Protective Roles of Shilajit in Modulating Resistin, Adiponectin, and Cytokines in Rats with Non-alcoholic Fatty Liver Disease / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of Shilajit, a medicine of Ayurveda, on the serum changes in cytokines and adipokines caused by non-alcoholic fatty liver disease (NAFLD).@*METHODS@#After establishing fatty liver models by feeding a high-fat diet (HFD) for 12 weeks, 35 Wistar male rats were randomly divided into 5 groups, including control (standard diet), Veh (HFD + vehicle), high-dose Shilajit [H-Sh, HFD + 250 mg/(kg·d) Shilajit], low-dose Shilajit [L-Sh, HFD + 150 mg/(kg·d) Shilajit], and pioglitazone [HFD + 10 mg/(kg·d) pioglitazone] groups, 7 rats in each group. After 2-week of gavage administration, serum levels of glucose, insulin, interleukin 1beta (IL-1β), IL-6, IL-10, tumor necrosis factor-alpha (TNF-α), adiponectin, and resistin were measured, and insulin resistance index (HOMA-IR) was calculated.@*RESULTS@#After NAFLD induction, the serum level of IL-10 significantly increased and serum IL-1β, TNF-α levels significantly decreased by injection of both doses of Shilajit and pioglitazone (P<0.05). Increases in serum glucose level and homeostasis model of HOMA-IR were reduced by L-Sh and H-Sh treatment in NAFLD rats (P<0.05). Both doses of Shilajit increased adiponectin and decreased serum resistin levels (P<0.05).@*CONCLUSION@#The probable protective role of Shilajit in NAFLD model rats may be via modulating the serum levels of IL-1β, TNF-α, IL-10, adipokine and resistin, and reducing of HOMA-IR.

Aqueous date fruit efficiency as preventing traumatic brain deterioration and improving pathological parameters after traumatic brain injury in male rats

Objective: following traumatic brain injury, disruption of blood-brain-barrier and consequent brain edema are critical events which might lead to increasing intracranial pressure [ICP], and nerve damage. The current study assessed the effects of aqueous date fruit extract [ADFE] on the aforementioned parameters

Materials and Methods: in this experimental study, diffused traumatic brain injury [TBI] was generated in adult male rats using Marmarou's method. Experimental groups include two pre-treatment [oral ADFE, 4 and 8 mL/kg for 14 days], vehicle [distilled water, for 14 days] and sham groups. Brain edema and neuronal injury were measured 72 hours after TBI. Veterinary coma scale [VCS] and ICP were determined at -1, 4, 24, 48 and 72 hours after TBI. Differences among multiple groups were assessed using ANOVA. Turkey's test was employed for the ANOVA post-hoc analysis. The criterion of statistical significance was sign at P<0.05

Results: brain water content in ADFE-treated groups was decreased in comparison with the TBI+vehicle group. VCS at 24, 48 and 72 hours after TBI showed a significant increase in ADFE groups in comparison with the TBI+vehicle group. ICP at 24, 48 and 72 hours after TBI, was decreased in ADFE groups, compared to the TBI+vehicle. Brain edema, ICP and neuronal injury were also decreased in ADFE group, but VCS was increased following on TBI

Conclusion: ADFE pre-treatment demonstrated an efficient method for preventing traumatic brain deterioration and improving pathological parameters after TBI

effects of estrogen receptors' antagonist on brain edema, intracranial pressure and neurological outcomes after traumatic brain injury in rat

In previous studies, the neuroprotective effect of 17beta -estradiol in diffuse traumatic brain injury has been shown. This study used ICI 182,780, a non-selective estrogen receptor antagonist, to test the hypothesis that the neuroprotective effect of 17beta -estradiol in traumatic brain injury is mediated by the estrogen receptors. The ovariectomized rats were divided into eight groups. Brain injury was induced by Marmarou's method. Estrogen was injected 30 minutes after traumatic brain injury, and ICI 182,780 was injected before traumatic brain injury and also before estrogen treatment. In one group only ICI 182,780 was injected. The brain water content and Evans blue dye content were measured 24 and 5 hours after traumatic brain injury, respectively. The neurologic outcomes and intracranial pressure were assessed before, 4, and 24 hours after traumatic brain injury. Brain water content and Evans blue content were less in estrogen-treated group comparison to vehicle group. ICI 182,780 eliminated the effects of estrogen on brain edema and brain blood barrier permeability. Intracranial pressure was increased significantly after trauma, and estrogen decreased intracranial pressure at 4 and 24 hours after traumatic brain injury in comparison to vehicle. This inhibitory effect was also eliminated by treatment with ICI182,780. ICI 182,780 also inhibited the estrogen induced increase in neurologic outcomes following traumatic brain injury. However, the use of ICI 182,780 alone had no neuroprotective effect after traumatic brain injury. The results suggest that classical estrogen receptors have probably a role in the neuroprotective function of estrogen following traumatic brain injury

