Cyclooxygenase-2 Immunoreactivity Research Articles

Boosting impacts of Acacia nilotica against hepatic toxicity induced by gentamicin: biochemical, anti-inflammatory and immunohistochemical study.

It seems that gentamicin's toxicity to the liver is caused by reactive oxygen species production. The antioxidant and anti-inflammatory properties of Acacia nilotica extract (AN) have been demonstrated in recent studies. This research focused on how AN's extract affected gentamicin-induced liver damage in rats. Twenty-four Wister rats of male type were divided into four groups: first group received saline as a control, second group received AN (5%) for fifteen days, group three received daily intraperitoneal injections of gentamicin (100mg/kg) for fifteen days, and group four, as mentioned in groups 2 and 3, also received gentamicin injections and AN extraction (5%) for fifteen days. In order to conduct biochemical analysis, serum was extracted. Histopathology, immunohistochemistry analyses for hepatic toxicity were all performed on the collected tissue samples. Serum levels of ALT, AST, total bilirubin, and GGT were all elevated after using gentamicin. The inflammatory cytokines)IL-1, TNF-α and IL-6(, all were increased in gentamycin-injected group. There were showing deformity of bile duct, hepatocellular necrosis and infiltration of inflammatory cells congestion of portal vein, and hepatic sinusoids besides fibrosis of portal area (white arrows), hypertrophy in gentamycin-injected group compared to AN plus gentamycin administered rats. There were upregulation in the immunoreactivity of COX-2, IFNkB and TGF-beta1 (TGF-β1) in gentamycin intoxicated rats. When gentamicin and AN were administered together, hepatic biomarkers, inflammatory cytokines, histological, and immunohistochemical markers were all ameliorated by AN administration.

739. SIGNIFICANCE OF COX2 INHIBITOR FOR ESOPHAGEAL CANCER FROM THE CLINICAL AND EXPERIMENTAL ESTIMATE

Abstract (Aims) We report experimental and clinical evidence that COX2 represent a potential molecular target for the treatment and prevention of esophageal cancer. (Methods) (A)Experiment: Thirty 8-week male wistar rats were exposed to duodenal content esophageal reflux. All animal underwent an esophagoduodenal anastomosis (EDA) with total gastrectomy in order to produce chronic esophagitis. In ten rats the sham (Control). They were sacrified at 40th week. Their esophagi were examined for HE, COX2,PGE2, PCNA and total bile acids in the esophageal lumen was measured. (B)Clinical: The expression of COX2 was examined for 68 specimens of esophageal squamous cell carcinoma (SCC) and the correlation of COX2 expression with clinicopatholigic features was examined. (Results) (A) After 40 weeks of reflux, columnar dysplasia, squamous cell carcinoma and adenocarcinoma were found. Total bile acid in the esophageal lumen was significantly increased in the EDA group compared with the sham operated rats. PCNA LI and esophageal tissue PGE2 were higher in dysplastic and cancer tissue than that of control. Overexpression of COX2 was shown in dysplastic and cancer tissue. COX2 may play an important role in esophageal cancer by duodenal content reflux.(B) COX2 immunoreactivity was weak in 27(40%) and strong in 41(60%) of the carcinomas. The proportion of poorly differentiated SCC among tumors with a strong expression of COX2 (34%) was significantly higher than among tumors with a weak expression of COX2 (19%, p=0.02). The depth of tumors (p=0.004), lymph node metastasis (p=0.0009) and the stage of the tumors (p=0.001) were advanced significantly more progressively in ECCS with a strong COX2 expression. Moreover, survival was significantly reduced (p=0.02) among patients with strong COX2 expression when compared with the COX2 weak group. This study showed that strong expression of COX2 was correlated with tumor progression and poor differentiation in ESCC. (Conclusion) In this experimental study, we demonstrate that bile reflux of duodenal contents induce COX2 and increase prostaglandin synthesis in dysplastic and cancer tissue. Our study suggests that COX2 may play a role in esophageal carcinoma development and progression, COX2 inhibitors may be potential targets for the prevention or treatment of esophageal carcinoma.

