Acetylcholinesterase Activity In Lymphocytes Research Articles

Multitarget Effect of 2-(4-(Methylthio)phenyl)-3-(3-(piperidin-1-yl)propyl)thiazolidin-4-one in a Scopolamine-Induced Amnesic Rat Model.

Cholinergic system dysfunction, oxidative damage, and alterations in ion pump activity have been associated with memory loss and cognitive deficits in Alzheimer's disease. 1,3-thiazolidin-4-ones have emerged as a class of compounds with potential therapeutic effects due to their potent anticholinesterase activity. Accordingly, this study investigated the effect of the 2-(4-(methylthio)phenyl)-3-(3-(piperidin-1-yl)propyl)thiazolidin-4-one (DS12) compound on memory, cholinergic and oxidative stress parameters, ion pump activity, and serum biochemical markers in a scopolamine-induced memory deficit model. Male Wistar rats were divided into four groups: I-Control; II-Scopolamine; III-DS12 (5mg/kg) + scopolamine; and IV-DS12 (10mg/kg) + scopolamine. The animals from groups III and IV received DS12 diluted in canola oil and administered for 7days by gavage. On the last day of treatment, scopolamine (1mg/kg) was administered intraperitoneally (i.p.) 30min after training in an inhibitory avoidance apparatus. Twenty-four hours after scopolamine administration, the animals were subjected to an inhibitory avoidance test and were thereafter euthanized. Scopolamine induced memory deficits, increased acetylcholinesterase activity and oxidative damage, and decreased Na+/K+-ATPase activity in cerebral cortex and hippocampus. Pretreatment with DS12 prevented these brain alterations. Scopolamine also induced an increase in acetylcholinesterase activity in lymphocytes and whereas butyrylcholinesterase in serum and treatment with DS12 prevented these changes. In animals treated with DS12, no changes were observed in renal and hepatic parameters when compared to the control group. In conclusion, DS12 emerged as an important multitarget compound capable of preventing neurochemical changes associated with memory deficits.

Caffeine and high intensity exercise: Impact on purinergic and cholinergic signalling in lymphocytes and on cytokine levels.

This study evaluated the effects of caffeine in combination with high-intensity interval training (HIIT) on sensitivity to glucocorticoids and proliferation of lymphocytes, IL-6 and IL-10 levels and NTPDase, adenosine deaminase (ADA) and acetylcholinesterase (AChE) activity in rat lymphocytes. The animals were divided into groups: control, caffeine 4 mg/kg, caffeine 8 mg/kg, HIIT, HIIT plus caffeine 4 mg/kg and HIIT plus caffeine 8 mg/kg. The rats were trained three times a week for 6 weeks for a total workload 23% of body weight at the end of the experiment. Caffeine was administered orally 30 min before the training session. When lymphocytes were stimulated with phytohaemagglutinin no changes were observed in proliferative response between trained and sedentary animals; however, when caffeine was associated with HIIT an increase in T lymphocyte proliferation and in the sensitivity of lymphocytes to glucocorticoids occurred. ATP and ADP hydrolysis was decreased in the lymphocytes of the animals only trained and caffeine treatment prevented alterations in ATP hydrolysis. HIIT caused an increase in the ADA and AChE activity in lymphocytes and this effect was more pronounced in rats trained and supplemented with caffeine. The level of IL-6 was increased while the level of IL-10 was decreased in trained animals (HIIT) and caffeine was capable of preventing this exercise effect. Our findings suggest that caffeine ingestion attenuates, as least in part, the immune and inflammatory alterations following a prolonged HIIT protocol.

