Preventive effects of cannabis on neurotoxic and hepatotoxic activities of malathion in rat

Objective:To investigate the effect of Cannabis sativa extract on the development of neuro- and hepato-toxicity caused by malathion injection in rats.Methods:The extract of Cannabis sativa was obtained from the plant resin by chloroform treatment. Δ-Tetrahydrocannabinol content of the extract (20%) was quantified using gas chromatography–mass spectrometry. The doses of cannabis extract were expressed as Δ -tetrahydrocannabinol content of 10 or 20 mg/kg. Malathion (150 mg/kg) was intraperitoneally administered followed after 30 min by the cannabis extract (10 or 20 mg/kg, subcutaneously). Rats were euthanized 4 h later. Malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide and paraoxonase-1 (PON-1) activity were determined in brain and liver. Brain 5-lipoxygenase and butyrylcholinesterase (BChE) activity were measured as well. Histopathological examination of brain and liver tissue was also performed.Results:Compared to controls, malathion resulted in increased oxidative stress in brain and liver. MDA and nitric oxide concentrations were significantly increased (P<0.05) and GSH significantly decreased with respect to control levels (P<0.05). Malathion also significantly inhibited PON-1 and BChE activities but had no effect on brain 5-lipoxygenase. Brain MDA concentrations were not altered by cannabis treatment. Cannabis at 20 mg/kg, however, caused significant increase in nitric oxide and restored the GSH and PON-1 activity. Brain BChE activity significantly decreased by 26.1% (P<0.05) after treatment with 10 mg/kg cannabis. Cannabis showed no effect on brain 5-lipoxygenase. On the other hand, rats treated with cannabis exhibited significantly higher levels of liver MDA, nitric oxide and PON-1 activity compared with the malathion control group. Rats treated with only malathion exhibited spongiform changes, neuronal damage in the cerebral cortex and degeneration of some Purkinje cells in the cerebellum. There were also hepatic vacuolar degeneration and dilated and congested portal vein. These histopthological changes induced by malathion in brain and liver were reduced to great extent by cannabis administration at 20 mg/kg.Conclusions:Our data suggest that acute treatment with cannabis alleviates the malathion-induced brain and hepatic injury in rats possibly by maintaining the levels of GSH and PON-1 activity.

Protective effect of fish liver oil and propolis on anticonvulsant drugs-induced osteoporosis

Osteoporosis is a major health problem and its prevalence increases the risk of bone fracture. It is classified into primary [postmenopausal or age related] and secondary [related to chronic diseases, drug therapy, or life style]. There is accumulating evidence that patients on antiepileptic drugs [AEDs] are at an increasing risk of developing osteoporosis. The present study aimed at investigating the protective effect of dietary natural products, fish liver oil, and propolis on osteoporosis caused by anticonvulsant drugs. A total of 105 albino rats were used, divided into seven groups of 15 rats each. Group 1 was used as a control group. In group 2, rats were injected intraperitoneally with pilocarpine [300 mg/kg body weight]. The pilocarpine-induced epileptic rats in the other five groups were orally treated with valproate [400 mg/kg body weight], a combination of valproate and fish liver oil [0.4 ml/kg body weight/day], a combination of valproate and propolis [50 mg/kg body weight/ day], fish liver oil, and propolis, respectively. At the end of the experiment [6 months treatment], animals were sacrificed, femur shafts were extracted, decalcified, and processed into paraffin blocks for histopathological and image analysis and morphometric studies. Rats treated with the antiepileptic valproate alone showed a decrease in the thickness of shaft cortical bone, with a marked decrease in the number of osteocytes, increase in Haversian canals, and decrease in bone trabeculae, disruption of normal architecture, and widening of bone marrow spaces compared with the control group. Treatment with the dietary natural products, fish liver oil, and propolis along with the AED valproate might improve histopathological changes and morphometric parameters in bone associated with AED-induced osteoporosis

Vitamin C and E antagonistic effect for tamoxifen-induced rat liver damage: a histopathological, histochemical and ultrastructural studies