Kırmızı Reishi Mantarı (Ganoderma lucidum) Extratının Karbon Tetraklorür ile İndüklenen Karaciğer Hasarı ve Siklooksijenaz-2 İmmunoreaktivitesi Üzerine Etkisi

Carbon tetrachloride (CCl4) is a xenobiotic compound with toxicological action. It is absorbed by gastrointestinal system, respiratory system, and skin. Studies have reported that many countries have used Ganoderma lucidum (GL, Reishi Mushroom) as a medicinal mushroom against liver diseases induced by hepatotoxic agents such as CCl4 for more than thousands of years and is used for many diseases, including cancer since it has been thought that it increases resistance against them and treats them. In the present study, immunohistochemical localization and expression of cyclooxygenase-2 (COX-2) by administrating carbon tetrachloride and Ganoderma lucidum in adult rats were examined. In the study, 32 adult Spraque-Dawley male rats that were 8-10 weeks old were used. Rats were divided into 4 groups as control, CCI4, Ganoderma lucidum (GL), and CCI4+GL. As a result of the experimental applications, the liver tissue was found to be normal in the control and GL groups, and multifocal necrosis areas, hepatocellular degeneration, cell infiltration, sinusoidal dilatation, and congestion were observed in the central and portal areas in CCI4 group. In the CCI4+GL group, decreases were observed in lesion severity and density. COX-2 immunoreactivity was detected as more common in hepatocyte cytoplasm in the area from the central vena to the Kiernan space, while it was observed as sporadic in the hepatocyte nucleus. While CCI4 caused a decrease in total antioxidant level (TAS) in blood plasma samples, it caused an increase in total oxidant level (TOS), Aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels. It is seen that Ganoderma lucidum, which has an important place in alternative and folk medicine, reduces oxidative stress with its hepatoprotective effect and inhibits the inflammatory response in the liver.

Ameliorative impacts of Rhodiola rosea against hepatic toxicity induced by monosodium glutamate: role of inflammation-, oxidative-stress-, and apoptosis-associated markers

Current study examined the protective impact of Rhodiola rosea (R. rosea) extract on hepatic toxicity markers caused by MSG. MSG induced significant increases in hepatic stress biomarkers. There were significant declines in GSH, catalase and SOD levels, which were accompanied by substantial elevations in IL-1β, IL-6, and TNF-α. There was hepatic cellular damage, and upregulation of caspase-3 and a decrease in Bcl-2 expression. The hepatotoxic impacts of MSG were normalized by Rhodiola supplementation. R. rosea downregulated caspase-3 and upregulated HO-1 and Nrf2 mRNA expression. R. rosea regulated the immunoreactivity of COX-2, TGF-β1, and NFkB genes associated with hepatic toxicity. R. rosea extract normalized histopathological changes induced by MSG and restored hepatic architecture. R. rosea extract exhibited anti-apoptotic, antioxidant, and anti-inflammatory properties. In conclusion, R. rosea extract is a promising medication against hepatic toxicity induced by MSG, which works by regulating the different signaling pathways of inflammation-, oxidative-stress-, and apoptosis-associated markers.

Cakes fortified with papaya seeds effectively protects against CCl4-induced immunotoxicity

Maintaining a robust immune system and safeguarding the liver from toxins are crucial for overall health. The study aimed to investigate the immunostimulant effects of papaya seed-enriched cakes (CPS) in countering carbon tetrachloride (CCl4)-induced immunocytotoxicity in rats (n = 48). The rats were divided into six groups (8 each): a control group (Group 1), rats fed cakes containing 15% papaya seeds (Group 2 — CPS), rats exposed only to CCl4 (Group 3 — CCl4), rats injected with CCl4 and administered silymarin (Group 4 — CCl4 + S), rats receiving both CCl4 and cakes with papaya seeds (Group 5 — CCl4 + CPS), and rats receiving both CCl4 and silymarin with papaya seed-enriched cakes (Group 6 — CCl4 + CPS + S). HPLC analysis of papaya seeds revealed the presence of ten polyphenol compounds, with quercetin, apigenin, and catechin identified as major flavonoids, along with pyrogallol, ellagic, and gallic acid as predominant phenolic acids. These compounds displayed potent antioxidant activity, attributed to the seeds’ high total phenolic and flavonoid content. The administration of CCl4 significantly affected hematological parameters, liver enzymes, hepatic oxidative stress, levels of TNF-α, IL-6, IgG, as well as IgM. However, rats fed with CPS exhibited mitigation of CCl4-induced toxic effects on hematological parameters and hepatotoxicity. CPS consumption enhanced the antioxidant system, improved inflammatory markers, and immune parameters, restoring them to normal levels. Histopathological analysis confirmed CPS’s ability to reduce CCl4-induced hepatocellular necrosis. Immunohistochemical assessment further revealed reduced immunoreactivity against cleaved caspase-3 expression and increased COX2 immunoreactivity, indicating hepatocellular regeneration in CPS. The combination of CPS and silymarin demonstrated even more notable improvements, suggesting augmented protective impacts against CCl4-induced immunosuppression and hepatotoxicity. In conclusion, CPS exhibited antioxidant properties and effectively protected against CCl4-induced immunotoxicity and hepatotoxicity, with additional benefits observed when combined with silymarin. These findings emphasize the potential health advantages of incorporating papaya seeds into food products, promoting immune system health, and safeguarding against liver damage induced by hazardous agents like CCl4.

