Effect of crocin, an active saffron constituent, on ethanol toxicity in the rat: histopathological and biochemical studies.

OBJECTIVES: The drinking of ethanol causes the wide range of clinical illness and morphological changes including hepatotoxicity and nephrotoxicity. In the current study, the protective properties of crocin versus oxidative stress, apoptosis and inflammation induced by ethanol were assessed. MATERIALS AND METHODS: The male Wistar rats were divided into seven groups consisting of 6 rats in control, ethanol (50% v/v - 5 g/kg), crocin (10, 20 and 40 mg/Kg) plus ethanol, crocin 20 and 40 mg/Kg. RESULTS: The MDA level was remarkably enhanced, while the content of GSH was significantly diminished in the kidney and liver of alcoholic rat but protective groups restored the level of MDA and GSH contents. Ethanol consumption induced hepatotoxicity and nephrotoxicity as evidenced by biochemical abnormalities and histopathological damages but crocin improved them. Also, crocin restored the TNF-α and IL-6 levels in the liver. The consumption of ethanol enhanced the levels of caspase-3, -8, -9 and Bax/Bcl-2 ratio (mRNA and protein) but, western blot and real-time PCR data confirmed that crocin treatment prevented apoptosis induced by ethanol. CONCLUSION: This research demonstrates that crocin has protective activities against ethanol toxic effects in rat liver and kidney via anti-inflammatory, antioxidant, and anti-apoptotic effects.

Effect of saffron (stigma of Crocus sativus L.) aqueous extract on ethanol toxicity in rats: A biochemical, histopathological and molecular study.

ETHNOPHARMACOLOGICAL RELEVANCE: Saffron (Crocus sativus L.) is considered in the Iranian traditional medicine because of many therapeutic properties such as sedative agent, strengthen the stomach and liver, improving the uterus disorders and infectious wounds. The detoxification of alcohol was one of the most important of saffron effects in ancient medicine. AIM OF THE STUDY: In the current research, the protective effects of saffron aqueous extract (Aq. Ext.) versus oxidative stress, apoptosis, inflammation, histopathological and biochemical abnormalities induced by ethanol were evaluated. MATERIALS & METHODS: The male Wistar rats were divided into seven groups consisted of 6 rats in control (distilled water), ethanol (5 g/kg - 50% v/v), Aq. Ext. (40, 80 and 160 mg/kg) plus ethanol, Aq. Ext. 80 and 160 mg/kg. Animals were treated for four weeks and at the end of treatment period, histopathological damages, biochemical markers, apoptosis, levels of MDA and GSH, TNF-α and IL-6 were evaluated. RESULTS: Ethanol induced nephrotoxicity and hepatotoxicity as evidenced by histopathological damages and biochemical abnormalities. The level of MDA was significantly enhanced while GSH content was remarkably reduced in ethanol-treated rats, but protective groups restored them. Also, the levels of TNF-α and IL-6 were regulated by Aq. Ext. Furthermore, the effects of ethanol on histopathological and biochemical parameters were improved by Aq. Ext. The ethanol increased the expression of Bax/Bcl2 ratio, caspase-3, -8, and -9. Real-time PCR and western blot analysis proved that Aq. Ext. treatment inhibited apoptosis induced by ethanol through decreasing the Bax/Bcl2 ratio (mRNA and protein) and caspases-3, -8, and -9 levels in the kidney and liver. CONCLUSION: The results of this research demonstrated that Aq. Ext. could exert protective effects against ethanol toxicity in rat kidney and liver via antioxidant, anti-apoptosis, and anti-inflammatory effects.

Protective effect of thymoquinone, the active constituent of Nigella sativa fixed oil, against ethanol toxicity in rats.

OBJECTIVES: Long term consumption of ethanol may induce damage to many organs. Ethanol induces its noxious effects through reactive oxygen species production, and lipid peroxidation and apoptosis induction in different tissues and cell types. Previous experiments have indicated the antioxidant characteristics of thymoquinone, the active constituent of Nigella sativa fixed oil, against biologically dangerous reactive oxygen species. This experiment was planned to evaluate the protective effect of thymoquinone against subchronic ethanol toxicity in rats. MATERIALS AND METHODS: Experiments were performed on six groups. Each group consisted of six animals, including control group (saline, gavage), ethanol-receiving group (3 g/kg/day, gavage), thymoquinone (2.5, 5, 10 mg/Kg/day, intraperitoneally (IP)) plus ethanol and thymoquinone (10 mg/Kg/day, IP) groups. Treatments were carried out in four weeks. RESULTS: Thymoquinone reduced the ethanol-induced increase in the lipid peroxidation and severity of histopathological alteration in liver and kidney tissues. In addition it improved the levels of proinflammatory cytokines in liver tissue. Furthermore, thymoquinone corrected the liver enzymes level including alanine transaminase, aspartate transaminase and alkaline phosphatase in serum and glutathione content in liver and kidney tissues. Other experiments such as Western blot analysis and quantitative real-time RT-PCR revealed that thymoquinone suppressed the expression of Bax/Bcl-2 ratio (both protein and mRNA level), and caspases activation pursuant to ethanol toxicity. CONCLUSION: This study indicates that thymoquinone may have preventive effects against ethanol toxicity in the liver and kidney tissue through reduction in lipid peroxidation and inflammation, and also interrupting apoptosis.

