{"id":19546,"date":"2018-03-25T11:58:13","date_gmt":"2018-03-25T11:58:13","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=19546"},"modified":"2020-04-23T04:28:48","modified_gmt":"2020-04-23T04:28:48","slug":"bone-marrow-derived-protect-against-haloperidol-induced-brain-and-liver-damage-in-mice","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol11no1\/bone-marrow-derived-protect-against-haloperidol-induced-brain-and-liver-damage-in-mice\/","title":{"rendered":"Bone Marrow-Derived Protect Against Haloperidol-Induced Brain and Liver Damage in Mice"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Haloperidol is a classic antipsychotic drug in use to treat symptoms of schizophrenia. The drug acts by inhibiting dopamine D2 receptors (D2, 3, 4) in mesolimbic dopaminergic pathway1<\/sup> and is effective in improving the positive symptoms i.e., delusions and hallucinations.2<\/sup>\u00a0Haloperidol also binds to striatal D2 receptors with high occupancy and with an increased propensity of inducing extrapyramdial manifestations eg., tardive dyskinesia.3<\/sup>\u00a0The drug in addition is likely to impair cognition compared to the newer generation antipsychotic agents.4<\/sup>\u00a0Haloperidol causes brain oxidative stress,5-7<\/sup>\u00a0a situation where the increased generation of reactive oxygen metabolites overwhelms the cell\u2019s antioxidant mechanisms with a resultant oxidative damage to cell membrane lipids, enzymes, and nucleic acids.8<\/sup>\u00a0When given to rats, haloperidol resulted in increased lipid peroxidation, depletion of reduced glutathione, increased nitric oxide in several brain regions7,9,10<\/sup> and caused neuronal damage.11,12<\/sup>\u00a0Haloperidol thus despite its ability to improve symptoms of schizophrenia, might in the same time increase the susceptibility of the brain tissue to neurodegeneration via<\/em> causing oxidative stress. The latter has been linked to the development of schizophrenic symptoms13,14<\/sup> and there is evidence suggesting a benefit from administering antioxidant molecules to schizophrenic subjects.15-17<\/sup>\u00a0The presence of increased concentrations of the pro-inflammatory cytokines INF-g and NF-\u03b1 in the brain of schizophrenic subjects also indicates the presence of an ongoing inflammatory process18-20<\/sup> and this could be exacerbated by haloperidol.<\/p>\n

Stem cells are characterized by self-renewal and the ability to differentiate into a variety of cell lineages.21<\/sup>\u00a0They are classified into embryonic stem cells, embryonic germ cells and adult stem cells that can be found in bone marrow, blood, adipose tissue, brain, and liver.21,22<\/sup>\u00a0Research on stem cells has attracted much interest for their possible application in the treatment of many neurological diseases. Replacing diseased or damaged brain cells with other healthy cells is in itself an old hope and requirement.23<\/sup>\u00a0Recently, however, mounting evidence suggest that the benefit from stem cells might derive from an anti-inflammatory and immune-modulating actions.24<\/sup>\u00a0Mesenchymal stem cells (MSCs) can be isolated from adult tissues such as bone marrow, adipose tissue, muscle, liver, and skin.21\u00a0<\/sup>When given intravenously, these cells have demonstrated a therapeutic potential in experimental models of traumatic brain injury,24,25<\/sup>\u00a0Parkinson\u2019s disease,26<\/sup>\u00a0fulminant hepatic failure,27<\/sup>\u00a0liver fibrosis,28<\/sup>\u00a0graft-versus-host disease29<\/sup> and ischaemic acute renal failure.30<\/sup>\u00a0In these studies, stem cell therapy have shown to exert acute anti-inflammatory, antiapoptotic and immune modulating effects that are largely thought to account for the therapeutic benefit.<\/p>\n

In view of the above, the present study aimed to investigate the possible effects of stem cell therapy on the oxidative stress and inflammatory status after acute haloperidol administration in mice with a view to possibly reducing the side effects of the classic antipsychotic.<\/p>\n

Materials and Methods<\/strong><\/p>\n

Animals<\/strong><\/p>\n

Studies were conducted using Swiss male albino mice 22\u201325 g of body weight and aged 5\u2013 6 weeks. Mice were obtained from the animal house colony of the National Research Centre (Cairo, Egypt), housed on a 12-h light\/dark cycle and allowed free access to standard laboratory food and tap water. Animal procedures were done according to the Ethics Committee of the National Research Centre and the recommendations of the National Institutes of Health Guide for Care and Use of Laboratory Animals (Publication No. 85-23, revised 1985).<\/p>\n

Drugs and Chemicals<\/strong><\/p>\n

Haloperidol was obtained from Kahira Pharm and Chem. IND (Cairo, Egypt) and dissolved in sterile physiological saline. Stem cell isolates were obtained from Cairo University. All other chemicals were of analytical grade and purchased from Sigma.<\/p>\n

Study Design<\/strong><\/p>\n

Mice were randomly divided into 4 equal groups (6 mice each). The first 3 groups were treated with haloperidol (5 mg\/kg, i.p., 0.1 ml) for 3 consecutive days and then received either physiological saline (0.2 ml) or stem cell suspension (2 \u00d7 105<\/sup> BM-SCs, 0.2 ml). The 4th<\/sup> group was treated with only saline (i.e. no haloperidol). Mice were euthanized 4h after stem cell injection by decapitation under ether anesthesia. The brain and liver of each mouse were then quickly removed on ice plates, washed with ice-cold phosphate buffered saline (PBS, pH 7.4), weighed, and then stored at\u221280\u00b0C. The brain and liver tissues were homogenized with 0.1 M phosphate buffer saline at pH 7.4 to give a final concentration of 10 %w\/v for the biochemical assays.<\/p>\n

Stem Cell Isolation<\/strong><\/p>\n

Bone marrow was harvested by flushing the rat tibiae and femurs with Dulbecco\u2019s modified Eagle\u2019s medium supplemented with 10% fetal bovine serum. Nucleated cells were then isolated with a density gradient and re-suspended in a complete culture medium supplemented with 1% penicillin\u2013streptomycin. Cells were incubated at 37\u00b0C in 5% humidified CO2 for a period of 12\u201314 days as a primary culture. Following the development of large colonies, cultures were then washed twice with phosphate buffer saline and the cells trypsinized with 0.25% trypsin in 1mM EDTA for 5 min at 37\u00b0C. After centrifugation cells were re-suspended in serum supplemented medium and incubated in 50 cm2<\/sup> culture flask. The resulting cultures were referred to as first-passage cultures. The identification of cells as being MSCs was based on morphology, adherence and the ability to differentiate into osteocytes and chondrocytes.31<\/sup><\/p>\n