Pathological studies on spontaneous urinary bladder lesions in Indian buffaloes

The present investigation describes the spontaneous pathological lesions in urinary bladders (n = 177) of slaughtered buffaloes collected from Northern India. Gross examination revealed congestion, haemorrhages, oedematous changes and thickening of mucosa. Microscopically, epithelial abnormalities, inflammatory and neoplastic conditions were noticed. Epithelial abnormalities included epithelial hyperplasia, von Brunn’s nests, cystitis cystica and cystitis glandularis. Inflammatory conditions such as congestion, haemorrhages and cystitis were noticed. Cystitis was classified as lymphocytic, plasmo-lymphocytic, follicular and interstitial types. However, only single case of haemangiosarcoma was observed. Polymerase chain reaction revealed the presence of DNA of only BPV-1 in one sample out of 21, while one sample showed the presence of DNA of both BPV-1 and BPV-2. On immunohistochemistry, Ki-67 expression was observed only in haemangiosarcoma and one case of plasmo-lymphocytic cystitis with immunostaining restricted to nuclei. COX-2 immunostaining was observed in one case each of flat hyperplasia, von Brunn’s nest, haemangiosarcoma and two cases of plasmo-lymphocytic cystitis. Among epithelial abnormalities, COX-2 immunostaining was restricted to epithelial component of the lesions, and it was diffused cytoplasmic type with occasional perinuclear staining particularly in the cytoplasmic membrane. However, in haemangiosarcoma the COX-2 immunoreactivity was noticed in epithelial as well as connective tissue stroma. In conclusion, it may be stated that urinary bladders of slaughtered buffaloes showed a number of epithelial abnormalities, inflammatory conditions and occasionally neoplasms. Further studies to explore the infectious as well as non-infectious etiologies should be undertaken.

Palmatine, a natural alkaloid, attenuates memory deficits and neuroinflammation in mice submitted to permanent focal cerebral ischemia

Ischemic stroke is one of the major causes of human morbidity and mortality. The pathophysiology of ischemic stroke involves complex events, including oxidative stress and inflammation, that lead to neuronal loss and cognitive deficits. Palmatine (PAL) is a naturally occurring (Coptidis rhizome) isoquinoline alkaloid that belongs to the class of protoberberines and has a wide spectrum of pharmacological and biological effects. In the present study, we evaluated the impact of Palmatine on neuronal damage, memory deficits, and inflammatory response in mice submitted to permanent focal cerebral ischemia induced by middle cerebral artery (pMCAO) occlusion. The animals were treated with Palmatine (0.2, 2 and 20 mg/kg/day, orally) or vehicle (3% Tween + saline solution) 2 h after pMCAO once daily for 3 days. Cerebral ischemia was confirmed by evaluating the infarct area (TTC staining) and neurological deficit score 24 h after pMCAO. Treatment with palmatine (2 and 20 mg/kg) reduced infarct size and neurological deficits and prevented working and aversive memory deficits in ischemic mice. Palmatine, at a dose of 2 mg/kg, had a similar effect of reducing neuroinflammation 24 h after cerebral ischemia, decreasing TNF-, iNOS, COX-2, and NF- κB immunoreactivities and preventing the activation of microglia and astrocytes. Moreover, palmatine (2 mg/kg) reduced COX-2, iNOS, and IL-1β immunoreactivity 96 h after pMCAO. The neuroprotective properties of palmatine make it an excellent adjuvant treatment for strokes due to its inhibition of neuroinflammation.

Differential biochemical-inflammatory patterns in the astrocyte-neuron axis of the hippocampus and frontal cortex in Wistar rats with metabolic syndrome induced by high fat or carbohydrate diets