Quercetin treatment regulates the Na+,K+-ATPase activity, peripheral cholinergic enzymes, and oxidative stress in a rat model of demyelination

Quercetin is reported to exert a plethora of health benefits through many different mechanisms of action. This versatility and presence in the human diet has attracted the attention of the scientific community, resulting in a huge output of in vitro and in vivo (preclinical) studies. Therefore, we hypothesized that quercetin can protect Na+,K+-ATPase activity in the central nervous system, reestablish the peripheral cholinesterases activities, and reduce oxidative stress during demyelination events in rats. In line with this expectation, our study aims to find out how quercetin acts on the Na+,K+-ATPase activity in the central nervous system, peripheral cholinesterases, and stress oxidative markers in an experimental model of demyelinating disease. Wistar rats were divided into 4 groups: vehicle, quercetin, ethidium bromide (EB), and EB plus quercetin groups. The animals were treated once a day with vehicle (ethanol 20%) or quercetin 50 mg/kg for 7 (demyelination phase, by gavage) or 21 days (remyelination phase) after EB (0.1%, 10 μL) injection (intrapontine).The encephalon was removed, and the pons, hypothalamus, cerebral cortex, hippocampus, striatum, and cerebellum were dissected to verify the Na+,K+-ATPase activity. Our results showed that quercetin protected against reduction in Na+,K+-ATPase in the pons and cerebellum in the demyelination phase, and it increased the activity of this enzyme in the remyelination phase. During the demyelination, quercetin promoted the increase in acetylcholinesterase activity in whole blood and lymphocytes induced by EB, and it reduced the increase in acetylcholinesterase activity in lymphocytes in the remyelination phase. On day 7, EB increased the superoxide dismutase and decreased catalase activities, as well as increased the thiobarbituric acid–reactive substance levels. Taken together, these results indicated that quercetin regulates the Na+,K+-ATPase activity, affects the alterations of redox state, and participates in the reestablishment of peripheral cholinergic activity during demyelinating and remyelination events.

Chagas disease: modulation of the inflammatory response by acetylcholinesterase in hematological cells and brain tissue.

Chagas disease is an acute or chronic illness that causes severe inflammatory response, and consequently, it may activate the inflammatory cholinergic pathway, which is regulated by cholinesterases, including the acetylcholinesterase. This enzyme is responsible for the regulation of acetylcholine levels, an anti-inflammatory molecule linked to the inflammatory response during parasitic diseases. Thus, the aim of this study was to investigate whether Trypanosoma cruzi infection can alter the activity of acetylcholinesterase and acetylcholine levels in mice, and whether these alterations are linked to the inflammatory cholinergic signaling pathway. Twenty-four mice were divided into two groups: uninfected (control group, n=12) and infected by T. cruzi, Y strain (n=12). The animals developed acute disease with a peak of parasitemia on day 7 post-infection (PI). Blood, lymphocytes, and brain were analyzed on days 6 and 12 post-infection. In the brain, acetylcholine and nitric oxide levels, myeloperoxidase activity, and histopathology were analyzed. In total blood and brain, acetylcholinesterase activity decreased at both times. On the other hand, acetylcholinesterase activity in lymphocytes increased on day 6 PI compared with the control group. Infection by T. cruzi increased acetylcholine and nitric oxide levels and histopathological damage in the brain of mice associated to increased myeloperoxidase activity. Therefore, an intense inflammatory response in mice with acute Chagas disease in the central nervous system caused an anti-inflammatory response by the activation of the cholinergic inflammatory pathway.

Cholinergic enzymes and inflammatory markers in rats infected by Sporothrix schenckii

