Amelioration of functional,biochemical and molecular deficits by epigallocatechin gallate in experimental model of alcoholic neuropathy

Long term alcohol consumption leads to decreased nociceptive threshold characterized by spontaneous burning pain, hyperalgesia and allodynia. The mechanism involved in this pain includes increased oxidative‐nitrosative stress, release of pro‐inflammatory cytokines and neuronal apoptosis. The present study was designed to explore the protective effect of epigallocatechin‐3‐gallate against alcoholic neuropathic pain in rats. Rats fed with alcohol (35%) for 10 weeks showed markedly decreased tail flick latency in tail‐immersion test (thermal hyperalgesia), vocalization threshold in Randall—Sellito test (mechanical hyperalgesia) and paw‐withdrawal threshold in von‐Frey hair test (mechanical allodynia) along with enhanced oxidative‐nitrosative stress and inflammatory mediators (TNF‐α, IL‐1β and TGF‐β1 levels). Co‐administration of epigallocatechin‐3‐gallate (25–100 mg/kg) significantly and dose‐dependently prevented functional, biochemical and molecular changes associated with alcoholic neuropathy. In conclusion, the current findings suggest the neuroprotective potential of epigallocatechin‐3‐gallate in attenuating the functional, biochemical and molecular alterations associated with alcoholic neuropathy through modulation of oxido‐inflammatory cascade.     

Comparative study of alpha‐ and beta‐pinene effect on PTZ‐induced convulsions in mice

Convulsions occur in response to a loss of balance between excitatory and inhibitory neurotransmitters, and the treatment for this condition consists in restore such lost balance. Many anticonvulsant drugs present side effects which may limit their use. This fact has stimulated the search for new sources of treatment from aromatic plants. Many monoterpenes commonly present in essential oils are known because of their anticonvulsant properties. The anticonvulsant effect of α‐ and β‐pinene, two structural isomers, is still little studied. Thus, the present work evaluated the anticonvulsant effect of α‐ and β‐pinene in pentylenetetrazole‐induced convulsions model. Initially, the oral LD₅₀ for α‐ and β‐pinene was estimated. Following the oral administration, a mild sedation was observed and no deaths were recorded; the LD₅₀ estimated for both monoterpenes was greater than 2 000 mg/kg, p.o. Further, animals were orally treated with α‐pinene (100, 200 and 400 mg/kg), β‐pinene (100, 200 and 400 mg/kg) and the equimolar mixture of α‐ and β‐pinene (400 mg/kg) and subjected to the pentylenetetrazole‐induced convulsions model. In this model, only the dose of 400 mg/kg of the compounds was able to significantly decrease the seizure intensity. The latency of first convulsion was significantly increased by the mixture of α‐ and β‐pinene (400 mg/kg). In addition, β‐pinene and the mixture of the two monoterpenes, both at a dose of 400 mg/kg, significantly increased the time of death of animals. The treatment with β‐pinene and the equimolar mixture of the two monoterpenes significantly reduced hippocampal nitrite level and striatal content of dopamine (DA) and norepinephrine (NE). Taken together, the results suggest that α‐pinene appears to be devoid of anticonvulsant action. This fact, however, seems to be dependent on the chemical structure of the compound, since pretreatment with the β‐pinene increased the time of death pf PTZ‐treated mice, which seems to depend on the ability of the compound to reduce nitrite concentration and NE and DA content, during the pentylenetetrazole‐induced seizure.     

Tocotrienol ameliorates behavioral and biochemical alterations in the rat model of alcoholic neuropathy

