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Cholestasis caused by the excessive accumulation of bile within the liver, due to intrinsic or extrinsic factors. Cholestasis side effects are associated directly or indirectly with the reduction of bile flow and the confinement of materials related to bile secretion (such as bile acids, bilirubin, and cholesterol). On the other hand, some factors such as opioids, alkaline phosphatase, endotoxin and nitric oxide increase in blood, which could cause tissue damage. Since water intake reduces during cholestasis and hypothalamic nuclei such as paraventricular and supraoptic nucleuses are involved in the regulation of body water; Therefore, in this study, the histopathological changes of hypothalamic nuclei were evaluated. Male Wistar rats weighing 200–250 g were randomly divided into three groups. Three sets of seven groups were unoperated control, sham-operated and bile duct-ligated rats. The tissue samples were analyzed using histotechnique and light microscope. Brain tissue necrosis in paraventricular and supraoptic nucleus in cholestatic rats increased, but in the sham and control rats no changes were observed and also cholestasis caused wrinkle chromatic nuclei and increased thickness of hypothalamic nuclei. Because endotoxin causes tissue trauma, it is likely increased endotoxin may leads to tissue changes in the brain.

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The aim of this study was to assess the effect of the smoke of Verbascum speciocum leaves grown in 'Bouein Zahra' region, on the healing of skin lesions in Wistar rats. In this study in vivo conditions, 36 adult male Wistar rats were randomly divided into 3 groups. The operation day was considered as day zero and the procedure took 10 days. After the end of the treatment, i.e., on the 14^th day, rats were killed by choloroform and samples were collected for histological analysis Masson trichrome and Vangason staining for collogen and hydroxyprolline amino acid OD assessment was applied. Studies in experimental groups exposed to the smoke of the leaves revealed that there was no increase in epidermis thickness, hair follicles diameter, skin, nor in the number and diameter of blood vessels as compared with control and sham groups. However, the amount of hair follicles was decreased. There was an increase in collogen diameter but this increase was not significant. But a significant increase was observed in OD of hydroxyl proline amino acid assessment (p<0.001) in experimental groups compared with sham and control groups. The results indicated that the use of smoke of V.speciocum leaves on the skin lesion had absolutely no increase on the wound healing ratios and its closure; it maximally could had very little effect on the proliferation as one step in the repair process, which needs further investigation.

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There are multiple neurotransmitters and neuromodulator systems mediating memory formation among which the endocannabinoid system plays a critical role in the memory formation by modulating the release of many neurotransmitters. Nucleus accumbens appears to have a site in the central of neuronal circuits of the limbic system and to be responsible for the integration and consolidation of inputs from other parts of the brain. In this study the influence of bilateral intra-nucleus accumbens shell microinjections of cannabinoid receptor agents on memory consolidation in adult male rats using passive avoidance task was investigated. The results showed that the intra-accumbens shell microinjection of ACPA as a CB1 receptor agonist (6 ng/rat) immediately after training decreased passive avoidance memory consolidation, while administration of its antagonist (AM251) at different doses did not affect passive avoidance memory consolidation. However, co-administration of AM251 (60 ng/rat) with an effective dose of ACPA prevented the impairment memory consolidation induced by ACPA. These results suggest that the accumbens shell cannabinoid system as a modulating system is involved in aversive memory consolidation including passive avoidance memory.

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Basolateral Amygdala is an important site of anxiety. Interactions between α-adrenergic and opioidergic systems in Basolateral amygdale were used for investigation anxiety and memory. The elevated plus-maze has been employed. The male wistar rats were used for this test. The site of BLA was cannulated bilaterally. Rats we injected morphine (4, 5.6 mg/kg) intraperitonealy, while clonidine (1, 2, 4 µg/rat) and yohimbin (0.5, 1,2 µg/rat) were injected to BLA. Open arm time percentage (%OAT), open arm entry (%0AE) and locomotor activity were determined by this behavioral test. Retention tested 24 hours later. Administration of morphine (6mg/kg) increased the OAT% in anxiety test, indicating anxiolytic-like effect. Intra Basolateral amygdala infusion of clonidine (4μg/rat) has an anxiolytic-like effect. While co-administration clonidine (4μg/rat) and in effective dose of morphine (4mg/kg) showed significant increase of OAT% in anxiety test; thus presenting anxiolytic response. Intra Basolateral amygdala administration of yohimbine (2μg/rat) decreased OAT% indicating of decrease anxiety-like behavior. While co-administration of intra Basolateral amygdala clonidine (4μg/rat) and effective dose of morphine (6mg/kg) showed a significant increase of OAT%, presenting anxiolytic response; co-administration of ineffective doses of morphine (4mg/kg) and yohimbine (1μg/rat) with the effective dose of clonidine (4μg/rat) showed that yohimbine could reverse the anxiolytic-like effect of morphine and clonidine. It should be noted that there are no significant changes in locomotor activity. The results indicate that morphine creates the compromise changes in adrenergic neurons of Basolateral amygdala by changing the α-adrenergic system on anxiety.                                                                                                                                                                  

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In cholestasis syndrome, the change of bile acids, salts, endotoxins and opioidsis is accompanied with hepaticencephalopathy and brain trauma; therefore, in this study, the histopathological changes of hippocampus after bile duct ligation were investigated. This study was performed on male Wistar rats. The rats were divided into three groups, namely the control, sham and cholestatic groups. After two weeks, the rats were killed under anesth-esia and their brains were dissected with no delay. The specimens were processed routinely and were sectioned into slices of 6-micron thickness. The sections were stained by Hematoxiline-Eosin (H&E) method. Subsequently, they were studied using optical microscope. Necrotic cells, the diminution of cell density and wrinkled chromatic nuclei were observed in hippocampus section. In addition, the thickness of hippocampus was mostly decreased. However, in the sham and control groups no changes were observed. Since endotoxin causes tissue trauma, the increase of en-dotoxin may result in tissue changes in the brain.

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Impaired motor functions were reported in cholestatic animals. This disorder in the function and death of motor neurons is highly dependent on changes in the environment around astrocytes and the blood-brain barrier, which is moderated by the aquaporin 4 protein. For this reason, the effects of cholestasis on motor cortex histology and morphology and aquaporin 4 protein levels were investigated in this study. Samples were stained by hematoxylin-eosin method. Histological changes in cortical brain were investigated. The amount of AQP4 protein in control, sham, and experimental groups were tested by immunohistochemistry. The thickness of motor cortex in cholestatic samples increased in comparison with the control and sham groups. Also, cholestasis caused wrinkle chromatic nuclei. On the other hand, tissue necrosis was detected in cholestatic group compared with sham and control groups. Reduction of cells densities in some cortical layers has been observed, which is probably indicative of cholestasis-induced cell death. AQP4 expression significantly decreased in BDL (p <0.05), but not in other groups (P<0.05). In this study, the pathology of motor cortex, which has also been associated with the decrease of neurons, could be considered the cause of motion abnormalities and AQP4 level reduction in cholestatic rats.