Determination of Lipid Peroxidation<\/strong><\/p>\n

Lipid peroxidation was assayed by measuring the level of malondialdehyde in supernatants using the method of Ruiz-Larrea et al.32<\/sup>\u00a0In this assay thiobarbituric acid reactive substances (TBARS) react with thiobarbituric acid to produce a red-colored complex having peak absorbance at 532 nm.<\/p>\n

Determination of Reduced Glutathione<\/strong><\/p>\n

Reduced glutathione was measured in tissue supernatants using the method of Ellman.33<\/sup>\u00a0In this assay, Ellman\u00b4s reagent is reduced by \u2013SH groups of GSH to form 2-nitro-s-mercaptobenzoic acid. The nitromercaptobenzoic acid anion with an intense yellow color can then be determined spectrophotometrically.<\/p>\n

Nitrite Determination<\/strong><\/p>\n

The measurement of nitric oxide in supernatants was done according to the method of Moshage et al.34<\/sup>. In this assay nitrite is used as indicator for the production of nitric oxide.<\/p>\n

Determination of Paraoxonase Activity<\/strong><\/p>\n

Arylesterase activity of paraoxonase was measured spectrophotometrically in supernatants using phenylacetate as a substrate. Arylesterase\/paraoxonase catalyzes the cleavage of phenyl acetate resulting in phenol formation. The rate of formation of phenol is measured by monitoring the increase in absorbance at 270 nm at 25\u00b0C.35,36<\/sup><\/p>\n

Quantification of IL-1\u03b2<\/strong><\/p>\n

Interleukin-1beta (IL-1\u03b2) was measured in tissue supernatants using commercially available IL-1 beta ELISA kit (KOMA BIOTECH Inc.) according to manufacture instructions. The kit uses a double antibody sandwich enzyme linked immunosorbent assay to measure the level of IL-1\u03b2.<\/p>\n

Quantification of IL-6<\/strong><\/p>\n

Interleukin-6 (IL-6) was assayed in tissue supernatants using a double-antibody sandwich enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN).<\/p>\n

Quantification of TNF-\u03b1<\/strong><\/p>\n

The level of TNF-\u03b1 in tissue supernatants was determined with a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit supplied by Thermofischer Scientific Co, USA according to the manufacture instructions.<\/p>\n

Histopathology<\/strong><\/p>\n

Brain and liver sections were fixed in freshly prepared 10 % neutral buffered formalin, processed routinely, and embedded in paraffin. Paraffin sections (5 mm thick) were prepared and stained with haematoxylin and eosin (H&E) for the histopathological examination. Sections were examined using a light microscope (Nikon, Japan).<\/p>\n

Immunohistochemistry<\/strong><\/p>\n

For immunohistochemistry 4 \u03bcm thick deparaffinized tissue sections were used. Briefly, deparaffinized slices were incubated overnight with antibodies against inducible TNF-\u03b1 (diluted 1:100). Endogenous peroxidase activity was blocked by incubation in 0.075% hydrogen peroxide in PBS. For antibody detection DAKO EnVision+ System, Peroxidase\/DAB kit was used. The sections were counterstained with haematoxylin, dehydrated with graded alcohols and xylene, and mounted on slides. The immunostaining intensity and cellular localization TNF-\u03b1 was studied using light microscopy.<\/p>\n

Statistical Analysis<\/strong><\/p>\n

Data are presented as mean \u00b1 SE. The data are analyzed by one way analysis of variance, followed by Duncan\u2019s multiple range test for post hoc comparison of group means. Effects with a probability of p<0.05 are considered significant.<\/p>\n

Results<\/strong><\/p>\n

Biochemical Results<\/strong><\/p>\n

Oxidative Stress Markers<\/strong><\/p>\n

Mice treated with haloperidol alone had significantly higher brain malondialdehyde and nitric acid contents by 35.1% and 48.2% compared with the saline control group. There was a 28.7% and 36.1% decrease in reduced glutathione PON-1 activity, respectively. In addition, haloperidol treatment caused significant increase in the liver malondialdehyde and nitric oxide levels by 60.5% and 58.4%, respectively. Significantly decreased reduced glutathione by 36% and PON-1 activity by 32.4% were also observed in the liver of haloperidol injected mice compared with the saline group. The administration of BM-SCs had no significant effect on the concentration of malondiadehyde, nitric oxide, reduced glutathione in the brain and liver tissue of haloperidol treated mice.\u00a0 BM-SCs administered after haloperidol treatment, however, decreased brain PON-1 activity by 18.2% compared with haloperidol only treatment (Table 1).<\/p>\n

Table 1: Effect of single injection of bone marrow derived stem cells (BM-SCs) on oxidative stress in brain of mice treated with haloperidol (5 mg\/kg) for 3 consecutive days.<\/strong><\/p>\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/td>\nSaline<\/strong><\/td>\nHaloperidol<\/strong><\/td>\nHaloperidol + BM-SCs<\/strong><\/td>\n<\/tr>\n
MDA (nmol\/g. tissue)<\/td>\n25.1 \u00b1 0.96<\/td>\n33.9 \u00b1 1.25*<\/sup><\/td>\n32.7 \u00b1 2.00*<\/sup><\/td>\n<\/tr>\n
Nitric oxide (mmol\/g. tissue)<\/td>\n22.0 \u00b1 1.43<\/td>\n32.6 \u00b1 2.4*<\/sup><\/td>\n32.9 \u00b1 2.1*<\/sup><\/td>\n<\/tr>\n
GSH (mmol\/g. tissue)<\/td>\n4.08 \u00b1 0.34<\/td>\n2.91 \u00b1 0.11*<\/sup><\/td>\n3.11 \u00b1 0.19*<\/sup><\/td>\n<\/tr>\n
PON1 (kU\/l)<\/td>\n14.35 \u00b1 0.93<\/td>\n9.17 \u00b1 0.51*<\/sup><\/td>\n7.50 \u00a0\u00b1 0.82*+<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Results are presented as mean \u00b1 SEM. *p<0.05 vs. saline control group<\/p>\n