Metabolic syndrome (MetS) is a public health problem and a risk of developing cardiometabolic and neurodegenerative diseases. The biochemical-inflammatory impairment in brain areas related to learning and memory has not been differentiated between MetS models. We aimed to compare the effect of the MetS generated by consuming high-fat (HFD) or -carbohydrate diets (HCD) on the hippocampus and frontal cortex, related to astrocyte-neuron metabolism and neuroinflammation origin. Sixty male Wistar rats were separated into three groups: 1) control group, 2) HCD group, and 3) HFD group. After 3 months, we evaluated zoometry, a serum bioclinical profile, and in the hippocampus and frontal cortex, we performed biochemical assays (concentration of lactate, glutamate, fatty acids, and ASAT, ALAT, and LDH activity), immunoreactivity tests (GFAP, COX2, CD36, and BDNF), and immunoassays (TNF-α, IL-1β, IL-6, and PGE2). The bioclinical parameters showed that both diets induce MetS. At the brain level, it is noteworthy that the HCD group had an increase in lactate and glutamate concentration, reactive astrogliosis, immunoreactive COX2 neurons in the CA1 subfield hippocampus and frontal cortex, and high levels of PGE2, TNF-α, IL-1β, and IL-6, and low BDNF immunoreactivity. Meanwhile, the HFD is highlighted by increased fatty acid levels and CD36 expression in the hippocampus and frontal cortex, strong reactive astrogliosis and COX2 immunoreactivity, and the greatest inflammation with the lowest BDNF immunoreactivity. In conclusion, MetS induction by an HFD or HCD generates different biochemical, cellular, and inflammatory patterns in the hippocampus and frontal cortex.

Evaluation of the therapeutic role of allopurinol on bisphenol S gastric and renal toxicity in adult male albino rats: An in vivo study.

Bisphenol S (BPS) is used as an alternative to bisphenol A (BPA) in polycarbonate plastics, epoxy resins and thermal receipt sheet manufacturing. We examined the toxic effects of BPS on gastric and renal functions, as well as the efficacy of allopurinol as a treatment. Albino rats were given only BPS (30 and 120 mg/kg BW/day), and some were treated with allopurinol prior to sacrifice. Gastric and renal specimens were evaluated histologically and immunohistochemically, and blood from the tail vein was analysed for levels of gastrin, uric acid (UA), erythropoietin and 8-deoxyguanosine (8-OHdG). Gastrin levels decreased while erythropoietin, UA and 8-OHdG levels increased significantly. The severity of gastric and renal damage observed in BPS-treated animals increased with increasing doses. The mean percentage of COX-2 immunoreactivity and the mean number of CD45 immunoreactive cells were significantly increased in the stomach and kidney of BPS rats. Allopurinol ameliorated the biochemical, histological and immunohistochemical alterations induced by BPS, with superior protection at lower doses. Allopurinol can reverse the effects of BPS on the stomach and kidneys.

Piracetam attenuates cyclophosphamide-induced hepatotoxicity in rats: Amelioration of necroptosis, pyroptosis and caspase-dependent apoptosis

AimsCyclophosphamide (Cyclo) is an immunosuppressive and antineoplastic agent. The clinical use of Cyclo is limited by significant hepatotoxicity. Piracetam (Pira) is used to improve cognitive function. Pira possesses diverse physiological functions; however, the exact mechanisms of its activity are still non-elucidated. Main methodsForty rats were allocated in four groups. 1st group comprised normal rats; the remaining groups received single Cyclo dose (200 mg/kg/i.p.) on the experiment's 15th day. 2nd group comprised Cyclo-control rats. 3rd & 4th groups received Pira (100 & 300 mg/kg body weight) for 15 days. Key findingsCyclo administration resulted in deterioration of serum liver function tests and elevation of hepatic tissue concentration of P53, Nf-kβ, apoptosis-inducing factor-1, NLRP3 inflammasome, Bax; gene expression of receptor-induced protein-1 along with reduction of hepatic Bcl-2 concentration. Bax/Bcl-2 ratio headed for apoptosis. Cyclo administration also resulted in a severe deterioration of the hepatic histopathological picture and significant immunohistochemical expression of caspase-3, tumor necrosis factor-alpha (TNF-α) and Cyclooxygenase-2 (COX-2) in hepatic tissues versus the normal group. Pira significantly improved all the aforementioned parameters, reallocating the Bax/Bcl-2 ratio to anti-apoptosis. Moreover, Pira treatment amended Cyclo-induced histopathological abnormalities and significantly reduced caspase-3, TNF-α plus COX-2 immunoreactivity in hepatic tissues. SignificanceThe present work is the first to link Cyclo-induced hepatotoxicity to the activation of caspase-independent apoptosis (necroptosis), pyroptosis and caspase-dependent apoptosis signaling pathways. Pira treatment significantly ameliorated Cyclo-induced hepatotoxicity mainly via the amendment of necroptotic, pyroptotic and caspase-dependent apoptotic changes along with the histopathological deformities in rats' hepatic tissues.

