Additionally the influence of different benzodiazepines on the mitogen response of human peripheral blood lymphocytes and spleen cells was tested. Diazepam led to earlier disease onset, prolonged duration of symptoms, higher weight loss and overall disease index in VACV infected mice. For this reason, we decided to treat diseased mice therapeutically with DZ when they reached the clinical score of 1. Most importantly, this treatment ameliorated clinical signs, reduced the MOG-specific inflammatory cytokine production and prevented axonal damage.
Altogether, these results indicate that DZ is a potent immunomodulator capable of controlling undesired innate and adaptive immune responses, both at the beginning of these responses and also once they have started. Benzodiazepines are psychoactive drugs that share similar pharmacological properties, such as sedative, hypnotic sleep-inducing , anxiolytic, and anticonvulsive action and are widely used adjuncts to anesthesia to induce central muscle relaxation and amnesia 1.
In addition, these drugs also affect immunity. The effect of benzodiazepines depends on the activation of binding sites, such as the central and peripheral benzodiazepine receptors.
The peripheral benzodiazepine receptor is a ubiquitously expressed protein of the outer mitochondrial membrane termed translocator protein 18 kDa TSPO , structurally and functionally different from the GABA A receptor 3. The TSPO is expressed in platelets, immune cells, endothelium, vascular smooth muscle, bone marrow, endocrine cells and to a lesser extent in the CNS where it is associated with glial cells 4 , 5.
Upregulation of the TSPO is observed in many CNS diseases, and some TSPO ligands are currently under investigation as therapeutic means for promoting neuroprotection, axonal regeneration, and modulating inflammation 6 , 7.
Diazepam DZ , which is a mixed-type benzodiazepine that can act on both central and peripheral benzodiazepine receptors, has been demonstrated to have an inhibitory effect on T cell function 8 — In this work, we studied whether DZ impairs two undesired immune responses as septic shock and murine experimental autoimmune encephalomyelitis EAE. The invasion of microbial pathogens into the bloodstream is characterized by a systemic pro-inflammatory response, which can lead to severe sepsis and septic shock.
Here we described the bias from the classical activation of innate cells induced by LPS towards anti-inflammatory profiles when cultured with DZ, which prevented acute responses dependent on these cell populations.
DCs are professional antigen-presenting cells and have a key role in initiating and regulating adaptive immune responses. Both immature and semi-mature DCs have been associated with an induction of tolerance through the generation of T reg cells, the induction of apoptosis, or the anergy of autoreactive effector cells 15 — In this work, we demonstrate the impaired LPS-induced activation when DC were co-treated with DZ and its inability to initiate Th1 and Th17 adaptive inflammatory responses.
On the other hand, EAE is a well-accepted model that mimics many of the clinical and pathological features of multiple sclerosis MS. This model is characterized by a high induction of Th1 and Th17 autoimmune responses, mononuclear inflammatory infiltration and demyelination. Here, we show that DZ not only prevented the onset of autoimmune responses, but more importantly, it also improved the clinical signs when it was administered therapeutically in vivo , once the disease was established, maintaining a low clinical score until 35 days post-immunization dpi.
In summary, this work describes the immunomodulatory effects of Diazepam on the different stages of the immune response. DZ interferes with the activation of innate cells such as dendritic cells and macrophages induced by inflammatory stimuli and impairs the initiation and development of adaptive inflammatory responses Th1 and Th Furthermore, DZ also favors the development of tolerogenic and anti-inflammatory responses such as Tregs or antigen-specific IL producing cells.
Our work contributes in a descriptive way to the knowledge of the immunomodulatory properties of this type of psychoactive drugs. The cells were cultured in a concentration 1.
The concentration of nitrites was analyzed by a standard Griess reaction adapted to microplate as an indirect measurement of nitric oxide synthesis. The data were referred to a standard curve of sodium nitrite After 12 h of the last DZ injection, each mouse was i.
Louis, Mo, USA. The onset of systemic clinical signs, including reduced mobility, lethargy, shivering, piloerection, and congested conjunctiva was recorded.