Table 2: <\/strong>Effect of single injection of bone marrow derived stem cells (BM-SCs) on oxidative stress in liver of mice treated with haloperidol (5 mg\/kg) for 3 consecutive days. <\/strong><\/p>\n\n\n\n\n\n\n\n
\u00a0<\/strong><\/td>\nSaline<\/strong><\/td>\nHaloperidol<\/strong><\/td>\nHaloperidol + BM-SCs<\/strong><\/td>\n<\/tr>\n
MDA (nmol\/g. tissue)<\/td>\n39.5 \u00b1 1.5<\/td>\n68.38 \u00b1 3.9*<\/sup><\/td>\n64.8 \u00b1 2.3*<\/sup><\/td>\n<\/tr>\n
Nitric oxide (mmol\/g. tissue)<\/td>\n43.8 \u00b1 3.0<\/td>\n69.4 \u00b1 5.6*<\/sup><\/td>\n71.6 \u00b1 3.2*<\/sup><\/td>\n<\/tr>\n
GSH (mmol\/g. tissue)<\/td>\n9.46 \u00b1 0.68<\/td>\n6.06 \u00b1 0.41*<\/sup><\/td>\n5.78 \u00b1 0.65*<\/sup><\/td>\n<\/tr>\n
PON1 (kU\/l)<\/td>\n39.35 \u00b1 2.23<\/td>\n26.60 \u00b1 1.52*<\/sup><\/td>\n30.1 \u00b1 1.90*<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Results are presented as mean \u00b1 SEM. *p<0.05 vs. saline control group<\/p>\n

Inflammatory markers<\/strong><\/p>\n

The concentrations of IL-1\u03b2, IL-6, and TNF-\u03b1 were markedly and significantly elevated in the brain of haloperidol treated mice by 5.11.6%, 344.3% and 249.7%, respectively, compared with the saline group. The administration of BM-SCs significantly reduced IL-1\u03b2, IL-6, and TNF-\u03b1 concentrations by 21.9%, 16.9%, 30.2%, respectively, compared with the haloperidol treated group (Table 2).<\/p>\n

Table 3:\u00a0Effect of single injection of bone marrow derived stem cells (BM-SCs) on brain IL-1\u03b2, IL-6 and TNF-\u03b1 in mice treated with haloperidol (5 mg\/kg) for 3 consecutive days.<\/strong><\/p>\n\n\n\n\n\n\n
\u00a0<\/strong><\/td>\nSaline<\/strong><\/td>\nHaloperidol<\/strong><\/td>\nHaloperidol + BM-SCs<\/strong><\/td>\n<\/tr>\n
IL-1\u03b2 (Pg\/ml)<\/td>\n61.00 \u00b1 2.70<\/td>\n373.11 \u00b1 8.92*<\/sup><\/td>\n291.52 \u00b1 7.43*+<\/sup><\/td>\n<\/tr>\n
IL-6 (Pg\/ml)<\/td>\n4.40 \u00b1 0.23<\/td>\n19.55 \u00b1 0.81*<\/sup><\/td>\n16.25 \u00b1 0.53*+<\/sup><\/td>\n<\/tr>\n
TNF-\u03b1 (Pg\/ml)<\/td>\n18.30 \u00b1 0.76<\/td>\n64.00 \u00b1 3.80*<\/sup><\/td>\n44.70 \u00b1 0.25*+<\/sup><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Results are presented as mean \u00b1 SEM. *p<0.05 vs. saline control group. +*p<0.05 vs. haloperidol control group.<\/p>\n

Histopathological Results<\/strong><\/p>\n

Brain Tissue<\/strong><\/p>\n

In the cortex and striatum of the saline-treated control group the normal structure and neuronal cells with prominent nuclei were observed (Fig.1 & 2 A). <\/strong>Haloperidol caused disorganization of cortical layers, degeneration, shrinkage of neurons, neuronal necrosis, vacuolation of neuropil, eosinophilic areas and pyknosis with apoptotic cells in the cortex and striatum. Perivascular space was found to be increased or dilated along with focal cerebral hemorrhage inside the brain parenchyma (Fig. 1, 2 B & C). However, light microscopic examination of these brain regions after treatment with stem cell suspension, showed return of brain tissues towards normal morphology as evidenced by remarkable regression of the total degenerative changes induced by haloperidol, although pyknosis of some neurons was still seen (Fig. 1, 2 D).<\/p>\n\n\n\n
\"Figure<\/td>\nFigure 1:<\/strong> Representative light microphotographs from the cerebral cortex of mice treated with; (A) Saline showing normal neurons (N) and prominent nuclei (H & E X 400).<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

(B) Haloperidol: cytoplasmic vacuoletion (V), degeneration of neural cells (arrow), and pyknotic darkly stained nuclei (P) with apoptotic cells (arrowhead)(H & E X 400). (C) Haloperidol group: cytoplasmic vacuoletion (V), degeneration of neural cells (arrow), and pyknotic darkly stained nuclei (P) with apoptotic cells (arrowhead).(H & E X 1000). (D) Haloperidol and stem cells: few degenerated neurons (arrow), hemorrhage (H) with few\u00a0 pyknotic nuclei (P) (H & E X 400)<\/p>\n\n\n\n
\"Figure<\/td>\nFigure 2<\/strong>: Representative light microphotographs from the striatum of mice treated with; (A) Saline: normal neurons (N) and prominent nuclei (H & E X 400).<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

(B) Haloperidol: vacuolation of neuropil (V), red neurons (R), pyknotic darkly (arrow) stained and apoptotic nuclei (arrowhead). (C) Haloperidol: shrunken with vacuolation of neuropil (V) and red neurons (R). Pyknotic darkly (arrow) stained and apoptosis nuclei (arrowhead) (H & E X 1000). (D) Haloperidol and stem cells: few pyknotic (arrow) and apoptosis nuclei (arrowhead) (H & E X 400).<\/p>\n

Liver Tissue<\/strong><\/p>\n

Sections from saline treated mice showed normal hepatic architecture with distinct hepatic cells, central vein, sinusoidal spaces and prominent nuclei (Fig.3A). Haloperidol resulted in distortion in hepatocyte arrangement, obvious centrilobular necrosis with degeneration, vacuolation with the nuclei appearing pyknotic in these cells. Inflammatory cell infiltration especially in the periportal area, dark eosinophilic cytoplasm, dilation and sinusoidal hemorrhage with activation of Kupffer cell were also observed (Fig. 3B & C). In the group treated with haloperidol and stem cell suspension, there was almost normalization of the liver tissue and normalized cellular arrangement around the central vein, reduced necrosis with mild dilatation of sinusoids and no congestion. The nuclei of the liver were nearly normal indicating the recovery of the liver tissues, except few pyknotic cells (Fig. 3 D).<\/p>\n\n\n\n
\"Figure<\/td>\nFigure 3: <\/strong>Representative light microphotographs from the liver of mice treated with; (A) Saline: normal structure of the hepatic lobule:\u00a0 central vein (CV) and surrounding hepatocytes (H), sinusoids (S) and nucleus (N).<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