Time dependent alteration of locomotor behavior in rat with acute liver failure induced cerebellar neuroinflammation and neuro-astroglial damage

Hepatic encephalopathy (HE) is a neurophysiological syndrome secondary to acute or chronic liver failure. Studies showed that HE patients exhibit a deficit in motor coordination, which may result from cerebellar functional impairment. The aim of this study is to assess the time-dependent alteration of locomotor behavior and the glial and neuronal alteration in rat with acute HE induced chemically. The study was carried out in male Sprague-Dawley rats with thioacetamide (TAA) induced acute liver failure at different stages 12 h, 24 h and 36 h. Hepatic and renal functions were assessed via various biochemical and histopathological examinations, while the cerebellum and the midbrain were examined using histology and immunohistochemistry for tyrosine hydroxylase (TH), cyclooxygenase-2 (COX-2) and glial fibrillary acidic protein (GFAP). We used as well, the open field test and the Rotarod test for assessing the locomotor activity and coordination. Our data showed a progressive loss of liver function and a progressive alteration in locomotor behavior and motor coordination in acute HE rats. In the cerebellum, we noted an increase in the degeneration of cerebellar Purkinje neurons parallel to increased COX-2 immunoreactivity together with astrocytic morphology and density changes. Likewise, in substantia nigra pars compacta, TH levels were reduced. We showed through the current study, a progressive deterioration in locomotor behavior in acute HE rats, as a result of Purkinje neurons death and a deficient dopaminergic neurotransmission, together with the morpho-functional astroglial modifications involving the oxidative stress and neuroinflammation.

Neuroprotective Effect of Fisetin Through Suppression of IL-1R/TLR Axis and Apoptosis in Pentylenetetrazole-Induced Kindling in Mice.

Epilepsy is a complex neurological disorder, characterized by frequent electrical activity in brain regions. Inflammation and apoptosis cascade activation are serious neurological sequelae during seizures. Fisetin (3, 3′,4′,7-tetrahydroxyflavone), a flavonoid molecule, is considered for its effective anti-inflammatory and anti-apoptotic properties. This study investigated the neuroprotective effect of fisetin on experimental epilepsy. For acute studies, increasing current electroshock (ICES) and pentylenetetrazole (PTZ)-induced seizure tests were performed to evaluate the antiseizure activity of fisetin. For the chronic study, the kindling model was established by the administration of PTZ in subconvulsive dose (25 mg/kg, i.p.). Mice were treated with fisetin (5, 10, and 20 mg/kg, p.o.) to study its probable antiseizure mechanism. The kindled mice were evaluated for seizure scores. Their hippocampus and cortex were assessed for neuronal damage, inflammation, and apoptosis. Histological alterations were observed in the hippocampus of the experimental mice. Levels of high mobility group box 1 (HMGB1), Toll-like receptor-4 (TLR-4), interleukin-1 receptor 1 (IL-1R1), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were assessed in the hippocampus and cortex by ELISA. The immunoreactivity and mRNA expressions of nuclear factor-κB (NF-κB), cyclooxygenase-2 (COX-2), cytochrome C, and caspase-3 were quantified by immunohistochemical analysis and real-time PCR. Phosphorylation ELISA was performed to evaluate AkT/mTOR (mammalian target of rapamycin) activation in the hippocampus and cortex of the kindled mice. The results showed that fisetin administration increased the seizure threshold current (STC) in the ICES test. In PTZ-induced seizures, fisetin administration increased the latency for myoclonic jerks (MJs) and generalized seizures (GSs). In the PTZ-induced kindling model, fisetin administration dose-dependently suppressed the development of kindling and the associated neuronal damage in the experimental mice. Further, fisetin administration ameliorated kindling-induced neuroinflammation as evident from decreased levels of HMGB1, TLR-4, IL-1R1, IL-1β, IL-6, and TNF-α in the hippocampus and cortex of the kindled mice. Also, the immunoreactivity and mRNA expressions of inflammatory molecules, NF-κB, and COX-2 were decreased with fisetin administration in the kindled animals. Decreased phosphorylation of the AkT/mTOR pathway was reported with fisetin administration in the hippocampus and cortex of the kindled mice. The immunoreactivity and mRNA expressions of apoptotic molecules, cytochrome C, and caspase-3 were attenuated upon fisetin administration. The findings suggest that fisetin shows a neuroprotective effect by suppressing the release of inflammatory and apoptosis molecules and attenuating histological alterations during experimental epilepsy.

Magnetic resonance imaging under isoflurane anesthesia alters cortical cyclooxygenase-2 expression and glial cell morphology during sepsis-associated neurological dysfunction in rats.