Chronic alcohol consumption produces a painful peripheral neuropathy for which there is no reliable successful therapy, which is mainly due to lack of understanding of its pathobiology. Alcoholic neuropathy is characterized by spontaneous burning pain, hyperalgesia (an exaggerated pain in response to painful stimuli) and allodynia (a pain evoked by normally innocuous stimuli). Chronic alcohol intake is known to decrease the nociceptive threshold with increased oxidative–nitrosative stress and release of proinflammatory cytokines coupled with activation of protein kinase C. The aim of the present study is to investigate the effect of both isoforms of vitamin E, α-tocopherol (100 mg/kg; oral gavage) and tocotrienol (50, 100 and 200 mg/kg; oral gavage) against alcohol-induced neuropathic pain in rats. Male Wistar rats, were administered 35% v/v ethanol (10 g/kg; oral gavage) for 10 weeks, and were treated with α-tocopherol and tocotrienol for the same duration. Ethanol-treated animals showed a significant decrease in nociceptive threshold as evident from decreased tail flick latency (thermal hyperalgesia) and decreased paw-withdrawal threshold in Randall–Sellito test (mechanical hyperalgesia) and von-Frey hair test (mechanical allodynia) along with the reduction in nerve glutathione and superoxide dismutase levels. TNF-α and IL-1β levels were also significantly increased in both serum and sciatic nerve of ethanol-treated rats. Treatment with α-tocopherol and tocotrienol for 10 weeks significantly improved all the above-stated functional and biochemical deficits in a dose-dependent manner with more potent effects observed with tocotrienol. The study demonstrates the effectiveness of tocotrienol in attenuation of alcoholic neuropathy.     

Comparative study between atorvastatin and losartan on high fat diet‐induced type 2 diabetes mellitus in rats

Obesity is often associated with chronic inflammatory state which contributes to the development of insulin resistance (IR) and type 2 diabetes mellitus (T2DM). This study investigated the effects of single and combined administration of atorvastatin (ATOR, lipid‐lowering drug) and losartan (LOS, angiotensin receptor antagonist) on metabolic disorders and inflammatory status that are implicated in the development of T2DM with the use of pioglitazone (PIO) as a standard antidiabetic drug. T2DM was induced in male rats by high‐fat diet (HFD) feeding for 16 weeks. Oral administrations of ATOR (10 mg/kg), LOS (20 mg/kg), PIO (3 mg/kg), their binary combinations, or vehicle were started in the last 4 weeks. Fasting serum glucose, oral glucose tolerance, fasting serum insulin, IR, serum lipid profile, serum TNF‐α and body composition index were determined. Results showed that all drugs and their combinations had positive impact effect on all measured parameters, and better results were achieved from binary drug combinations than administration of each drug alone. Combination of PIO with either ATOR or LOS provided better improvements on T2DM‐associated metabolic abnormalities and inflammatory status with respect to each drug alone. However, the most pronounced effects of drugs and their combinations regarding the above parameters were attributed to LOS + PIO combination. In conclusion, this study indicates that combination of ATOR + PIO and, in particular, LOS + PIO can be used as promising effective therapies in the management of HFD‐induced T2DM. This concept may be attributed to the combined effects of the respective monotherapies to improve lipid profile, insulin sensitivity, and TNF‐α level.     

Anti‐inflammatory effect of 4‐methylcyclopentadecanone in rats submitted to ischemic stroke

This study aimed to investigate the anti‐inflammatory effect of 4‐methylcyclopentadecanone (4‐MCPC) in rats suffering from a cerebral ischemia/reperfusion (I/R) injury. In this study, the focal cerebral ischemia in rats was induced by middle cerebral artery occlusion (MCAO) for 2 h, and the rats were treated with 4‐MCPC (8 mg/kg) just 0.5 h before reperfusion. The ischemic infarct volume was recorded 24 h after the MCAO. In addition, myeloperoxidase (MPO) activity and TNF‐α and IL‐1β levels in the ischemic cerebral cortex were determined by ELISA, while nuclear translocation of NF‐κB p65 subunit and expression of p‐IκBα were investigated by Western blotting. Our results showed that 4‐MCPC treatment decreased infarct volume significantly, compared with I/R group (16.8%±7.5% vs. 39.7%±10.9%); it reduced MPO activity (0.43 ± 0.10 vs. 1.00 ± 0.51 U/g) and expression levels of TNF‐α (18.90 ± 3.65 vs. 35.87 ± 4.87 ng/g) and IL‐1β (1.68 ± 0.23 vs. 2.67 ± 0.38 ng/g) in ischemic brain tissues of rats. Further study revealed that 4‐MCPC treatment markedly reduced nuclear translocation of NF‐κB p65 subunit and expression of p‐IκBα in ischemic cerebral cortex. Taken together, our results suggest that 4‐MCPC protects against cerebral I/R injury and displays anti‐inflammatory actions through inhibition of the NF‐κB signal pathway.     