Effect of aqueous and ethanolic extracts of Pimpinella anisum L. seeds on milk production in rats.

Pimpinella anisum L. (P. anisum) is used as a galactagogue in traditional medicine; hence, the effect of aqueous and ethanolic extracts of P. anisum seeds on milk production in rats was evaluated. The milk production was assessed by measuring the pups' weights during the suckling period. The intraperitoneal LD(50) values of P. anisum aqueous and ethanolic extracts were 4.93 and 3.77 g/kg, respectively. The aqueous (1 g/kg) and ethanolic extracts (1 g/kg) increased the milk production significantly (p < 0.001), with about 68.1% and 81% more milk being produced, respectively, than in the control group. The pups gained weight during the study period with the aqueous (0.5 and 1 g/kg, p < 0.05) and ethanolic (0.5 and 1 g/kg, p < 0.01) extracts. Thus, P. anisum aqueous and ethanolic extracts can increase milk production in rats.

Effect of Nigella sativa fixed oil on ethanol toxicity in rats.

OBJECTIVES: This study was planned to appraise the protective effect of Nigella sativa fixed oil (NSO) against subchronic ethanol induced toxicity in rats. MATERIALS AND METHODS: Studies were carried out on six groups of six animals each, including control (normal saline, gavage), ethanol (3 g/kg/day, gavage), NSO (0.125, 0.25 and 0.5 ml/Kg/day, IP) plus ethanol and NSO (0.5 ml/Kg/day, IP) groups. Treatments were continued for 4 weeks. RESULTS: According to data, treatment with NSO attenuated ethanol-induced increased levels of malondialdehyde (MDA), tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6), as well as histopathological changes in liver and kidney tissues. Moreover, NSO improved the level of serum liver enzymes (including alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and glutathione (GSH) content in liver and kidney tissues in ethanol-treated rats. Western blot analysis and quantitative real time RT-PCR showed that NSO treatment inhibited apoptosis stimulated by ethanol through decreasing the Bax/Bcl-2 ratio (both protein and mRNA levels), cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 level in liver and kidney. CONCLUSION: This study showed that NSO may have protective effects against hepatotoxicity and renal toxicity of ethanol by decreasing lipid peroxidation and inflammation and preventing apoptosis.

Acute and subacute toxicity of safranal, a constituent of saffron, in mice and rats.

The acute and sub-acute toxicity of safranal were studied in rat and mice within 2 and 21 days after exposure, respectively. For subacute toxicity, changes in weight as well as biochemical, hematological and pathological parameters were studied. The intraperitoneal LD50 values of safranal were 1.48 mL/kg in male mice, 1.88 mL/kg in female mice and 1.50 mL/kg in male rats. Oral LD50 values were 21.42 mL/kg in male mice, 11.42 mL/kg in female mice and 5.53 mL/kg in male rats. For subacute toxicity, safranal was administered orally to male rats once daily for 21 days. In hematological tests, a significant decrease in RBC counts, hematocrit, hemoglobin and platelets were observed. Safranal decreased cholesterol, triglyceride and alkalin phosphatase. Lactate dehydrogenase and serum urea nitrogen were increased by safranal. Histological studies indicated that safranal did not have any toxic effect on the heart, liver and spleen. However, pathological changes were seen in the kidney and lung. According to LD50 values, safranal was low-toxic in acute intraperitoneal route and practically non-toxic in acute oral administration in both mice and rats. In subacute toxicity, safranal changed some hematological and biochemical parameters.

Anticonvulsant effect of Berberis integerrima L. root extracts in mice.