Tamoxifen is widely used to treat oestrogen dependent carcinoma of the breast. Previous long term studies have shown that oral administration of tamoxifen induces hepatoproliferative lesions and hepatocellular tumors in rats. This study was designed to evaluate the effects of tamoxifen on liver of rats and the possible protective effects of vitamin C and/ or vitamin E in amelioration of these effects. A total of 70 adult female albino rats were used in this study. The animals were divided into seven groups. Each group contained 10 rats. The rats of the first group were kept as control. Animals of the second group were daily dosed orally with tamoxifen 20 mg/ kg b. w. by stomach tube for two weeks. The third group was given vitamin C at dose level of 0.01 g/ 100 g b w by stomach tube, 15 min before daily administration of tamoxifen. The fourth group was given vitamin E at dose level of 100 mg/ kg b.w, 15 min prior to daily administration of tamoxifen throughout the whole period. The fifth group was given combination of the two vitamins [vitamin C and vitamin E] at dose level of 0.01 g/ 100 gb.w. and 100 mg/ kg b.w. respectively, 15 min before daily administration of tamoxifen for two weeks. Each of the remaining two groups was daily given vitamin C [0.01 g/ 100 g b.w.] and/ or vitamin E [100 mg/ kg b.w.] only for two weeks. Paraffin sections were used for the histopathological study. For the histochemical investigations, sections were stained to demonstrate DNA, mucopolysaccharides and protein content. Histopathological effects of tamoxifen were demonstrated in liver as vacuolar degeneration, fatty changes and hydropic degeneration. Signs of degeneration in the form of karyolysis and karyorrhexis were also seen. Moderate dilatation of blood sinusoids, some dysplastic cells and chromalin clumping could be observed. Quantitative DNA image analysis [Lecia image] showed a decrease in DNA content [hypoploidy] in liver of rats treated with tamoxifen only. Tamoxifen induced histochemical changes consisted of marked diminution of protein and mucopolysaccharides content. No histopathological, histochemical and ultra structural changes could be noticed in rats treated with vitamin C and, or vitamin E only. The treatment of rats with vitamin C and/ or vitamin E prior to tamoxifen resulted in amelioration of the histopathological changes of liver as well as histochemical and ultrastructural changes

Effect of Trimebutine on hepatic injury caused by carbon tetrachloride in the rat

We investigated the effects of trimebutine, a peripheral opiate agonist, used in therapy of irritable bowel syndrome on the development of hepatic injury in rats treated with carbon tetrachloride. When administred s.c., at 30, 60 and 120 mg/kg, the drug increased the degree of hepatic damage caused by CCL[4] in a dose-dependent manner as reflected by serum alanine aminotransferase [ALT], aspartate aminotransferase [AST], alkaline phosphatase [ALP] and liver histopathology determined 14 days after drug administration. The effect was dose-dependant one, being most evident after administration of 60 and 120 mg/kg dose of the drug. Thus, compared with the CCl[4][control group, serum ALT increased by 28.9, 47.2%, AST by 16.2, 24.8% and ALP by 28.8, 47.2% after the administration of trimebutine at 60 and 120 mg/kg, respectively. When silymarin was combined with 120 mg/kg of trimebutine, the activities of ALT and AST were comparable with those seen after the administration of silymarin alone, suggesting a protective effect for silymarin against the increases in tranaminases by trimebutine. Histopathologic examination of the liver of rats treated with CCl[4] + trimebutine at a dose of 120 mg/kg showed massive inflammatory cellular infiltrate, fatty change and vacuolar degeneration. These changes were less marked in rats treated with CCl[4] + silymarin + 120 mg/kg trimebutine. It is concluded that the opiate agonist trimebutine aggrevates hepatocellular injury caused by CCl[4] in rats, which can be lessened by coadminisration of silymarin. We suggest monitoring of hepatic functions in patients on long term use of this drug, especially in those with already known hepatic dysfunction such as steatosis or HCV infected individuals. The administration of silymarin in patients on long-term use of trimebutine is advisable

Effect of the new anti-ischaemic drug "trimetazidine" on hepatic injury caused by carbon tetrachloride in rats