Sumatriptan reduces severity of status epilepticus induced by lithium–pilocarpine through nitrergic transmission and 5‐HT1B/D receptors in rats: A pharmacological‐based evidence

Status epilepticus (SE) is a life‐threatening neurologic disorder that can be as both cause and consequence of neuroinflammation. In addition to previous reports on anti‐inflammatory property of the anti‐migraine medication sumatriptan, we have recently shown its anticonvulsive effects on pentylenetetrazole‐induced seizure in mice. In the present study, we investigated further (i) the effects of sumatriptan in the lithium–pilocarpine SE model in rats, and (ii) the possible involvement of nitric oxide (NO), 5‐hydroxytryptamin 1B/1D (5‐HT_1B/1D) receptor, and inflammatory pathways in such effects of sumatriptan. Status epilepticus was induced by lithium chloride (127 mg/kg, i.p) and pilocarpine (60 mg/kg, i.p.) in Wistar rats. While SE induction increased SE scores and mortality rate, sumatriptan (0.001‐1 mg/kg, i.p.) improved it (P < 0.001). Administration of the selective 5‐HT_1B/1D antagonist GR‐127935 (0.01 mg/kg, i.p.) reversed the anticonvulsive effects of sumatriptan (0.01 mg/kg, i.p.). Although both tumor necrosis factor‐α (TNF‐α) and NO levels were markedly elevated in the rats' brain tissues post‐SE induction, pre‐treatment with sumatriptan significantly reduced both TNF‐α (P < 0.05) and NO (P < 0.001) levels. Combined GR‐127935 and sumatriptan treatment inhibited these anti‐inflammatory effects of sumatriptan, whereas combined non‐specific NOS (L‐NAME) or selective neuronal NOS (7‐nitroindazole) inhibitors and sumatriptan further reduced NO levels. In conclusion, sumatriptan exerted a protective effect against the clinical manifestations and mortality rate of SE in rats which is possibly through targeting 5‐HT_1B/1D receptors, neuroinflammation, and nitrergic transmission.     

Modulation of heme oxygenase‐1 expression and activity affects streptozotocin‐induced diabetic nephropathy in rats

Heme oxygenase (HO)‐1 has exhibited nephro‐protective actions in different animal models; however, its full mechanistic potential in diabetic nephropathy (DN) has not yet been elucidated. Hence, the present study has been undertaken by inducing DN in rats using streptozotocin (50 mg/kg i.p.), with or without either HO‐1 inducer; hemin (HM; 40 μmol/kg, s.c.), or HO‐1 blocker; zinc protoporphyrin‐IX (ZnPP; 50 μmol/kg, i.p.), for one month. Compared to control, rats with DN suffered from hyperglycemia and hyperlipidemia, with signs of renal damage, as assessed by distortion in renal histopathologic architecture and kidney function. Renal oxidative/nitrosative stress was evident by increased malondialdehyde, nitric oxide, myeloperoxidase, with decreased reduced glutathione, superoxide dismutase, and catalase. DN group also exhibited high renal expression of the pro‐inflammatory cytokine; tumor necrosis factor (TNF)‐α, and the apoptotic marker; caspase 3, assessed by Western blot. Renal HO‐1 protein expression and activity were increased in DN rats compared to control. Administration of HM, but not ZnPP, to DN rats improved kidney function, histopathologic features, lipid profile, TNF‐α, and caspase 3 expressions, with no effect on blood glucose level. HM increased, while ZnPP decreased renal HO‐1 activity in DN rats. It is noteworthy that neither intervention affected HO‐1 activity or renal oxidative capacity in non‐diabetic rats. Interestingly, the expression of HO‐1 was upregulated by both HM and ZnPP in DN rats. In conclusion, activation of HO‐1 via HM ameliorated renal damage in STZ‐induced DN in rats, probably through antioxidant, anti‐nitrosative, anti‐inflammatory, and anti‐apoptotic mechanisms.     