Berberis integerrima is a member of Berberidaceae family. Berberine is one of the main constituents of this plant, having neuroprotective effect on central nervous system diseases. In this study, the anticonvulsant activity of methanolic extract, and hydromethanolic fraction, and chloroform fraction of B integerrima was assessed. The anticonvulsant effect of B integerrima was investigated using both pentylenetetrazole (PTZ) and maximal electroshock (MES)-induced seizure models. The LD50 value of the methanolic extract was 302.676 mg/kg. In the PTZ test, methanolic extract (140 and 200 mg/kg, i.p., p<0.01), hydromethanolic fraction (200 mg/kg, p<0.01), and chloroform fraction (200 mg/kg, p<0.01) increased the onset time of hind limb tonic extensions (HLTEs). The protective effect against mortality (convulsion survivors/animals tested) was 2/8 in methanolic extract, and 3/8 in hydromethanolic fraction at a dose of 200 mg/kg and in chloroform fraction at a dose of 140 mg/kg. In the MES test, this plant did not display any significant effect in reducing HLTE duration. According to phytochemical screening, methanolic extract contained alkaloids and tannins. The present study, conducted in mice, indicated that B integerrima has anticonvulsant activity in PTZ-induced seizures. It is concluded that B integerrima may be useful in petit mal epilepsy.

Effect of aqueous and ethanolic extracts of Nigella sativa seeds on milk production in rats.

Nigella sativa L. is used as a galactagogue in traditional medicine. Hence, the effects of aqueous and ethanolic extracts of N. sativa seeds on milk production in rats were evaluated. The measurement of milk production was by measuring pup weight during suckling period. The intraperitoneal LD(50) values of aqueous and ethanolic extracts of N. sativa were 4.23 and 4.9 g/kg, respectively. The aqueous (0.5 g/kg) and ethanolic extracts (1 g/kg) increased milk production significantly (p < 0.001), producing about 31.3% and 37.6% more milk than control, respectively. During the study period, the pups gained weight with the aqueous (0.5 g/kg, p < 0.01) and ethanolic extracts (1 g/kg, p < 0.05). It is concluded that aqueous and ethanolic extracts of N. sativa can stimulate milk production in rats.

Protective Effect of Silymarin against Acrolein-Induced Cardiotoxicity in Mice.

Reactive α,ß-unsaturated aldehydes such as acrolein (ACR) are major components of environmental pollutants and have been implicated in the neurodegenerative and cardiac diseases. In this study, the protective effect of silymarin (SN) against cardiotoxicity induced by ACR in mice was evaluated. Studies were performed on seven groups of six animals each, including vehicle-control (normal saline + 0.5% w/v methylcellulose), ACR (7.5 mg/kg/day, gavage) for 3 weeks, SN (25, 50 and 100 mg/kg/day, i.p.) plus ACR, vitamin E (Vit E, 100 IU/kg, i.p.) plus ACR, and SN (100 mg/kg, i.p.) groups. Mice received SN 7 days before ACR and daily thereafter throughout the study. Pretreatment with SN attenuated ACR-induced increased levels of malondialdehyde (MDA), serum cardiac troponin I (cTnI), and creatine kinase-MB (CK-MB), as well as histopathological changes in cardiac tissues. Moreover, SN improved glutathione (GSH) content, superoxide dismutase (SOD), and catalase (CAT) activities in heart of ACR-treated mice. Western blot analysis showed that SN pretreatment inhibited apoptosis provoked by ACR through decreasing Bax/Bcl-2 ratio, cytosolic cytochrome c content, and cleaved caspase-3 level in heart. In conclusion, SN may have protective effects against cardiotoxicity of ACR by reducing lipid peroxidation, renewing the activities of antioxidant enzymes, and preventing apoptosis.

Protective effects of aqueous and ethanolic extracts of Portulaca oleracea L. aerial parts on H2O2-induced DNA damage in lymphocytes by comet assay.

The comet assay is a standard method for measuring DNA damage. In this study, the protective effects of ethanolic and aqueous extracts of Portulaca oleracea L. (P. oleracea) on human lymphocyte DNA lesions were evaluated with the comet assay. Lymphocytes were isolated from blood samples taken from healthy volunteers. Human lymphocytes were incubated in H(2)O(2) (50,100, and 200 µM), aqueous extract (0.05, 0.1, 0.5, 1, and 2.5mg/ml), and ethanolic extracts (0.05, 0.1, 0.5, 1, and 2.5mg/ml) of P. oleraceae aerial parts alone with a combination of H(2)O(2) (100 µM) with either 1 or 2.5mg/ml of both extracts at 4°C for 30 minutes. The extent of DNA migration was measured using the alkaline single cell gel electrophoresis approach assay, and DNA damage was expressed as percentage tail DNA. We found that the aqueous extract of P. oleracea significantly inhibited DNA damage, while there was no effect of the ethanolic extract. These data suggest that the aqueous extract of P. oleracea can prevent oxidative DNA damage to human lymphocytes, which is likely due to antioxidant constituents in the extract.