The effects of trimetazidine, a novel anti-ischaemic agent, on the development of hepatic injury induced in rats with carbon tetrachloride, were investigated. Hepatotoxicity was induced by CCl[4] orally [0.2 ml/kg followed by 0.1 ml/kg after one week]. Trimetazidine at three dose levels [3.6, 7.2 and 14.4 mg/kg], silymarin [25 mg/kg] and combination of trimetazidine [7.2 mg/kg] and silymarin [25 mg/kg] were administered orally daily for 15 days, starting at time of administration of CCL. The daily administration of trimetazidine conferred significant protection against the hepatotoxic effects of CCL, in rats. It decreased the increases in serum alanine aminotransferase [ALT], aspartate aminotransferase [AST], and alkaline phosphatase [ALP] and also prevented the development of hepatic necrosis caused by CCL as determined 15 days after drug administration. Thus, compared with the CCL control group, serum ALT decreased by 35.1, 49.5 and 57%, AST by 28.2, 31.6 and 36% and ALP by 32, 39.7 and 40.6%, after the administration of trimetazidine at 3.6, 7.2 and 14.4 mg/kg, respectively. Silymarin administered alone, at the dose of 25 mg/kg reduced serum ALT by 30%, AST by 29.1% and ALP by 38.4% compared to CCL control. When silymarin was combined with 7.2 mg/kg of trimetazidine, the activities of ALT, AST and ALP were markedly decreased by 47.2%, 47.2% and 50.6%, respectively, compared with the CCl[4]control, indicating a beneficial additive effect. Histopathologic examination of the liver of rats treated with CCl[4] + trimetazidine showed less fibrosis compared with the CCl[4] -control group. Co-treatment with silymarin and trimetazidine, on the other hand resulted in marked histologic protection. It is concluded that the anti-ischaemic agent trimetazidine lessened hepatocellular injury caused by CCl[4], in rats, and had additive effect with silymarin. It was suggested, therefore, that trimetazidine alone or in combination with silymarin might have a place in the therapy of chronic liver diseases

Ductal breast tumours: mean nuclear area and metallothionein expression in correlation with estrogen receptor status

Estrogen is an important growth Ilictor for breast tumour playing an important role in regulating the proliferation and differentiation of normal and malignant mammary epithelial cells. Nuclear morphometry and Metallothioneins [MTs] are indicators of proliferation that have been used as predictors of prognosis in many tumours. The present study aimed to study mean nuclear area and MT; ER expression in fibroadenoma, ductal carcinoma in situ and infiltrating ductal carcinoma of the breast. Also MNA and MT expression will be correlated with histologic grade and ER status in breast carcinoma. Breast tissues from 12 patients with fibroadenoma [FA], 6 patients with ductal carcinoma in situ [DCIS] and 20 patients with infiltrating durtal carcinoma [IDC] were used in this study. Mean nuclear area [MNA] and metallothionein [MT] expression; as proliferation markers; were investigated and correlated with estrogen receptor [ER] status. All cases of fibroadenoma, 4 out of 6 cases [66.7%] of DCIS and 12 out of 20 cases [60%] of IDC were positive for ER. MNA of cancer cells was significantly larger than that of normal and benign breast tissue. A significant direct correlation was found between MNA and histologic grades. MNA of ER negative carcinomas was significantly larger than that of ER positive tumours. In normal and benign breast tissue, myoepithelial cells consistently expressed the MT protein. Two out of 6 DCIS cases [33.3%] and 13 out of 20 cases of IDC cases [65%] were positively stained for MT. MT positivity was directly correlated with histologic grade of JDC. There was a highly significant inverse correlation between MT and ER over-expression. From this study, it is concluded that in invasive ductal carcinoma of the breast, the large MNA and MT over-expression are correlated with histologic grades and ER negativity. Therefore, MNA and MT overe-expression may be possible important indicators for more aggressive and less differentiated breast carcinoma

protective role of ginseng against liver damage caused by potassium dichromate Cr [IV]: light and electron microscopic studies

Human exposure to carcinogenic Hexavalent Chromium Cr [VI] compounds is found among workers in a large number of professional groups, and it can also occur through environmental pollution. A significant number of toxic waste sites contain Cr as a major contaminant. Chromium is a heavy metal which is widely used in the painting, pigments, dyes, and leather tanning industries. Ginseng is one of the most popular herbal remedies. In a series of experiments we have studied the possible role of the water ginseng extract, in modulating the histological and histochemical damage induced by potassium dichromate Cr [VI] in liver tissue of albino rats. Fourty male albino rats were used in this study. Potassuim dichromate was given orally in a dose of 50mg/kg body weight. Rats were treated orally with ginseng extract in a dose of 20 mg/kg body weight. Specimens of the liver were taken and prepared for light and electron microscopic investigations. Other specimens were stained with, periodic-shiff reagent and Feulgen stain for histochemical investigations. The morphometric analysis of hepatocytes nuclear diameters, the optical density of mucopolysaccharide and DNA reactions were evaluated by the computer image analysis. Light microscopic investigation of liver of rats treated with Cr [VI] showed massive vacuolar degeneration and circumscribed nodule formed of a homogenous acidophilic material infiltrated by mononuclear cells. Apoptic cells with deep eosinophilic cytoplasm and small deeply stained [pyknotic] or fragmented nuclei were clearly demonstrated together with fatty degeneration manifested by round empty well-circumscribed spaces. Necrosis of hepatocytes was observed, they showed irregular nuclei densly packed with chromatin. At the ultrastructural level the hepatocytes revealed that endoplasmic reticulum was slightly swollen and vesiculated. In addition, different degrees of mitochondrial damage were present in the form of differences in shape and size and distortion of their ridges. The results have manifested a possible protective role of ginseng extract on the general cellular morphology, as well as significant improvement in glycogen and DNA contents in ginseng treated rats against chromium toxicity