Quercetin reverses experimental cirrhosis by immunomodulation of the proinflammatory and profibrotic processes

The ability of quercetin to reverse an established cirrhosis has not yet been investigated. Therefore, the aim of this study was to examine the efficacy of this flavonoid in reversing experimental cirrhosis. Cirrhosis was induced by intraperitoneal administration of TAA (200 mg/kg of body weight) three times per week for 8 weeks or by intraperitoneal petrolatum‐CCl₄ (400 mg/kg of body weight) administration three times per week for 8 weeks. To determine the capacity of quercetin to prevent liver fibrosis, the flavonoid (50 mg/kg of body weight, p.o.) was administered daily to rats during the CCl₄ or TAA treatment. To evaluate the ability of quercetin to reverse the previously induced cirrhosis, we first treated rats with CCl₄ for 8 weeks, as previously described and then the flavonoid was administered for four more weeks. We found that the liver anti‐inflammatory and antinecrotic effects of quercetin are associated with its antioxidant properties, to the ability of the flavonoid to block NF‐κB activation and in consequence to reduce cytokine IL‐1. The ability of quercetin to reverse fibrosis may be associated with the capacity of the flavonoid to decrease TGF‐β levels, hepatic stellate cell activation, and to promote degradation of the ECM by increasing metalloproteinases. The main conclusion is that quercetin, in addition to its liver protective activity against TAA chronic intoxication, is also capable of reversing a well‐stablished cirrhosis by blocking the prooxidant processes and by downregulating the inflammatory and profibrotic responses.     

The role of interleukin‐1b and its antagonist (diacerein) in estradiol benzoate‐induced endometrial hyperplasia and atypia in female rats

Endometrial hyperplasia (EH) is a common gynecological condition and may progress to carcinoma. We investigated the effect of diacerein (DIA) on estradiol benzoate (EB)‐induced EH and atypia. DIA (50 mg/kg/day) was administered orally to rats for 4 weeks, in the presence or absence of EH induced by intramuscular injection of EB (60 μg/100 g) three times per week for 4 weeks. We measured levels of serum total cholesterol, uterine tissue malondialdehyde (MDA), total nitrites (NO_x), superoxide dismutase (SOD) activity, caspase‐3, interleukin‐1b (IL‐1b) immunoexpressions, and histopathology. Results showed that EB‐induced EH and atypia manifested by significant increase in serum total cholesterol with increase in MDA and NO_x levels. EB showed the typical histopathological changes of EH and atypia. In addition, there was reduction in SOD activity and decrease in caspase‐3 immunoexpressions but increase in IL‐1b immunoexpressions. DIA was able to reduce EB‐induced pathological changes.     

Branched chain amino acids supplementation modulates TGF‐β1/Smad signaling pathway and interleukins in CCl4‐induced liver fibrosis

The alterations and low levels of circulating branched chain amino acids (BCAAs), leucine, isoleucine, and valine, are associated with liver diseases. The study was designed to evaluate hepatoprotective effect of BCAAs on CCl₄‐induced liver fibrosis and to investigate the molecular mechanisms underlying these effects in rats. In all, 30 male rats were divided into three groups. Control group (n = 10) and CCl₄ group (n = 10), where rats were injected with CCl₄ (1 mL/kg of 0.5 : 1 v/v injected i.p. twice weekly for 12 weeks). In CCl₄ + BCAAs group (n = 10), rats were injected with similar doses of CCl₄ and supplemented with a mixture of 600 mg/kg BCAAs (2 : 1 : 1.2 leucine : isoleucine : valine) by oral gavage, three times/week for 12 weeks. Liver fibrosis was assessed by measuring total bilirubin, total protein, alanine aminotransferase, and aspartate aminotransferase, hydroxyproline content, and serum IL‐6 and IL‐10. Histopathologic studies and α‐smooth muscle actin (α‐SMA) were detected immunohistochemically in liver. Serum insulin level, blood glucose, liver malodialdehyde concentration (MDA), glutathione peroxidase, and superoxide dismutase (SOD) activities were quantified. TGF‐β1, Smad3, and Smad7 gene expressions were estimated by qRT‐PCR. BCAAs suppressed liver fibrosis induced by CCl₄ treatment. BCAAs modulated liver indices and downregulated TGF‐β1, Smad3, and Smad7 expressions in hepatocytes. BCAAs enhanced liver antioxidant enzyme activities (P < 0.001), reduced serum levels of TGF‐β1, IL‐6, and IL‐10 compared to CCL₄ group and ameliorated histopathologic changes in rat liver. BCAAs may have a protective role against liver fibrosis via antioxidant and anti‐inflammatory mechanisms.     