The aim of this study was to evaluate the cholinesterase activity in serum, whole blood, and lymphocytes, as well as to verify its relation to immune response in rats experimentally infected by Sporothrix schenckii. For this study, 63 Wistar rats (Rattus norvegicus), male, adult were divided into three groups: the negative control group (GC: n = 21), the group infected subcutaneously (GSC: n = 21), and the group infected intraperitoneally (GIP: n = 21). The groups were divided into subgroups and the following variables were evaluated at 15, 30, and 40 days post-infection (PI): acetylcholinesterase (AChE) activity in lymphocytes and whole blood, butyrylcholinesterase (BChE) activity in serum, cytokines levels (IL-1, IL-6, TNFα, and INF-γ), immunoglobulins levels (IgA, IgG, IgM, and IgE), and protein profile by electrophoresis. Both infected groups showed increased levels of inflammatory parameters (P < 0.05) in tissue and inflammatory infiltrates. The activities of AChE in lymphocytes and BChE in serum increased (P < 0.05) significantly in animals from the GSC group on day 40 PI compared to the GC group. Regarding the GIP, there was a marked increase in the AChE activity in lymphocytes on days 30 and 40 PI, and in whole blood on days 15, 30, and 40 PI compared to GC. Furthermore, IL-10, an anti-inflammatory cytokine, was also present in high levels during chronic systemic S. schenckii infections in animals. Therefore, it is concluded that cholinesterase has an important modulatory role in the immune response during granulomatous infection by S. schenckii.

Individual in vitro effects of ochratoxin A, deoxynivalenol and zearalenone on oxidative stress and acetylcholinesterase in lymphocytes of broiler chickens.

The contamination of consumer food and animal feed with toxigenic fungi has resulted in economic losses worldwide in animal industries. Mycotoxins are highly biologically reactive secondary metabolites and can inhibit protein synthesis and cell multiplication. Considering the cytotoxicity of mycotoxins, this experiment was performed to determine the in vitro influence of ochratoxin A, deoxynivalenol and zearalenone on lipid peroxidation in lymphocytes of broiler chickens at different concentrations. This study has also evaluated whether the presence of these mycotoxins changes the acetylcholinesterase activity in lymphocytes, which is involved in the regulation of immune and inflammatory responses. Blood lymphocytes of broiler chickens were isolated through density gradient centrifugation and incubated with the respective mycotoxins at concentrations of 0.001, 0.01, 0.1 and 1 μg/mL. Lipid peroxidation, which was evaluated through the amount of malondialdehyde measured in a thiobarbituric acid-reactive species test, and the enzymatic activity were analyzed at 24, 48 and 72 h. Results of the lipid peroxidation evaluation showed an increasing cytotoxicity relation: ochratoxin A > deoxynivalenol > zearalenone. Conversely, cytotoxicity was valued as zearalenone > deoxynivalenol > ochratoxin A in relation to the acetylcholinesterase enzymatic activity. At a concentration of 1 μg/mL, ochratoxin A and deoxynivalenol induced the highest cellular oxidative stress levels and the highest enzymatic activity at the majority of time points. However, the same mycotoxins, except at 1 μg/mL concentration, induced a reduction of lymphocytic lipid peroxidation 72 h after incubation, suggesting the action of a compensatory mechanism in these cells.

Alterations of ectonucleotidases and acetylcholinesterase activities in lymphocytes of Down syndrome subjects: Relation with inflammatory parameters

BackgroundSubjects with Down syndrome (DS) have an increased susceptibility to infections and autoimmune disorders. ATP, adenosine, and acetylcholine contribute to the immune response regulation, and NTPDase, adenosine deaminase (ADA) and acetylcholinesterase (AChE) are important enzymes in the control of the extracellular levels of these molecules. We evaluated the activities of these enzymes and the cytokine levels in samples of DS individuals. MethodsThe population consisted of 23 subjects with DS and 23 healthy subjects. Twelve milliliters of blood was obtained from each subject and used for lymphocyte and serum preparation. Lymphocytes were separated on Ficoll density gradients. After isolation, NTPDase and AChE activities were determined. ResultsThe NTPDase activity using ADP as substrate was increased in lymphocytes of DS patients compared to control (P<0.05); however, no alterations were observed in the ATP hydrolysis. An increase was observed in the AChE activity in lymphocytes and in ADA activity in serum of DS patients when compared to healthy subjects (P<0.05). In DS subjects, an increase in the levels of IL-1β, IL-6, TNF-α and IFN-γ and a decrease in the IL-10 levels were also observed (P<0.05). ConclusionsAlterations in the NTPDase, ADA and AChE activities as well changes in the cytokine levels may contribute to immunological alterations observed in DS.