Hepatorotective effect of garlic [Allium sativum] and vitamin E on experimental liver injuries induced by ultraviolet radiation

Previous researches have demonstrated that the garlic and vitamin E were able to exert preventive properties against sunburn, delay the onset of skin tumors and reduce radiation induced tissue damage. These compounds may act as antioxidants able to scavenge free radicals and lipid peroxidation. The aim of the present study is to investigate the possible protective effect of garlic and vitamin E against ultraviolet radiation induced liver damage. A total of 60 male albino rats weighing 180-200 g were used in this study. The rats were divided into six groups. Each group contained 10 rats. Group [1] Animals were kept as control. Group [2] Animals were exposed to ultraviolet C-rays 180-280 nm for 30 successive days. Group [3] was given ethanolic extract of garlic at dose level of 0.18 ml/100 g.15 min before exposure to ultraviolet C-rays 180-280 nm for 30 successive days. Group [4] was given vitamin E at dose level of 100 mg/kg b. w 15 min before exposure to ultraviolet C-rays 180-280 nm. The fifth and sixth groups were given garlic and vitamin E respectively for 30 successive days. Histopathological effects in liver were demonstrated as necrosis, fibrosis, fatty changes, inflammatory cellular infiltration and vacuolar degeneration. Deep nuclear basophilia and karyolysis were also seen in some hepatocytes. No pathological, histochemical and ultrastructural changes could be observed in rats treated with garlic or vitamin E. Histochemical results showed marked diminution of glycogen content, DNA, protein content and increase in collagen deposition. Ultrastructural changes were observed in irradiated rats in the form of areas of cytoplasmic dissolution, partial clumping of nuclear chromatin and partial disappearance of nuclear membrane. Mitochondria had dense matrix with proliferation and vesiculation of endoplasmic reticulum. The treatment of rats with ethanolic extract of garlic or vitamin E before exposure to ultraviolet C-rays alleviated the deleterious effects of ultraviolet radiation

Antioxidative effects of melatonin in protection against renal tubular damage caused by fumonisin [histological, histochemical and electronmicroscopical studies]

Melatonin is the chief secretory product of the pineal gland and has a very potent antioxidant activity. This study was designed to evaluate the effects of the mycotoxin fumonisin on renal tubular damage and protective role of melatonin to reduce this effect. A total of 48 female albino rats weighing [100-140 g] were used in this study. The animals were divided into six groups, each group contained 8 rats. The first group served as control. The animals of the second group were fed ration contaminated with fumonisin [100 mg/kg diet]. The third group of rats were fed ration contaminated with fumonisin [200 mg/ kg diet]. The fourth group was given daily interperitoneal injection of 10 mg/ kg melatonin and fed ration contaminated with fumonism [100 mg/ Kg diet]. The fifth group was given daily interperitoneal injection of 10 mg/kg melatonin and fed ration contaminated with fumonisin [200 mg/kg diet]. The sixth group was given melatonin only at a dose level of 10 mg /kg. The rats were treated for one month Histological effects in kidney were demonstrated in the form of vacuolar degeneration in tubular epithelial cells with degenerated and pyknotic nuclei in animals fed ration contaminated with fumonisin [100 mg/kg diet], markedly degenerated tubules and glomerular degeneration were noticed in group of rats treated with fumonisin [200 mg/kg diet]. Histochemical results showed marked diminution of protein content, mucopolysaccharides and increase in alkaline phosphatase activity. Quantitative DNA image analysis [Leica image] showed that the kidney contained 12.15% aneuploid cells in the group of rats fed fumonisine at dose level of 200 mg/kg diet. No pathological, histochemical or ultrastructural changes could be noticed in rats treated with melatonin only. Ultrastructural changes were observed in animals fed ration contaminated with fumonisin in the form of shrinkage and degeneration of nuclei. The mitochondria were condensed, fragmented and variable in size and shape. The endoplasmic reticulum was fragmented. The treatment of rats with melatonin along with fumonisin resulted in an improvement in histolgical picture as well as histochemical parameters and ultrastructural changes