Neuropharmacological effects of lipoic acid and ubiquinone on the mRNA level of interleukin‐1β and acetylcholinesterase activity in rat hippocampus after seizures

The purpose of this study was to investigate the neuroprotective effects of lipoic acid and ubiquinone on interleukin‐1β (IL‐1β) mRNA levels and acetylcholinesterase (AChE) activities in rat hippocampus after pilocarpine‐induced seizures. Wistar rats were intraperitoneally administered with either 0.9% saline (icontrol group), LA (10 or 20 mg/kg, LA10 or LA20 groups), UQ (20 or 40 mg/kg, UQ20 and UQ40 groups), pilocarpine (400 mg/kg, P400 group), or co‐administration of pilocarpine with LA or UQ groups 30 min prior to LA or UQ administration. After the treatments, all groups were observed for 1 h. IL‐1β mRNA and AChE activity in rat hippocampus at 1 h after SE onset was determined. Results showed that rats pretreated with LA or UQ developed less seizures and SE more slowly and has less number than animals treated with pilocarpine alone. Reduced IL‐1β mRNA and marked AChE activities in the hippocampus were significantly higher in rats pretreated with LA or UQ in comparison with the values of the control and seized groups. Our findings strongly support the hypothesis that an increase on IL‐1β mRNA levels in hippocampus occurs during seizures induced by pilocarpine, which indicates that inflammatory process plays a crucial role in seizures pathogenic consequences. Our result also suggests that LA or UQ can exert significant neuroprotective effects, at least in part, because of the increase in the AChE activities in rat hippocampus that will be useful in the treatment of neurodegenerative diseases.     

α-Glucosidase inhibitors in diabetes: lessons from animal studies

Two rat models for non-insulin-dependent diabetes mellitus (NIDDM) have been used in our laboratory to study the effects of α-glucosidase inhibitors. These models become hyperglycaemic and have other characteristics which make them good models for NIDDM, and both prevention and reversal studies have been carried out; the prevention experiments were started before the animal became diabetic while the reversal groups were treated after diabetes had fully developed. In both models blood glucose was significantly lowered toward control levels using a dose of 40 mg per 100 g of diet while there was a less dramatic, but still significant, correction with half that dose. Treatment increased the weight gain of the more diabetic model (ZDF) while there was no effect of treatment on the weight of the Wistar diabetic fatty (WDF) rat. Other parameters such as glycated haemoglobins, nerve conduction velocity and nerve sugar content are also reversed with effective treatment of the hyperglycaemic condition.     

Neuropharmacological effects of lipoic acid and ubiquinone on δ‐aminolevulinic dehydratase,Na+, K+‐ATPase,and Mg2+‐ATPase activities in rat hippocampus after pilocarpine‐induced seizures

In this study, we investigated the effects of lipoic acid (LA) in the hippocampus oxidative stress caused by pilocarpine‐induced seizures in adult rats. Wistar rats were treated with 0.9% saline (i.p., control group), LA (10 mg/kg, i.p., LA group), ubiquinone [20 mg/kg, i.p., ubiquinone (UQ) group], pilocarpine (400 mg/kg, i.p., P400 group), and the association of LA (10 mg/kg, i.p.) plus pilocarpine (400 mg/kg, i.p.) or UQ (20 mg/kg, i.p.) plus pilocarpine (400 mg/kg, i.p.), 30 min before of administration of P400 (LA plus P400 group and UQ plus P400 group, respectively). After the treatments, all groups were observed for 1 h. The enzyme activities (δ‐aminolevulinic dehydratase (δ‐ALA‐D), Mg²⁺‐ATPase, and Na⁺, K⁺‐ATPase) were measured using spectrophotometric methods, and the results compared to values obtained from saline and pilocarpine‐treated animals. Protective effects of LA and UQ were also evaluated on the same parameters. We reported here for the first time that Na⁺, K⁺‐ATPase and δ‐ALA‐D activities inhibition and Mg²⁺‐ATPase stimulation in the pilocarpine model are probably attributed to the oxidative stress caused by seizures in the rat hippocampus. The addition of the antioxidants LA and UQ may reverses the previously mentioned Na⁺, K⁺‐ATPase and δ‐ALA‐D inhibitions and Mg²⁺‐ATPase stimulation. Conclusions: The oxidative stress plays an important signaling role in pilocarpine‐induced seizures, and antioxidant drugs might be considered as therapeutical tools in this pathology.     