Influence of experimental infection by Haemonchus contortus on acetylcholinesterase activity in lymphocytes of lambs

The aim of this study was to evaluate the acetylcholinesterase (AChE) activity in lymphocytes of lambs experimentally infected by Haemonchus contortus. A total of 14 healthy lambs were used, divided into two groups of seven animals each. Group A (negative control) represented the uninfected animals, and Group B (positive control) was formed by animals infected with 15,000 larvae of H. contortus. Blood was drawn on the days 15, 45 and 75 post-infection (PI) in order to perform the white blood cells (WBC) count, as well as the evaluation of AChE activity in lymphocytes. Parasitological stool exam (eggs per gram of feces – EPG) was performed on the same days to follow up the evolution of the infection. On day 15 PI it was verified negative EPG; however, on days 45 and 75 PI it was observed positive EPG only in the animals of group B. In the three evaluated periods was observed a lower number of leukocytes, associated with decreased lymphocytes and neutrophils in lambs infected by this gastrointestinal nematodes. Lambs infected with H. contortus showed significant (P<0.01) lower AChE activity in lymphocytes compared uninfected. Statistically, there was a positive correlation (P<0.05) between AChE activity in lymphocytes and number of lymphocytes (r=0.69). The lymphocytes are cells with direct participation in the cholinergic system; therefore, based on these results, it can be concluded that the experimental infection with H. contortus influences the number of lymphocytes, and consequently the AChE activity in these cells.

Complete blood count and acetylcholinesterase activity of lymphocytes of demyelinated and ovariectomized rats treated with resveratrol

Resveratrol is a phytoestrogen that has many beneficial actions. This study aimed to evaluate the effect of resveratrol on the complete blood count (CBC) and the acetylcholinesterase (AChE) activity of lymphocytes of ovariectomized rats experimentally demyelinated by ethidium bromide (EB). Forty adult female Wistar rats (60 days, 200–220 g) were divided randomly into five groups (n = 4) to evaluate the demyelination phase and five groups (n = 4) to evaluate the remyelination phase. In each phase, the groups consisted of sham rats–G1; ovariectomized rats, not demyelinated, treated only with vehicle (ethanol 25%)–G2; demyelinated ovariectomized rats treated only with vehicle–G3; ovariectomized rats, not demyelinated, treated with resveratrol–G4; and demyelinated ovariectomized rats treated with resveratrol–G5. Only during the remyelination phase, CBC showed a significant difference (p < 0.05) in the number of monocytes between G2 and G5 groups. In the demyelination phase, there was a significant decrease (p < 0.05) in the AChE activity in the G4 group, while the G5 group was statistically similar to the G1, G2 and G4 groups. In the remyelination phase, there were no significant differences in the AChE activity among the groups. The treatment for 7 days with resveratrol with or without the experimental demyelization with EB appears to influence the AChE activity of lymphocytes, without changing the number of these cells in the circulation. However, in the remyelination phase, there seems to be stabilization in its effect on the lymphocyte AChE activity.

Cholinesterases as markers of the inflammatory process in rats infected with Leptospira interrogans serovar Icterohaemorrhagiae

The aim of this study was evaluate changes in the cholinesterase activity in blood, lymphocytes and serum of rats infected with Leptospira interrogans serovar Icterohaemorrhagiae ('L. icterohaemorrhagiae'). Sixty adult Wistar rats were divided into six groups of 10 animals: three control groups and three test groups. The animals from the test groups were intraperitoneally inoculated with 1 ml medium containing 1 × 10(8) leptospires. The activity of acetylcholinesterase in blood and butyrylcholinesterase in serum increased on days 5 (P<0.05) and 30 (P<0.021) post-infection, respectively. A decrease in lymphocyte count was observed on days 15 (P<0.01) and 30 post-infection (P<0.05). On day 15 post-infection, acetylcholinesterase activity (P<0.001) in lymphocytes decreased in infected rats. However, on day 30 post-infection there was an increase in acetylcholinesterase activity in lymphocytes. In conclusion, our results showed that the activity of enzymes of the cholinergic system in the total blood, lymphocytes and serum is altered as a result of inflammation caused by infection with L. icterohaemorrhagiae. The possible causes of these alterations will be discussed in this paper.