(B) Haloperidol: distortion of hepatic architecture, necrosis with degeneration of hepatocytes with pyknotic nuclei. Dilation and hemorrhage of sinusoids. (C) Haloperidol group: distortion of hepatic architecture, inflammatory cell infiltration especially in the periportal area, dark eosinophilic cytoplasm, degeneration of hepatocytes with pyknotic nuclei. (D) Haloperidol and stem cells: normal hepatic cells with dilatation of hepatic sinusoids (S) and pyknotic nuclei (P) (H & E X 400)<\/p>\n

Tumour Necrosis Factor Immunoreactivity<\/strong><\/p>\n

The cerebral cortex and striatum of the saline control group showed negligible staining for TNF-\u03b1 expression (Figs.\u00a0 4 D & G). Strong TNF-\u03b1 immunoreactivity in the cytoplasm and nuclei of neurons was, however, seen in mice treated with haloperidol (Figs.\u00a0 4 E & H). TNF-\u03b1 expression decreased to nearly normal levels in the haloperidol and stem cells-treated group as compared to that of the haloperidol only group. Only a few scattered TNF-\u03b1-positive cells were observed (Fig. 4\u00a0 F & I).<\/p>\n

In the liver tissue TNF-\u03b1 immunoreactivity was not detected in saline control group (Fig. 4 A). In the haloperidol group, strong TNF-\u03b1 expression was seen mainly in the necrotic areas and predominantly in pericentral and periportal areas (Fig. 4B). Mice treated with haloperidol and stem cells showed markedly reduced TNF-\u03b1 immunoreactivity (Fig. 4C).<\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 4:<\/strong> Representative Immunohistochemical microphotographs of TNF-\u03b1 immunoreactivity from the liver (A\u2013C), cerebral cortex (D\u2013F) and striatum (G\u2013I) of mice treated with saline, haloperidol or haloperidol + stem cell suspension.<\/strong><\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

A\u2013C: liver tissue: (A) Saline-treated mice: there was no TNF-\u03b1 expression. (B) Haloperidol treatment: strong TNF-\u03b1 immunoreactivity. (C) Haloperidol + stem cells: markedly reduced TNF-\u03b1\u00a0 immunoreactivity.<\/p>\n

D\u2013F: cerebral cortex: (A) Saline-treated mice: no TNF-\u03b1 immunoreactivity. (B) Haloperidol treatment: TNF-\u03b1 immunopositivity was strongly increased. (C) Haloperidol + stem cells: few TNF-\u03b1-immunoreactive neurons.<\/p>\n

G\u2013I: striatum: (A) Saline-treated mice: no TNF-\u03b1 immunoreactivity. (B) Haloperidol treatment: strong TNF-\u03b1 immunoreactivity. (C) Haloperidol + stem cells: few TNF-\u03b1-immunoreactive neurons. (TNF-\u03b1, hematoxylin counterstain \u00d7400 and brown color indicates TNF-positivity).<\/p>\n

Discussion<\/strong><\/p>\n

In this study, haloperidol given at 5 mg\/kg for three consecutive days produced neuronal necrosis and apoptosis in the cerebral cortex and striatum as well as hepatic centrilobular necrosis with inflammatory cell infiltration and sinusoidal haemorrhage. We also found that single systemic administration of BM-MSCs suspension was able to ameliorate brain and liver tissue damage caused by acute haloperidol injection in mice. The study thus provides further evidence for a neuro- and hepatic protective effect for MSCs. The administration of haloperidol in the dose of 5 mg\/kg was found to cause lipid peroxidation indicated by the rise in tissue malondialdehyde level. There was also depletion of the intracellular antioxidant and the free radical scavenger reduced glutathione which is likely to reflect its consumption by the increased generation of free radicals. The results obtained are in agreement with other studies indicating increased brain and liver oxidative stress after haloperidol administration in rodents.5,9,7,10,37<\/sup>\u00a0In these studies, chronic haloperidol administration caused increased malondialdehyde and depletion of reduced glutathione in several brain regions and in the liver.7,9<\/sup>\u00a0The drug also increased superoxide in striatum37<\/sup> and decreased brain superoxide dismutase activity,12,38<\/sup>\u00a0glutathione peroxidase activity12<\/sup>, catalase activity and total antioxidant capacity.10<\/sup>\u00a0In vitro, haloperidol increased the levels of TBARS when incubated with plasma from healthy volunteers.39<\/sup>\u00a0In this study, we also found increased brain and liver tissue nitric oxide by haloperidol. These observations confirm other studies that showed increased nitric oxide increased in whole brain and cortex after chronic treatment with haloperidol in mice and rats.7,10<\/sup>\u00a0The increased tissue nitric oxide by haloperidol could be involved in the observed tissue damage via <\/em>the formation of more reactive oxygen and nitrogen species by reacting with molecular oxygen. When present in high amounts, nitric oxide can react with the superoxide anion (O2<\/sub>\u00b7-<\/sup>) resulting in the highly reactive peroxynitrite (ONOO\u2212) capable of oxidation of thiols or protein thiols or nitration of protein tyrosine residues. Other reactive nitrogen oxides such as nitrogen dioxide (\u2022<\/sup>NO2<\/sub>) and dinitrogen trioxide (N2<\/sub>O3<\/sub>) can nitrosate thiols or protein cysteine residues.40<\/sup><\/p>\n

The results from this study also support previous work indicating significantly decreased paraoxonase-1 (PON-1) activity in brain and liver after haloperidol treatment in rats.10<\/sup>\u00a0The enzyme PON-1 which is synthesized by the liver is involved in the detoxification of some organophosphate insecticides and nerve agents.41<\/sup>\u00a0Moreover, PON-1 has been found to have an antioxidant and anti-inflammatory roles36,42<\/sup> and decreased activity was reported in such conditions as such as\u00a0 dementia,43<\/sup>\u00a0Alzheimer\u2019s disease44\u00a0<\/sup>and autism.45<\/sup>\u00a0Plasma serum arylesterase activity also fell in first episode schizophrenic patients.46,47<\/sup>\u00a0Paraoxonase-1 is sensitive to oxidative stress48<\/sup> and decreased activity might thus reflect inactivation by reactive oxygen metabolites.<\/p>\n