Tauroursodeoxycholic acid produces antidepressant‐like effects in a chronic unpredictable stress model of depression via attenuation of neuroinflammation,oxido‐nitrosative stress,and endoplasmic reticulum stress

Depression is a common psychiatric disorder with heavy economic and social burdens. Searching new agents with better antidepressant‐like activities is of great significance for depression therapy. Tauroursodeoxycholic acid (TUDCA), a clinical drug for gallstone treatment, possesses neuroprotective effects in different brain disorders. However, whether it affects depression remains unclear. We addressed this issue by evaluating the effect of TUDCA on depression induced by chronic unpredictable stress (CUS). Results showed that TUDCA treatment at 200 but not 100 mg/kg prevented the 5 weeks of CUS‐induced increases in the immobile time of C57BL6/J mice in the experiments of forced swimming test and tail suspension test as well as the CUS‐induced decrease in sucrose intake and crossing numbers in the open‐field test, and did not enhance the antidepressant‐like effect of fluoxetine. Attenuation of neuroinflammation may be involved in the antidepressant‐like effect of TUDCA, as TUDCA treatment (200 mg/kg) normalized the levels of tumor necrosis factor‐α and interleukin‐6 in both hippocampus and prefrontal cortex. The increases in inflammasome and microglial activation markers, including interleukin‐β, nod‐like receptor protein 3, and Iba‐1, in CUS‐treated mice were reduced by TUDCA treatment (200 mg/kg). TUDCA treatment (200 mg/kg) also normalized the changes in markers reflecting the oxidative–nitrosative and endoplasmic reticulum (ER) stress induced by CUS, such as nitric oxide, reduced glutathione, malondialdehyde, glucose‐regulated protein 78, and C/EBP homologous protein. These results revealed that TUDCA improved the CUS‐induced depression‐like behaviors likely through attenuation of neuroinflammation, oxido‐nitrosative, and ER stress.     

Agmatine co‐treatment attenuates allodynia and structural abnormalities in cisplatin‐induced neuropathy in rats

Cisplatin is a widely used antineoplastic agent in the treatment of various cancers. Peripheral neuropathy is a well‐known side effect of cisplatin and has potential to result in limiting and/or reducing the dose, decreasing the quality of life. Thus, effective treatments are needed. Agmatine is an endogenous neuromodulator that has been shown to exert antiallodynic effects in various animal studies. The first aim of this study was to investigate the in vitro effects of agmatine on cisplatin‐induced neurotoxicity. Primary cultures of dorsal root ganglia (DRG) which are the primary target of drug injury were prepared. DRG cells were incubated with cisplatin (100, 200, 500 μm ). Then, agmatine (10, 100, 500 μm ) was administered with the submaximal concentration of cisplatin. Cisplatin caused concentration‐dependent neurotoxicity, and agmatine did not alter this effect. The second aim was to investigate the effects of agmatine on cisplatin‐induced peripheral neuropathy in rats and the influence of nitric oxide synthase (NOS) inhibitor, L‐NAME, in this effect. Female Sprague Dawley rats received intraperitoneal saline (control), cisplatin (3 mg/kg), cisplatin+agmatine (100 mg/kg), or cisplatin+agmatine+L‐NAME (10 mg/kg) once a week for 5 weeks. The mechanical allodynia, hot plate, and tail clip tests were performed, and DRG cells and sciatic nerves were analyzed. Agmatine and agmatine+L‐NAME combination attenuated CIS‐induced mechanical allodynia and degeneration in DRG cells and sciatic nerves. However, L‐NAME did not potentiate the antiallodynic or neuroprotective effect of agmatine. These findings indicate that agmatine co‐administration ameliorates cisplatin‐induced neuropathy and may be a therapeutic alternative.     