Survival rate was determined every 6 h for 3 days post-septic shock induction, and then mice were euthanized by carbon dioxide overdose. DCs were generated as previously described The direct effect of DZ on cell viability, activation status, production of cytokines, and activation of intracellular signaling pathways mediated by LPS was evaluated.
In general, DCs differentiated from bone marrow were cultured in a concentration 1. Cell viability was assessed with 3- 4,5-dimethylthiazolyl -2,5-diphenyltetrazolium bromide MTT.
Absorbance was recorded at nm on a microplate reader After 15 days, the animals were sacrificed and splenocytes collected. Data were analyzed using Flowing software 2. Control mice were immunized with CFA followed by pertussis toxin. Mice were weighed and clinically scored daily. The onset of the disease correlates with weight loss, which might begin 1—2 days before EAE signs are visible and can be used as an indicator of disease activity.
Clinical signs usually begin between days 9 and 14 and were scored as follows: 0, no deficit; 0. The mean clinical score was defined as the sum of the highest clinical score achieved by each mouse during the acute phase of the disease divided by the number of mice At 30 or 35 days dpi the animals were euthanized. Then, 1. The axons in the medial dorsal column on the spinal cord at C5 were manually quantified.
The dorsal columns are reproducibly damaged in this model of EAE though the rostral—caudal level of damage was less predictable , and C5 was chosen to include ascending fibers from both the upper and lower limbs in the count Using Fiji software an inverted triangular ROI was drawn within the gracile fasciculus: a line from the spinal canal to the dorsal surface was bisected to indicate the dorsal half of the column height of the triangle and a line perpendicular to this was drawn on both sides from the most medial part of the column to the ends of gracile fasciculus on the dorsal surface as described.
Axon count was performed using cell counter plugin for Fiji. The murine microglial cell line BV2 was a kind gift from Dr. Motoneuron-derived 1 MN-1 cells, which are hybrids derived from embryonic mouse spinal cord motor neurons, were a kind gift from Dr. As the largest medical workforce, GPs have the greatest contact with patients and therefore prescribe the most medications. They are often seen as responsible for the benzodiazepine problem. But GPs may not always be the initiator. They often take over the care of patients who have been started on benzodiazepines by psychiatrists or during hospital admissions.
Psychiatrists commonly prescribe these medications as an add-on to antidepressant treatment to reduce anxiety and to increase the likelihood that patients will adhere to treatments and respond early. Today there is a broad spectrum of people and contexts where benzodiazepines are used, from the clearly legitimate short-term situations in known patients to deliberate drug misuse and sale in people with chaotic and sometimes aggressive behaviour. Further, with an increasingly internationally networked world, benzodiazepines can now be purchased on the internet without a prescription.
The promotion of the dangers of inadequate sleep also results in people seeking medications to reach the goal of seven to nine hours of sleep. In some cases, there are underlying causes that can be treated or healthy sleep habits can improve sleep.
In other cases, a person may feel well and simply need less sleep. New guidelines now recognise individual variations with a broader range of possible appropriate sleep durations of six to ten hours in adults aged years, and five to nine hours in those aged 65 years and over. Clinicians need to discuss the potential side effects of sedative drugs with patients, emphasising the risk of dependence and cognitive decline.
They also need to promote non-drug approaches for managing stress, insomnia and anxiety, such as cognitive behavioural therapy, and reinforce that sedative drugs do not work in the long term. Portsmouth Climate Festival — Portsmouth, Portsmouth. Dendritic cells DC are key to the beginning of antigen specific responses and the state of activation achieved by them defines the type of response that will be generated. We evaluated the effect of Dz on the activation of DC.
Dz did not affect the viability of resting or stimulated DC at any dose assayed. Keywords: Diazepam, dendritic cell, Macrophages, Lymphocytes, Experimental autoimmune encephalomyelitis. Presentation Type: Poster Presentation. Topic: Autoimmunity. Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.
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