

Calloselasma rhodostoma (Malayan pit viper) is a medically important snake species that is widely distributed across Southeast Asia. Systemic coagulopathy causing severe haemorrhage and local tissue injury is commonly observed following C. rhodostoma envenoming. However, nephrotoxicity and congestive heart failure were previously reported in a patient who had a long length of hospital stay. In this study, we determined the effect of C. rhodostoma envenoming on cardiovascular disturbances and the associated morphological changes in the liver, heart and kidneys using animal models. We also evaluated the efficacy of Hemato polyvalent antivenom (HPAV; Queen Saovabha Memorial Institute (QSMI) of the Thai Red Cross Society, Thailand) in neutralising the histopathological effects of C. rhodostoma venom. The intravenous (i.v.) administration of C. rhodostoma venom (1000 µg/kg) caused a rapid decrease in mean arterial pressure (MAP) followed by complete cardiac collapse in anaesthetized rats. Moreover, the intraperitoneal (i.p.) administration of C. rhodostoma venom (11.1 mg/kg; 3 × LD50) for 24 h caused cellular lesions in the liver and heart tissues. C. rhodostoma venom also induced nephrotoxicity, as indicated by the presence of tubular injury, interstitial vascular congestion and inflammatory infiltration in the whole area of the kidney. The administration of HPAV, at manufacturer-recommended doses, 15 min prior to or after the addition of C. rhodostoma venom reduced the extent of the morphological changes in the liver, heart and kidneys. This study found that experimental C. rhodostoma envenoming induced cardiovascular disturbances, hepatotoxicity and nephrotoxicity. We also highlighted the potential broad utility of HPAV to neutralise the histopathological effects of C. rhodostoma venom. The early delivery of antivenom appears capable of preventing envenoming outcomes.

Efficiency of Gymnema sylvestre-derived gymnemic acid on the restoration and improvement of brain vascular characteristics in diabetic rats.

The brain is a vital organ that requires a constant blood supply. Stroke occurs when the blood supply to specific parts of the brain is reduced; diabetes is an autonomous risk factor for stroke. The present study aimed to investigate the potential vascular protective effect of gymnemic acid (GM) by assessing the morphological changes of microvasculature, along with VEGFA and angiopoietin-1 (Ang-1) protein expression in the brains of diabetic rats. Rats were divided into five groups, including control, gymnemic control rats (CGM), rats that were rendered diabetic by single injection of 60 mg/kg streptozotocin (STZ), diabetic rats treated with 400 mg/kg GM (STZ + GM) and diabetic rats treated with 4 mg/kg glibenclamide (GL; STZ + GL). After 8 weeks, brain tissues were collected to examine the three-dimensional morphology of the anterior cerebral arteries by vascular corrosion casting. Western blotting was performed to determine VEGFA and Ang-1 expression. Cerebral arteries, arterioles and capillaries were depicted the diameter, thickness and collagen accumulation of the wall, and the results demonstrated narrow diameters, thickened walls and collagen accumulation in the STZ group. After receiving GM, the histopathological changes were similar to that of the control group. Through vascular corrosion casting and microscopy, signs of vessel restoration and improvement were exhibited by increased diameters, and healthy and nourished arterioles and capillaries following treatment with GM. Furthermore, VEGF expression and Ang-1 secretion decreased in the STZ + GM group compared with STZ rats. The results of the present study revealed that GM treatment decreased blood vessel damage in the brain, suggesting that it may be used as a therapeutic target for the treatment of diabetes.

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.

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.