The present work show that the acute haloperidol treatment results in neuronal necrosis and apoptosis and eosinophilic areas in the cerebral cortex and striatum of mice. Increased number of apoptotic cells in the striatum has been reported following administration of a single dose of 4mg\/kg haloperidol in rats. The higher dose of 12 mg\/kg resulted in increased number of apoptotic cells in both the striatum and substantia nigra pars reticulata.11<\/sup>\u00a0In vitro treating HT-22 cells with haloperidol resulted in increased number of apoptotic cells (TUNEL staining) and increased the expression of pro-apoptotic (Bax) protein in the hippocampus and caudate putamen of treated rats.49<\/sup>\u00a0Neuronal atrophy and necrosis in the cerebral cortex and sunstantia nigra has also been reported following chronic haloperidol treatment at a dose of 2 mg\/kg for 21 days.12<\/sup>\u00a0Haloperidol thus causes neuronal damage which could be due to increased oxidative\/nitrosative burden. The latter process can result in consequent impairment of mitochondrial respiration and brain energy depletion.50,51<\/sup>\u00a0Our findings are important in view of the data implicating increased oxidative stress in schizophrenia.13,14<\/sup>\u00a0In these patients there is increased malondialdehyde46<\/sup> and F2-isoprostane levels52<\/sup> in plasma as well as increased urinary excretion of RNA and DNA oxidation products.53<\/sup>\u00a0Studies have also shown significantly higher malondialdehyde levels in the plasma of schizophrenia patients treated with first generation antipsychotics like haloperidol compared to second generation agents.54<\/sup><\/p>\n

There is ample evidence that suggests a role for pro-inflammatory cytokines in the pathogenesis of schizophrenia. Thus, first episode schizophrenia showed increased serum or plasma IL-1\u03b2,55<\/sup>\u00a0IL-6, TNF-\u03b1,18,19,47<\/sup>\u00a0IL-447<\/sup> and IL-1856<\/sup> in first episode schizophrenia patients compared with healthy controls. Schizophrenia patients with acute exacerbation of exhibited increased plasma IL-6 levels compared with patients in remissions.57<\/sup>\u00a0In addition, increased IL-6 mRNA in mononuclear cells from peripheral blood has also been demonstrated in schizophrenics with worse positive symptoms.20<\/sup>\u00a0In this study, the concentrations of IL-1\u03b2, IL-6 and TNF-\u03b1 were measured in brain tissue of mice after treatment with haloperidol. We found significantly increased concentrations of the studied pro-inflammatory cytokines compared to the control group. We also demonstrated strong TNF-\u03b1 immunoreactivity in the cerebral cortex and striatum of haloperidol treated mice compared to controls. Haloperidol might thus be able to exacerbate the inflammatory process in the schizophrenic brain and consequently the progression to neurodegeneration.<\/p>\n

In this study, the systemic administration of BM-MSCs caused remarkable regression of the neurotoxic and hepatotoxic effects of haloperidol as evidenced by the histologic evaluation of brain and liver tissue after BM-MSCs treatment. In both the brain and liver tissue of haloperidol-treated mice, lipid peroxidation, nitric oxide or reduced glutathione were not altered by BM-MSCs treatment. Brain PON-1 activity, however, showed further decrease after stem cell therapy. Thus, the protective effect of BM-MSCs was not due to an antioxidant action.\u00a0 Stem cell therapy, however, exerted anti-inflammatory action causing significant decrease in the levels of the proinflammatory cytokines IL-1\u03b2, IL-6 and TNF-\u03b1 in brain of haloperidol treated mice. Our results are therefore in favor for an anti-inflammatory action for stem cell suspension in decreasing haloperidol-induced neuronal and liver damage. In previous studies, i.p. MSCs of bone marrow or adipose tissue origin were shown to ameliorate neuronal damage in different brain regions and also hepatocyte degeneration and liver inflammation caused by lipopolysaccharide (LPS) endotoxin in mice. The oxidative stress response was not reduced but there were decreased expression of caspase-3, cyclooxygenase-2, TNF-\u03b1 and inducible nitric oxide synthase in brain and liver tissue by MSCs treatment.58,59\u00a0<\/sup>MSCs injected i.v. were able to ameliorate the behavioral changes and the loss in tyrosine hydroxylase positive neurons in the substantia nigra pars compacta in an experimental model of Parkinson\u2019s disease via an anti-apoptotic action.26<\/sup>\u00a0In experimental traumatic brain injury, i.v. MSCs reduced the release of cytokines into the brain tissue.25\u00a0<\/sup>In a model of fulminant hepatic failure, i.v. MSCs reduced mortality, hepatocyte dealth and tissue leucocytic infiltration.27<\/sup>\u00a0MSCs injected i.v. were also shown to modulate the inflammatory response in renal injury reducing the expression levels of IL-6, TNF-\u03b1 mRNA and decreasing serum IL-1\u03b1, IL-1\u03b2, INF-g and INF-\u03b1.\u00a060\u00a0<\/sup>It is thus becoming clear that MSCs exert their therapeutic effects via an anti-apoptotic action and by modulating the inflammatory response.<\/p>\n

In summary, the present study demonstrate that BM-MSCs given i,p. can protect against brain injury and hepatocyte death caused by short term haloperidol treatment in high dose in mice. This therapeutic benefit of BM-MSC is likely to be mediated by interfering with the inflammatory response.<\/p>\n