Neuroprotective antioxidant effect of sex steroid hormones in traumatic brain injury

The aim of the present study was to evaluate the effect of different doses of sex steroid hormones on brain edema, BBB permeability, brain antioxidant enzyme activity, and MDA level after traumatic brain injury [TBI] in ovarectomized [OVX] rats. Female rats were divided into six [One sham and 5 TBI] groups including: vehicle, estrogen in physiologic [33.3 microg/kg] and pharmacologic [1 mg/kg] doses, progesterone in physiologic [1.7 mg/kg] and pharmacological doses [8 mg/kg]. The results showed that compared to vehicle group, estrogen and progesterone groups showed significantly lower brain water content [P<0.001]. Evans blue content was significantly lower in both estrogen doses and in progesterone physiologic dose [P<0.001]. Evans blue content was significantly higher in progesterone pharmacologic dose [P<0.001]. Superoxide dismutase [SOD] activity was significantly higher in estrogen and progesterone pharmacologic doses [P<0.001]. Glutathione peroxidase [GPx] activity was significantly lower in estrogen physiologic dose [P<0.001]. It was concluded that the neuroprotective effect of different doses of sex steroid hormones after TBI, may be mediated by changes in oxidant agent activity

[Effects of different phases of estrous cycle on brain edema and neurological outcomes after severe traumatic brain injury in female rats]

Sex is one of the main recovery parameters after traumatic brain injury [TBI]. During estrus cycle phase, plasma estrogen and progesterone levels are very different. So in this study, we have assessed different effects of these phases in brain edema and neurological outcomes. Adult female Albino-N-MARI rats were divided into 6 groups [14 per group] including metestrus, diestrus, proestrus, estrus, ovarictomized [OVX] rats, and sham group [without TBI and ovary]. In all groups, brain water content for the measurement of brain edema, evans blue content for the measurement of brain vascular permeability, neurological scores, plasma estrogen and progesterone levels were assessed after severe TBI. Brain water content in diestrus, proestrus and estrus groups showed a significant decrease as compared with OVX group [P<0.001] but it was not significant in metestrus group. Evans blue content in proestrus group was lower than estrus and OVX groups [P<0.001]. Also, in diestrus group it was lower than OVX group [P<0.01].Neurological scores showed significant increases in proestrus group 4 hours after severe TBI as compared with metestrus, diestrus and OVX groups [P<0.01]. 24 hours after severe TBI, neurological scores in all groups were higher than OVX [P<0.001]. Brain water content and brain vascular permeability in sham group was lower than OVX, besides neurological scores in sham group were higher than OVX [P<0.001]. This study showed that brain edema, evans blue content and neurological scores after severe TBI are related to estrus cycle different phases. This could be related to differences in female sex hormones in different phases of estrus cycle

[ effect of sex steroid hormones on brain edema and intracranial pressure after experimental traumatic brain injury in rats]