Trypanosoma evansi: Immune response and acetylcholinesterase activity in lymphocytes from infected rats

The existence of cholinergic receptors in the immune system cells is well documented. This study aimed to evaluate the acetylcholinesterase activity (AChE) in lymphocytes from rats infected with Trypanosoma evansi in acute and chronic phase disease. Twenty animals were infected with 106 trypomastigotes forms each and 10 were used as negative controls. The two groups of inoculated rats were formed according to the degree of parasitemia and the period post-infection (PI). Group A: rats with 4days PI and between 24 and 45parasites/field (1000×); group B: rats with 30days PI and parasitemia with jagged peaks between 0 and 1parasites/field; group C: not-infected animals. At 4days PI (acute phase) and 30days PI (chronic phase) the rats were anesthetized to collect blood for hemogram and separation of lymphocytes. After separation, the AChE activity was measured in lymphocytes. It was observed that the number of lymphocytes increased significantly in group A compared to group C. The activity of AChE in lymphocytes significantly increased in acute phase and decreased in chronic phase in the infected rats when compared to not-infected (P<0.05). Statistical analysis showed a positive correlation between the number of lymphocytes and AChE activity in lymphocytes in 4days PI (r2: 0.59). Therefore, the infection by T. evansi influences AChE activity in lymphocytes of rats indicating changes in the responses of cholinergic system in acute phase, possibly due to immune functions performed by these enzymes.

Blood Cells Cholinesterase Activity in Early Stage Alzheimer’s Disease and Vascular Dementia

Blood acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities have been studied as markers for Alzheimer’s disease (AD), but their usefulness as a disease marker is controversial. To determine cholinesterase (ChE) activity during AD progression and whether ChE changes associate to other dementias, ChE activity was measured in lymphocytes, erythrocytes and platelets. Subjects underwent extensive medical and neuropsychological examination. Both early-AD and AD patients had lower AChE activity in lymphocytes compared to control subjects (p < 0.0001). In contrast, erythrocyte AChE activity was higher in patients with vascular dementia (p = 0.004). Low ChE activity in lymphocytes was the best discriminator for AD. Because it was already low at very early stages of AD, ChE could be helpful as an early biomarker of differential diagnosis for the follow-up of patients during their early stages of cognitive impairment before a clinical dementia is established.

Modulation of Muscarinic Receptors and Acetylcholinesterase Activity in Lymphocytes and in Brain Areas Following Repeated Organophosphate Exposure in Rats

Modulation of Muscarinic Receptors and Acetylcholinesterase Activity in Lymphocytes and in Brain Arcas Following Repealed Organophosphate Exposure in Rats. Fitzgerald, B. B., and Costa, L. G. (1993). Fundam. Appl. Toxicol. 20, 210-216.Repeated exposures to organophosphorus (OP) insecticides has been shown to cause a decrease of cholinergic muscarinic receptors (mAChR) in brain and in peripheral tissues. These changes are believed to be involved in the development of tolerance to OP toxicity and may play a role in cognitive dysfunctions observed following repeated OP exposure. Recently, mAChRs identified in circulating lymphocytes have been shown to be modulated similarly to brain mAChRs following repealed OP exposure, suggesting that these peripheral cells may be useful as indicators of mAChR changes in the central nervous system. This study was designed to further investigate whether mAChRs on lymphocytes could serve as a biomarker for changes in brain mAChRs during prolonged OP exposure and during recovery from such exposure. Using the mAChR antagonist [3H]quinuclidinyl benzilate (QNB) to label mAChRs, we found that exposure to disulfoton for 14 days (2 mg/kg/day by gavage) caused a significant decrease (25-35%) in muscarinic receptors density in the cerebral cortex, hippocampas, and striatum, as well as in circulating lymphocytes. The decline of mAChR density in lymphocytes paralleled those observed in brain, particularly in cortex and hippocampus, during exposure to disulfoton; however, while brain mAChR levels recovered slowly after termination of exposure and remained significantly reduced 4 weeks after the last treatment, [3H]QNB binding in lymphocytes recovered rapidly within 1 week. Similarly, lymphocyte acetylcholinesterase (ACHE) activity was significantly inhibited and correlated well with brain AChE activity during exposure, but the recovery was rapid relative to AChE activity in brain. On the other hand, the recovery of red blood cell AChE paralleled that observed in brain tissue. These findings suggest that lymphocytes may be useful as peripheral markers to monitor for changes in mAChR and AChE in brain tissue during repeated OP exposure, but are not suitable markers for these CNS parameters after termination of exposure.