Acknowledgements<\/strong><\/p>\n

This works is was not supported by research grants<\/p>\n

Conflicts of Interest<\/strong><\/p>\n

There is no conflict of interest.<\/p>\n

References<\/strong><\/p>\n

    \n
  1. Miller B. J., Buckley P., Seabolt W., Mellor A., Kirkpatrick B. Meta-analysis of cytokine alterations in schizophrenia: clinical status and antipsychotic effects. Biol Psychiatry.\u00a0<\/em> 2011;70:663\u2013671.
    \n    CrossRef<\/a><\/li>\n
  2. Labarca R., Silva H., Jerez S., Ruiz A., Forray M. I., Gysling K., Andres M. E., Bustos G., Castillo Y., Hono J.\u00a0 Differential effects of haloperidol on negative symptoms in drug-naive schizophrenic patients: effects on plasma homovanillic acid. Schizophr Res.<\/em> 1993;9(1):29-34.
    \n    CrossRef<\/a><\/li>\n
  3. Miyamoto S., Duncan G. E., Marx C. E., Lieberman J. A. Treatments for schizophrenia a critical review of pharmacology and mechanisms of action of antipsychotic drugs. Molecular Psychiatry.\u00a0<\/em> 2005;10:79\u2013104.
    \n    CrossRef<\/a><\/li>\n
  4. Meltzer H. Y., Park S., Kessler R. Cognition, schizophrenia, and the atypical antipsychotic drugs. Proc Natl Acad Sci U S A.<\/em> 1999;96(24):13591\u201313593.
    \n    CrossRef<\/a><\/li>\n
  5. Post A., Holsboer F., Behl C. Induction of NF-kB activity during haloperidol-induced oxidative toxicity in clonal hippocampal cells: suppression of NF-kB and neuroprotection by antioxidants. J Neurosci.\u00a0<\/em> 1998;18(20):8236\u20138246.
    \n    CrossRef<\/a><\/li>\n
  6. Martins M. R., Petronilho F. C., Gomes K. M., Dal-Pizzol F., Streck E. L., Quevedo J.\u00a0 Antipsychotic-induced oxidative stress in rat brain. Neurotox Res.<\/em> 2008;13:63-9.
    \n    CrossRef<\/a><\/li>\n
  7. Abdel-Salam O. M., El-Sayed El-Shamarka M., Salem N. A., El-Mosallamy A. E., Sleem A. A. Amelioration of the haloperidol-induced memory impairment and brain oxidative stress by cinnarizine. EXCLI J.\u00a0<\/em> 2012;11:517-30.<\/li>\n
  8. Halliwell B., Gutteridge J. M. C. Free radicals in biology and medicine, 3rd edn. Clarendon Press, Oxford. 199.<\/li>\n
  9. Vairetti M., Feletti F., Battaglia A., Pamparana F., Canonico P. L., Richelmi P., Bert\u00e8 F. Haloperidol-induced changes in glutathione and energy metabolism: effect of nicergoline. Eur J Pharmacol<\/em>. 1999;367:67-72.
    \n    CrossRef<\/a><\/li>\n
  10. Abdel-Salam O. M., Youness E. R., Khadrawy Y. A., Sleem A. A. Brain and liver oxidative stress after sertraline and haloperidol treatment in mice. J Basic Clin Physiol Pharmacol.<\/em>\u00a0 2013;24(2):115-23.
    \n    CrossRef<\/a><\/li>\n
  11. Mitchell I. J., Cooper A. C., Griffiths M. R., Cooper A. J.\u00a0 Acute administration of haloperidol induces apoptosis of neurones in the striatum and substantia nigra in the rat. Neuroscience.<\/em> 2002; 109(1):89-99.
    \n    CrossRef<\/a><\/li>\n
  12. Perera J., Tan J. H., Jeevathayaparan S., Chakravarthi S., Haleagrahara N.\u00a0 Neuroprotective effects of alpha lipoic acid on haloperidol-induced oxidative stress in the rat brain. Cell Biosci<\/em>. 2011;1(1):12.
    \n    CrossRef<\/a><\/li>\n
  13. Wu J. Q., Kosten T. R., Zhang X. Y.\u00a0 Free radicals, antioxidant defense systems, and schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry<\/em> .2013;46:200-206.
    \n    CrossRef<\/a><\/li>\n
  14. Flatow J., Buckley P., Miller B. J.\u00a0 Meta-analysis of oxidative stress in schizophrenia. Biol Psychiatry<\/em>.\u00a0 2013;74:400\u2013409.
    \n    CrossRef<\/a><\/li>\n
  15. Berk M., Copolov D., Dean O., Lu K., Jeavons S., Schapkaitz I., Anderson-Hunt M., Judd F., Katz F., Katz P., Ording-Jespersen S., Little J., Conus P., Cuenod M.,\u00a0 KQ D., Bush A. I.\u00a0 N-acetyl cysteine as a glutathione precursor for schizophrenia-a double-blind, randomized placebo-controlled trial.Biol Psychiatry<\/em>.\u00a0 2008;64(5):361-8.
    \n    CrossRef<\/a><\/li>\n
  16. Dean O., Giorlando F., Berk M.\u00a0 N-acetylcysteine in psychiatry: current therapeutic evidence and potential mechanisms of action. J Psychiatry Neurosci<\/em>. 2011;36(2):78-86.
    \n    CrossRef<\/a><\/li>\n
  17. Bo\u0161kovi\u0107 M., Vovk T., Kopriv\u0161ek J., Plesni\u010dar B. K., Grabnar I.\u00a0 Vitamin E and essential polyunsaturated fatty acids supplementation in schizophrenia patients treated with haloperidol. Nutr Neurosci.<\/em> 2016;19(4):156-61.
    \n    CrossRef<\/a><\/li>\n
  18. O’Brien S. M., Scully P., Dinan T. G. Increased tumor necrosis factor-alpha concentrations with interleukin-4 concentrations in exacerbations of schizophrenia. Psychiatry Res<\/em>. 2008;160(3):256-62.
    \n    CrossRef<\/a><\/li>\n
  19. Kubistova A., Horacek J., Novak T.\u00a0 Increased interleukin-6 and tumor necrosis factor alpha in first episode schizophrenia patients versus healthy controls. Psychiatria Danubina.<\/em> 2012;24(1):153\u2013156.<\/li>\n
  20. Chase K. A., Cone J. J., Rosen C., Sharma R. P. The value of interleukin 6 as a peripheral diagnostic marker in schizophrenia. BMC Psychiatry<\/em>. 2016;16:152.
    \n    CrossRef<\/a><\/li>\n
  21. Weiner L. P. Definitions and criteria for stem cells. Methods Mol Biol.<\/em>\u00a0 2008;438:3-8.
    \n    CrossRef<\/a><\/li>\n
  22. Verfaillie C. M.\u00a0 Multipotent adult progenitor cells: an update. Novartis Found Symp<\/em>.\u00a0 2005;265:55-61.
    \n    CrossRef<\/a><\/li>\n
  23. Ramsay M. A. E.\u00a0 Will stem cells transform medicine? Proc (Bayl Univ Med Cent)<\/em>.\u00a0 2002;15:135\u2013137