We investigated the role of sex hormones on changes in brain edema intracranial pressure [ICP], cerebral perfusion pressure [CCP] after trauma brain injury [TBI] in ovarectomized female [OVX] rats. In this study female rats are divided into five groups. Control group [Intact] sham group and other groups include: vehicle, estrogen group [1mg/kg] and progesterone group [8 mg/kg] which on all groups TBI was induced by Marmarou method. 30 minutes after TBI, drugs were injected i.p. ICP was measured in spinal cord using a standard procedure. CPP was calculated by the mean arterial pressure [MAP] - ICP. Neurologic scores were measured by motor, eye and respiratory reflex. The results showed after TBI, water content was significantly lower in estrogen and progesterone groups [P<0.001] compared with vehicle group. Analysis showed a stable ICP up to 24 hours. The ICP in estrogen and progesterone groups was significantly decreased at 4 and 24 hours as compared to vehicle group [P<0.001in both cases]. The CPP at 24 hours after TBI, significantly increased in estrogen and progesterone groups compared with vehicle [P<0.001]. Also after TBI, neurologic scores was significantly higher in estrogen and progesterone groups as compared with vehicle [at 1 hours P<0.05, and at 24hours P<0.001 for estrogen], [at 1 hours P<0.01 for progesterone]. Our findings indicated an improvement of ICP, CPP and neurologic scores produced by pharmacologic doses of estrogen and progesterone after TBI in OVX rat. These effects may be contribute to neuroprotective effects of these hormones

Priming hepatitis B surface [HBsAg]- and core antigen [HBcAg]- specific immune responses by chimeric, HBcAg with a HBsAg 'a' determinant

We developed an immunogen to stimulate multivalent immunity against hepatitis B surface antigen [HBsAg] and hepatitis B core antigens [HBcAg]. Immune responses specific for both HBsAg and HBcAg play an important role in controlling the infection. HBsAg-specific antibodies mediate elimination of virions at an early stage of infection and prevent the spread of virus. The immunogen was constructed by inserting the immunodominant, antibody-binding 'a' determinant [aa 111-149] of HBsAg [with or without a poly-glycine [PG] linker] into the e2 epitope of HBcAg. Only the constructs in which the HBsAg 'a' determinant was inserted into HBcAg, flanked by PG linkers, expressed a chimeric protein in human embryonic kidney cells with HBsAg and HBcAg antigenicity. Both glycosylated and non-glycosylated forms of the chimeric protein were immunoprecipitated from cell lysate. Intramuscular DNA vaccination of mice with plasmids expressing chimeric HBcAg primed antibody responses against well-defined serologically-defined determinants of both, native HBcAg, and native HBsAg. In addition, CD8[+] T cell responses against HBcAg epitopes were primed by this chimeric HBV antigen. The e2 sequence of HBcAg can thus be used to present heterologous epitopes without loss of immunogenicity of the HBcAg protein

Expression of a Chimeric protein containing the catalytic domain of Shiga-like toxin and Human Granulocyte macrophage colony-stimulating Factor [hGM-CSF] in Escherichia coli and Its recognition by reciprocal antibodies

Fusion of two genes at DNA level produces a single protein, known as a chimeric protein. Immunotoxins are chimeric proteins composed of specific cell targeting and cell killing moieties. Bacterial or plant toxins are commonly used as the killing moieties of the chimeric immunotoxins. In this investigation, the catalytic domain of Shiga-like toxin [A1] was fused to human granulocyte macrophage colony stimulating factor [hGM-CSF] gene and the fused gene was then expressed using an expression vector containing arabinose promoter. The protein thus obtained could be recognized by two different ELISA system designed for detection of hGM-CSF and Shiga-toxin and reconfirmed by Western-blot. The recognition of the chimeric protein by specific antibodies could be indicative of the proper form of the protein, which justifies further steps to be taken to evaluate the potential effects of the chimeric protein

Construction of hybrid gene of hepatitis B surface antigen carrying heat-stable enterotoxin of escherichia coli and its expression in mammalian cell line

Hepatitis B surface antigen is the first genetically engineered vaccine licensed for human use. Various strategies have been proposed to obtain a vaccine that would bypass the need for injection. In this study, a non-toxic portion of heat-stable enterotoxin of Escherichia coli that is capable of adhering to epithelial cells was inserted at amino acid position 112 of hepatitis surface antigen. The construct was used for transfection of human embryonic kidney cells in order to assess the expression of the hybrid protein. The data obtained showed a very low level of expression. In vivo antibody production and cytotoxic T lymphocyte response in B6 mice were assessed using DNA immunization. Three out of five injected mice responded with titers 10 mIU/ml anti-HBsAg and cytotoxic T-lymphocyte response was much higher with construct encoding the chimeric protein. Although this study proves that the chimeric protein is capable of eliciting both humoral and cellular responses, but further work is required to fully explore the feasibility of combining the properties of the two proteins