BackgroundMagnetic resonance imaging (MRI) of rodents combined with histology allows to determine what mechanisms underlie functional and structural brain changes during sepsis‐associated encephalopathy. However, the effects of MRI performed in isoflurane‐anesthetized rodents on modifications of the blood‐brain barrier and the production of vasoactive prostaglandins and glia cells, which have been proposed to mediate sepsis‐associated brain dysfunction, are unknown.MethodsThis study addressed the effect of MRI under isoflurane anesthesia on blood‐brain barrier integrity, cyclooxygenase‐2 expression, and glial cell activation during cecal ligature and puncture‐induced sepsis‐associated brain dysfunction in rats.ResultsCecal ligature and puncture reduced food intake and the righting reflex. MRI under isoflurane anesthesia reduced blood‐brain barrier breakdown, decreased circularity of white matter astrocytes, and increased neuronal cyclooxygenase‐2 immunoreactivity in the cortex 24 hours after laparotomy. In addition, it annihilated cecal ligature and puncture‐induced increased circularity of white matter microglia. MRI under isoflurane anesthesia, however, did not alter sepsis‐associated perivascular cyclooxygenase‐2 induction.ConclusionThese findings indicate that MRI under isoflurane anesthesia of rodents can modify neurovascular and glial responses and should, therefore, be interpreted with caution.

English

Cardiac arrest (CA) is a sudden interruption in the effective blood flow due to heart failure. The current research aimed to conduct the pathophysiological and histopathological analysis in the kidney in asphyxial cardiac arrest rat model. Cardiac arrest was induced by intravenous injection of vecuronium bromide (2 mg/kg), following stop of mechanical ventilation. Rats were kept on the CA condition for 5 minutes. After that, cardiopulmonary resuscitation (CPR) was done to achieve return of spontaneous circulation (ROSC) following intravenous injection of epinephrine bolus (0.005 mg/kg), sodium bicarbonate (1 mEq/kg) and turn on mechanical ventilation. Then Rats were sacrificed after cardiopulmonary resuscitation (CPR) following asphyxial CA at 6 hrs, 12 hrs, 1 day, 2 days, and 5 days. The intensity of renal injury measured by the serum levels of blood urea nitrogen (BUN), creatinine (Crtn). Moreover, Hematoxylin & eosin, and Periodic Acid Schiff staining in the kidney was done for evaluating the renal histopathological changes. Furthermore, COX-2 immunoreactivity and western analysis were performed in the kidney. Survival rate declined following ROSC compared to the sham group, it showed 80% at 6 hrs and decreased time-dependently to 8% at 5 days. In this study, serum BUN and Crtn levels and renal histopathological scores significantly increased after ROSC in CA. Moreover, COX-2 expression also increased after ROSC in comparison to the sham group with its peak level at 5 days following CA. Renal histological damage score and COX-2 expression were upregulated after ROSC following CA. These results direct that COX-2 takes part in the asphyxial CA-induced ischemic renal injury

Effects of vitamin B12 on methotrexate hepatotoxicity: evaluation of receptor-interacting protein (RIP) kinase.

In the study, we aimed to show the effects of vitamin B12 on the necrosis caused by methotrexate (MTX), a folic acid antagonist. Thirty-two rats were randomly assigned to four groups of eight rats per group. Control (n = 8), Vit B12 (n = 8) 3 μg/kg/ip B12 (15 days) per day throughout the experiment, MTX (n = 8) injected with a single dose of 20 mg/kg/ip MTX on 8th day of experiment, MTX + Vit B12 (n = 8) injected with a single dose of 20 mg/kg ip methotrexate on 8th day of experiment + 3 μg/kg/ip Vit B12 (15 days) per day throughout the experiment. Oxidant (TOS)/antioxidant (TAS) system, TNF-α and TGF-β levels, AST and ALT, serum vitamin B12 levels were determined in the tissue. Cyclooxygenase-2 (Cox-2), receptor-interacting protein kinase 1 (RIP1) and 3 (RIP3) immunohistochemistry were applied to the liver tissue. TOS increased; TAS decreased; TNF-α and TGF-β levels increased; AST and ALT levels changed after MTX hepatotoxicity. Vit B12 decreased significantly. COX-2, RIP1, and RIP3 immunoreactivity increased. Vit B12 showed improvement in all of the negative results. Vit B12 is an important supplement to be used against necrosis in tissue after MTX hepatotoxicity.

Metformin and silymarin afford protection in cyclosporine A induced hepatorenal toxicity in rat by modulating redox status and inflammation.