    \n    CrossRef<\/a><\/li>\n
  24. Zhang R., Liu Y., Yan K., Chen L., Chen X. R., Li P., Chen F. F., Jiang X. D.\u00a0 Anti-inflammatory and immunomodulatory mechanisms of mesenchymal stem cell transplantation in experimental traumatic brain injury. J Neuroinflammation<\/em>.\u00a0 2013;10:106.
    \n    CrossRef<\/a><\/li>\n
  25. Galindo L. T., Filippo T. R. M.,Semedo P., Ariza C. B., Moreira C. M., Camara N. O. S., Porcionatto M. A.\u00a0 Mesenchymal stem cell therapy modulates the inflammatory response in experimental traumatic brain injury. Neurol Res Int.\u00a0<\/em> 2011:564089.
    \n    CrossRef<\/a><\/li>\n
  26. Wang F., Yasuhara T., Shingo T., Kameda M., Tajiri N., Yuan W. J., Kondo A., Kadota T., Baba T., Tayra J. T., Kikuchi Y., Miyoshi Y., Date I.\u00a0 Intravenous administration of mesenchymal stem cells exerts therapeutic effects on parkinsonian model of rats focusing on neuroprotective effects of stromal cell-derived factor-1alpha. BMC Neurosci<\/em> 2010;11:52.
    \n    CrossRef<\/a><\/li>\n
  27. Parekkadan B., van Poll D., Suganuma K., Carter E. A., Berthiaume F., Tilles A. W., Yarmush M. L. Mesenchymal stem cell-derived molecules reverse fulminant hepatic failure. PLoS One.<\/em> 2007;2(9):e941.
    \n    CrossRef<\/a><\/li>\n
  28. Zhao W., Li J. J., Cao D. Y., Li X., Zhang L. Y., He Y., Yue S. Q., Wang D. S., Dou K. F. Intravenous injection of mesenchymal stem cells is effective in treating liver fibrosis. World J Gastroenterol.<\/em> 2012;18(10):1048-1058.
    \n    CrossRef<\/a><\/li>\n
  29. Ya\u00f1ez R., Lamana M. L., Garc\u00eda-Castro J., Colmenero I., Ram\u00edrez M., Bueren J .A.\u00a0 Adipose tissue-derived mesenchymal stem cells have in vivo immunosuppressive properties applicable for the control of the graft-versus-host disease. Stem Cells.<\/em> 2006;24(11):2582-91.
    \n    CrossRef<\/a><\/li>\n
  30. Cao H., Qian H., Xu W., Zhu W., Zhang X., Chen Y., Wang M., Yan Y., Xie Y.\u00a0 Mesenchymal stem cells derived from human umbilical cord ameliorate ischemia\/reperfusion-induced acute renal failure in rats. Biotechnol Lett.\u00a0<\/em> 2010;32(5):725-32.
    \n    CrossRef<\/a><\/li>\n
  31. Aziz M. T. A., Wassef M. A., Rashed L. A., Mhfouz S., Omar N., Elsebaie M. M.\u00a0 Mesenchymal stem cells therapy in acute renal failure: possible role of hepatocyte growth factor. J Stem Cell Res Ther<\/em>. 2011; 1:3.<\/li>\n
  32. Ruiz-Larrea M. B., Leal A. M., Liza M., Lacort M., de Groot H.\u00a0 Antioxidant effects of estradiol and 2-hydroxyestradiol on iron induced lipid peroxidation of rat liver microsomes. Steroids.<\/em> 1994; 59:383\u2013388.
    \n    CrossRef<\/a><\/li>\n
  33. Ellman G. L.\u00a0 Tissue sulfhydryl groups. Arch Biochem<\/em>.\u00a0 1959;82:70\u201377.
    \n    CrossRef<\/a><\/li>\n
  34. Moshage H., Kok B., Huizenga J. R.\u00a0 Nitrite and nitrate determination in plasma a critical evaluation. Clin Chem.<\/em>\u00a0 1995;41:892\u2013896<\/li>\n
  35. Higashino K., Takahashi Y., Yamamura Y.\u00a0 Release of phenylacetate esterase from liver microsomes by carbon tetrachloride. Clin Chim Acta<\/em>.\u00a0 1972;41:313\u2013320.
    \n    CrossRef<\/a><\/li>\n
  36. Watson A. D., Berliner J. A., Hama S. Y., Du B. N. L., Faull K. F., Fogelman A. M., Navab M. Protective effect of high density lipoprotein associated paraoxonase. Inhibition of the biological activity of minimally oxidized low density lipoprotein. J Clin Invest.<\/em>\u00a0 1995;96:2882\u20132891.
    \n    CrossRef<\/a><\/li>\n
  37. Harvey B. H., Joubert C., du Preez J. L., Berk M. Effect of chronic N-acetyl cysteine administration on oxidative status in the presence and absence of induced oxidative stress in rat striatum. Neurochemical Res<\/em>.\u00a0 2008;33(3):508\u2013517
    \n    CrossRef<\/a><\/li>\n
  38. Pillai A., Parikh V., Terry A. V Jr., Mahadik S. P.\u00a0 Long-term antipsychotic treatments and crossover studies in rats: differential effects of typical and atypical agents on the expression of antioxidant enzymes and membrane lipid peroxidation in rat brain. J Psychiatr Res<\/em>. 2007;41:372-86.
    \n    CrossRef<\/a><\/li>\n
  39. Dietrich-Muszalska A., Kontek B., Rabe-Jab\u0142o\u0144ska J.\u00a0 Quetiapine, olanzapine and haloperidol affect human plasma lipid peroxidation in vitro. Neuropsychobiology.<\/em>\u00a0 2011;63(4):197-201.
    \n    CrossRef<\/a><\/li>\n
  40. Thomas D. D., Miranda K. M., Citrin D., Espey M. C., Wink D. A.\u00a0 Nitric Oxide. In: Combat Medicine: Basic and Clinical Research in Military, Trauma, and Emergency Medicine.\u00a0\u00a0 Tsokos G. c., Atkins J. L (Eds) Humana Press Inc., Totowa N. J.\u00a0 2003;23-60.
    \n    CrossRef<\/a><\/li>\n
  41. Du B. N. L.\u00a0 Human serum paraoxonase: arylesterase. In: Kalow W., editor. Pharmacogenetics of Drug Metabolism. New York: Pergamon Press.\u00a0 1992, pp.51\u201391.<\/li>\n
  42. Ng D. S., Chu T., Esposito B., Hui P., Connelly P. W., Gross P. L.\u00a0 Paraoxonase-1 deficiency in mice predisposes to vascular inflammation, oxidative stress, and thrombogenicity in the absence of hyperlipidemia. Cardiovasc Pathol<\/em>. 2008;17(4):226-32.
    \n    CrossRef<\/a><\/li>\n
  43. Wehr H., Bednarska-Makaruk M., Graban A., Lipczynska-Lojkowska W., Rodo M., Bochynska A., Ryglewicz D. Paraoxonase activity and dementia. J Neurol Sci<\/em>. 2009;283(1\u20132):107\u2013108.
    \n    CrossRef<\/a><\/li>\n