Modulation of Muscarinic Receptors and Acetylcholinesterase Activity in Lymphocytes and in Brain Areas Following Repeated Organophosphate Exposure in Rats

Modulation of Muscarinic Receptors and Acetylcholinesterase Activity in Lymphocytes and in Brain Areas Following Repeated Organophosphate Exposure in Rats

Changes in the cholinergic system of lymphocytes due to mitogenic stimulation

A significant increase in acetylcholinesterase (AChE) activity and muscarinic-type cholinergic receptor (mAChR) expression was demonstrated in normal human peripheral blood lymphocytes as an early response to various stimuli. AChE activity of lymphocytes from T-cell acute lymphoid leukemia patients, and that of leukemic cell lines was enhanced or increased only slightly, in good accordance with their significantly decreased response to mitogenic stimuli. The higher initial AChE activity of cultured leukemic cell lines probably reflects their permanently activated state. AChE activity and mAChR expression in these cases could only be increased by a differentiation-inducing agent. Based on these observations, the possible role of the cholinergic modulation in thymic lymphocyte maturation is assumed.

Altered lymphocyte acetylcholinesterase activity in patients with senile dementia

Reductions in the acetylcholinesterase (AChE) activity of certain brain areas in patients with senile dementia of Alzheimer type (SDAT) have been found to correlate with the severity of the disease, suggesting a central cholinergic lesion. Since AChE is expressed on the surface of various blood cells too, the AChE activity of lymphocytes and erythrocytes was determined to test the possibility whether the cholinergic lesion is also reflected on these readily available cells. The AChE activity of lymphocytes in SDAT and in alcoholic dementia (AD) were significantly lower as compared to those of the age-matched healthy volunteers. In contrast, there was no significant difference in the activity of lymphocyte AChE between age-matched healthy controls and patients with multi-infarct dementia of vascular origin (MID). No changes could be demonstrated in the erythrocyte AChE activities of the patients studied, and the age-matched healthy individuals, when comparing them to the healthy blood donors. The AChE activity of lymphocytes may thus be a useful marker to follow the alterations in the metabolism of acetylcholine (ACh) in the central nervous system (CNS) of different types of dementia.

Acetylcholinesterase activity of lymphocytes: an enzyme characteristic of T-cells.

Acetylcholinesterase (AchE) activity of normal human peripheral blood lymphocytes was determined by an adaptation of the colorimetric and radiometric techniques described for other cells. The enzyme activity seemed to be correlated to the number of T-lymphocytes in the blood. To decide whether only the T-cells possess AchE activity the lymphocytes were separated on Leucopac filter and on Percoll density gradient. B-lymphocytes had no detectable enzyme activity, while the T-lymphocyte fraction represented the total activity measured in the unseparated sample. The majority of AchE activity could be demonstrated in T-lymphocytes of lower density (TLD). The role of AchE in the plasma membrane of various blood cells is not known. Nevertheless, the enzyme is a good marker of the integrity and functional state of the membrane. The difference observed in AchE activity of the lymphocyte populations seems to be suitable for using it to characterize T-lymphocytes.