The use of cyclosporine A (CsA) as an immunosuppressive agent is often limited owing to its hepatotoxic and nephrotoxic properties. The present study was designed to evaluate the protective effect of metformin and silymarin in a rat model of CsA induced hepatorenal toxicity. The study included seven groups of Wistar albino rats (n = 6 per group): normal control, experimental control (CsA alone, 25 mg/kg), CsA + metformin (50 and 500 mg/kg), CsA + silymarin (50 and 200 mg/kg) and CsA + vitamin E (100 mg/kg). All the drugs were given daily for a period of 21 days by oral gavage and their effect was evaluated on serum levels of organ function markers (serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, bilirubin, urea/blood urea nitrogen, creatinine), markers of oxidative stress (thiobarbituric acid reactive substances, glutathione, superoxide dismutase), inflammation (nitrite, myeloperoxidase, tumour necrosis factor-alpha, prostaglandin E2 ), apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labelling positivity) in addition to tissue histology, cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) immunoreactivity. Administration of metformin and silymarin along with CsA ameliorated functional damage to liver and kidneys in a dose-dependent manner. Significant and comparable improvement in the tissue levels of oxidative stress, inflammation, apoptotic markers was also observed following treatment with both the test drugs. Normalization of histology scores, as well as COX-2 and iNOS immunoreactivity scores, further strengthened these findings. The hepatoprotective and nephroprotective effects of metformin and silymarin were comparable and matched with that of reference drug, vitamin E. The findings of the present study suggest that both metformin and silymarin have a potential for clinical use in patients receiving long-term CsA treatment to maintain their liver and kidney functions.

A Comparative study on the protective role of Silymarin and Coenzyme-Q10 on the cerebellar cortex of experimentally induced atherosclerosis in adult male albino rats: A Histological, Immunohistochemical and Biochemical study

Background: Atherosclerosis is known as a serious cardiovascular condition that causes human mortality worldwide. Atherosclerosis is a significant progressive condition of inflammation. Atherosclerosis of the cerebellar arteries affects voice, balance and fine motor control. Coenzyme-Q10 vitamin soluble in fat that used for heart failure and angina treatment protocol. It has the potential to influence cellular bioenergetics, and counteract free radical damage. This has proven success in treating many neurodegenerative diseases. Silymarin is a flavonoid extract of the herb Silybum marianum. It has been reported as the gold medicine in liver disorders and evaluated recently as a neuroprotective.Objective: Comparison of potential neuroprotective effects of Co Q10 and Silymarin on adult male albino rats cerebellar cortex suffering from atherosclerosis.Materials and Methods: Sixty adult male albino rats were divided randomly into equal four classes: Group I (control group) subdivided into three subgroups: Subgroup IA: provided ordinary diet. Subgroup IB (Co enzyme Q10 treated): obtained ordinary diet and co-enzyme Q10 Subgroup IC (Silymarin treated): obtained ordinary diet and Silymarin. Group II (atherosclerotic group), Group III (atherosclerotic Co Q10 treated group) and Group IV (atherosclerotic Silymarin treated group). After three months, lipid profile was assessed in blood samples taken from all classes. Cerebellar cortices were processed for histological, biochemical, immunohistochemical and electron microscopic studies.Results: Atherosclerotic rats showed loss and disruption of Purkinje cell layer with pyknotic nuclei, dilated cisternae of rough endoplasmic reticulum, prominent perineural spaces and spongiosis of granular and molecular layers. Significant increase in GFAP, Cox-2 immunoreactivity was also found. However, concomitant administration of Co Q10 or Silymarin had ameliorating effect on these changes.Conclusion: Co Q10 or Silymarin supplementation had a neurological protective role in atherosclerosis. Silymarin may be the ideal neuroprotective agent and new approach for reduction of many neurological diseases in high risk population.

Immunohistochemical Expression of Fibroblast Growth Factor Receptor 3 and Cyclooxygenase-2 in Urinary Bladder Carcinomas with Correlation of Schistosomiasis in Egyptian Patients