  44. Zengi O., Karakas A., Ergun U., Senes M., Inan L., Yucel D.\u00a0 Urinary 8-hydroxy-2\u2019-deoxyguanosine level and plasma paraoxonase 1 activity with Alzheimer\u2019s disease. Clinical Chemistry and Laboratory Medicine.<\/em> 2011;50:529-534.<\/li>\n
  45. Abdel-Salam O. M. E,, Youness E. R., Mohammed N. A., Elhamed W. A. A.\u00a0 Nuclear factor-kappa B and other oxidative stress biomarkers in serum of autistic children. Open J Mol Integr Physiol.<\/em> 2015;5:18-27.
    \n    CrossRef<\/a><\/li>\n
  46. Sarandol A., Sarandol E., Acikgoz H. E., Eker S. S., Akkaya C., Dirican M. First-episode psychosis is associated with oxidative stress: Effects of short-term antipsychotic treatment. Psychiatry Clin Neurosci.<\/em> 2015;69(11):699-707.
    \n    CrossRef<\/a><\/li>\n
  47. Brinholi F. F., Noto C., Maes M., Bonif\u00e1cio K. L., Brietzke E., Ota V. K., Gadelha A., Cordeiro Q., Belangero S. I., Bressan R. A., Vargas H. O., Higachi L., de Farias C. C., Moreira E. G., Barbosa D. S.\u00a0 Lowered paraoxonase 1 (PON1) activity is associated with increased cytokine levels in drug na\u00efve first episode psychosis. Schizophr Res<\/em>. 2015;166(1-3):225-30.
    \n    CrossRef<\/a><\/li>\n
  48. Nguyen S. D., Sok D. E. Preferential inhibition of paraoxonase activity of human paraoxonase 1 by negatively charged lipids. J Lipid Res.<\/em> 2004;45(12):2211\u201320.
    \n    CrossRef<\/a><\/li>\n
  49. Post A., R\u00fccker M., Ohl F., Uhr M., Holsboer F., Almeida O. F., Michaelidis T. M.\u00a0 Mechanisms underlying the protective potential of alpha-tocopherol (vitamin E) against haloperidol-associated neurotoxicity. Neuropsycho. pharmacology<\/em>. 2002;26(3):397-407.
    \n    CrossRef<\/a><\/li>\n
  50. Halliwell B. Oxidative stress and neurodegeneration: where are we now? J Neurochem<\/em>. 2006;97:1634\u20131658.
    \n    CrossRef<\/a><\/li>\n
  51. Brown G. C.\u00a0 Nitric oxide and neuronal death. Nitric Oxide. 2010;23:153\u2013165.
    \n    CrossRef<\/a><\/li>\n
  52. Lee E. E., Eyler L. T., Wolkowitz O. M., Martin A. S., Reuter C., Kraemer H., Jeste D. V. Elevated plasma F2-isoprostane levels in schizophrenia. Schizophr Res<\/em>. 2008;176(2-3):320-6.
    \n    CrossRef<\/a><\/li>\n
  53. Jorgensen A., Broedbaek K., Fink-Jensen A., Knorr U., Soendergaard M. G., Henriksen T., Weimann A., Jepsen P., Lykkesfeldt J., Poulsen H. E., Jorgensen M. B.\u00a0 Increased systemic oxidatively generated DNA and RNA damage in schizophrenia. Psychiatry Res.<\/em> 2013;209(3):417-23.
    \n    CrossRef<\/a><\/li>\n
  54. Kropp S., Kern V., Lange K., Degner D., Hajak G., Kornhuber J., R\u00fcther E., Emrich H. M., Schneider U., Bleich S.\u00a0 Oxidative stress during treatment with first- and second-generation antipsychotics. J Neuropsychiatry Clin Neurosci<\/em>.\u00a0 2005;17(2):227-31.
    \n    CrossRef<\/a><\/li>\n
  55. Miller R. Mechanisms of action of antipsychotic drugs of different classes, refractoriness to therapeutic effects of classical neuroleptics and individual variation in sensitivity to their actions Part II. Curr Neuropharmacol.<\/em> 2009;7(4):315-30.
    \n    CrossRef<\/a><\/li>\n
  56. Xiu M. H., Chen D. C., Wang D., Zhang K., Dong A., Tang W., Zhang F., Liu L. J., Liu J. H., Liu H. B., Yang F. D., Kosten T. R., Zhang X.Y.\u00a0 Elevated interleukin-18 serum levels in chronic schizophrenia: Association with psychopathology. J Psychiatr Res.<\/em> 2012;46(8):1093-1098.
    \n    CrossRef<\/a><\/li>\n
  57. Naudin J., M\u00e8ge J. L., Azorin J. M., Dassa D. Elevated circulating levels of IL-6 in schizophrenia. Schizophr Res.<\/em> 1996;20(3):269-73.
    \n    CrossRef<\/a><\/li>\n
  58. Abdel-Salam O. M. E., Youness E. R., Omara E. A., Sleem A. A.\u00a0 Effect of adipose tissue-derived mesenchymal stem cell treatment on oxidative stress and inflammatory response following Escherichia coli lipopolysaccharide. Comp Clin Patho<\/em>l. 2015;24 (2):343\u2013358.
    \n    CrossRef<\/a><\/li>\n
  59. Abdel-Salam O. M. E., Youness E. R., Omara E. A., El-Sayed., El-Shamarka M., Sleem A. A. Protection by intraperitoneal administration of bone marrow-derived stem cells of lipopolysaccharide-induced brain and liver damage in mice. Comp Clin Pathol<\/em> 2016;25(1):107\u2013118.
    \n    CrossRef<\/a><\/li>\n
  60. Semedo P., Correa-Costa M., Antonio Cenedeze M., Avancini M. C.,Malheiros D.,\u00a0 dos Reis M. A., Shimizu M. H., Seguro A. C., Pacheco-Silva A., Camara N. O. S . Mesenchymal stem cells attenuate renal fibrosis through immune modulation and remodeling properties in a rat remnant kidney model. Stem Cells.<\/em> 2009;27(12):3063-73.
    \n    CrossRef<\/a><\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction Haloperidol is a classic antipsychotic drug in use to […]<\/p>\n","protected":false},"author":9,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[55],"tags":[],"class_list":["post-19546","post","type-post","status-publish","format-standard","hentry","category-vol11no1"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/19546","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/9"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=19546"}],"version-history":[{"count":6,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/19546\/revisions"}],"predecessor-version":[{"id":32083,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/19546\/revisions\/32083"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=19546"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=19546"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=19546"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}