BACKGROUND: In Egypt, schistosomal infestation is a leading cause of bladder cancer. A mutation in the fibroblast growth factor receptor 3 (FGFR-3) gene is the most common and most specific genetic abnormality in bladder cancer. Similarly, cyclooxygenase-2 (COX-2) is an inducible, pro-inflammatory enzyme with previous studies showing higher expression in schistosomal-associated bladder cancer.
 AIM OF THE STUDY: The aim of the study was to evaluate the immunohistochemical expression of FGFR3 and COX2 in bladder carcinoma and correlates their expression to the associated schistosomal infestation to implicate possible therapeutic treatments.
 MATERIALS AND METHODS: This retrospective study included a total of 90 cases of archived, formalin fixed, paraffin-embedded tissue blocks that included variable subtypes and grades of urothelial carcinomas. Immunohistochemistry for expression of FGFR-3 and COX2 was performed using a standard avidin-biotin-peroxidase system.
 RESULTS: About 73.3% of the total cases (66 cases) showed variable positive reactivity for FGFR3, of which 33.3% (22 cases) were associated with bilharzia infection. A statistically significant correlation was detected between FGFR-3 and tumor size, grade, histologic subtype, LN status, lymphovascular invasion, and stage. About 83.3% of the total cases (75 cases) showed variable positive immunoreactivity for COX-2, of which 37.3% (28 cases) were bilharzial-associated. A positive correlation was established between COX-2 and grade, concomitant in situ changes and cases associated with bilharzia infection.
 CONCLUSION: FGFR-3 can be used as a prognostic marker for low-grade urothelial tumors. Results also portray that COX-2 has an inflammatory inciting role in bladder carcinoma development, especially in patients with a history of schistosomiasis (bilharziasis). Both COX-2 and FGFR-3 should be explored further for its use alone or in combination with conventional treatment, to reduce the recurrence rate and progression of superficial (low grade) tumors.

Peripheral-to-central immune communication at the area postrema glial-barrier following bleomycin-induced sterile lung injury in adult rats

The pathways for peripheral-to-central immune communication (P → C I-comm) following sterile lung injury (SLI) are unknown. SLI evokes systemic and central inflammation, which alters central respiratory control and viscerosensory transmission in the nucleus tractus solitarii (nTS). These functional changes coincide with increased interleukin-1 beta (IL-1β) in the area postrema, a sensory circumventricular organ that connects P → C I-comm to brainstem circuits that control homeostasis. We hypothesize that IL-1β and its downstream transcriptional target, cyclooxygenase-2 (COX-2), mediate P → C I-comm in the nTS. In a rodent model of SLI induced by intratracheal bleomycin (Bleo), the sigh frequency and duration of post-sigh apnea increased in Bleo- compared to saline- treated rats one week after injury. This SLI-dependent change in respiratory control occurred concurrently with augmented IL-1β and COX-2 immunoreactivity (IR) in the funiculus separans (FS), a barrier between the AP and the brainstem. At this barrier, increases in IL-1β and COX-2 IR were confined to processes that stained for glial fibrillary acidic protein (GFAP) and that projected basolaterally to the nTS. Further, FS radial-glia did not express TNF-α or IL-6 following SLI. To test our hypothesis, we blocked central COX-1/2 activity by intracerebroventricular (ICV) infusion of Indomethacin (Ind). Continuous ICV Ind treatment prevented Bleo-dependent increases in GFAP + and IL-1β + IR, and restored characteristics of sighs that reset the rhythm. These data indicate that changes in sighs following SLI depend partially on activation of a central COX-dependent P → C I-comm via radial-glia of the FS.

Targeting Cyclooxygenase-2 in Pheochromocytoma and Paraganglioma: Focus on Genetic Background.

Cyclooxygenase 2 (COX-2) is a key enzyme of the tumorigenesis-inflammation interface and can be induced by hypoxia. A pseudohypoxic transcriptional signature characterizes pheochromocytomas and paragangliomas (PPGLs) of the cluster I, mainly represented by tumors with mutations in von Hippel–Lindau (VHL), endothelial PAS domain-containing protein 1 (EPAS1), or succinate dehydrogenase (SDH) subunit genes. The aim of this study was to investigate a possible association between underlying tumor driver mutations and COX-2 in PPGLs. COX-2 gene expression and immunoreactivity were examined in clinical specimens with documented mutations, as well as in spheroids and allografts derived from mouse pheochromocytoma (MPC) cells. COX-2 in vivo imaging was performed in allograft mice. We observed significantly higher COX-2 expression in cluster I, especially in VHL-mutant PPGLs, however, no specific association between COX-2 mRNA levels and a hypoxia-related transcriptional signature was found. COX-2 immunoreactivity was present in about 60% of clinical specimens as well as in MPC spheroids and allografts. A selective COX-2 tracer specifically accumulated in MPC allografts. This study demonstrates that, although pseudohypoxia is not the major determinant for high COX-2 levels in PPGLs, COX-2 is a relevant molecular target. This potentially allows for employing selective COX-2 inhibitors as targeted chemotherapeutic agents and radiosensitizers. Moreover, available models are suitable for preclinical testing of these treatments.