Potential of telmisartan in the treatment of benign prostatic hyperplasia

Benign prostatic hyperplasia (BPH) is a common health problem in ageing men. This study was carried out to investigate the protective effect of telmisartan on testosterone‐induced BPH in rats. Fifty‐four male Wistar rats (200–250 g) were randomly divided into nine groups (n = 6) and orally treated for 28 consecutive days: group 1 – vehicle normal, olive oil (10 mL/kg); group 2 – BPH model control (10 mL/kg); groups 3–5 – telmisartan (5, 10 or 20 mg/kg, respectively); group 6 – pioglitazone (20 mg/kg); group 7 – celecoxib (20 mg/kg); group 8 – combination of telmisartan (5 mg/kg) and pioglitazone (20 mg/kg); group 9 – combination of telmisartan (5 mg/kg) and celecoxib (20 mg/kg). Animals in groups 2–9 were given testosterone propionate in olive oil (3 mg/kg) subcutaneously 15 min after pretreatments. On day 29, blood was collected for the estimation of serum testosterone and prostate‐specific antigen (PSA). The prostates were excised, weighed and subjected to biochemical and histological studies. Testosterone injection induced significant increase in prostatic index, serum testosterone and PSA suggesting BPH as well as increased prostate oxidative stress which were ameliorated with the pretreatment of rats with telmisartan or co‐administration of celecoxib and pioglitazone. Histological examination showed that testosterone disrupted the morphology of the prostate epithelial cells evidenced in the involution of the epithelial lining of the acini into the lumen indicating BPH which was reversed by telmisartan. Findings from this study showed that telmisartan alone or in combination with pioglitazone prevented the development of testosterone‐induced prostatic hyperplasia.     

Protective effect of (−)-epigallocatechin-gallate (EGCG) on lipid peroxide metabolism in isoproterenol induced myocardial infarction in male Wistar rats: A histopathological study

This study aims to evaluate the preventive effect of (−)-epigallocatechin-gallate (EGCG) on lipid peroxides, enzymatic and non-enzymatic antioxidants and histopathological findings in isoproterenol (ISO)-induced rats. Myocardial infarction (MI) is induced in rats by subcutaneous injection of ISO (100 mg/kg body weight) at an interval of 24 h for 2 days. ISO-treated rats show a significant increase in the levels of thiobarbituric acid reactive substances, lipid hydroperoxides in plasma and heart and plasma uric acid and a significant decrease in the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase in heart and the levels of reduced glutathione, vitamin C and vitamin E in plasma and the heart and ceruloplasmin in plasma. Oral pretreatment with EGCG (10, 20 and 30 mg/kg body weight) daily for a period of 21 days show significant decrease in the levels of lipid peroxidation products and uric acid and improved the antioxidant status by increasing the activities of antioxidant enzymes and non-enzymic antioxidants. Histopathological findings of the myocardial tissue show the protective effect of EGCG in ISO-induced rats. The effect at a dose of 30 mg/kg of EGCG was more pronounced than that of the other two doses (10 and 20 mg/kg body weight). Thus, the present study reveals that EGCG exerts cardioprotective effect against ISO-induced MI due to its free radical scavenging and antioxidant effects, which maintains the tissue defense system against myocardial damage.     

Mobilization effects of G‐CSF,GM‐CSF,and darbepoetin‐α for allogeneic peripheral blood stem cell transplantation

The effects of GM‐/G‐CSF and darbepoetin‐α on stem cell mobilization were investigated. From February 2005 to March 2007, 30 allogeneic sibling donors were randomly assigned to a G‐CSF group (5 μg/kg/day for 5–7 days) or triple group (GM‐CSF 10 μg/kg/day on 1st and 2nd day, G‐CSF 5 μg/kg/day for 5–7 days, and darbepoetin‐α 40 mg on 1st day). The MNCs and CD34⁺ cells were not different between the two groups, although the doses (×10⁸/kg of recipient body weight) of CD3⁺ cells (3.64 ± 1.75 vs. 2.63 ± 1.36, P = 0.089) and CD8⁺ cells (1.07 ± 0.53 vs. 0.60 ± 0.30, P = 0.006) were lower in the triple group. The engraftments, frequency of RBC transfusions, and hemoglobin recovery were not different between the two groups. The cumulative incidence of overall and Grades II–IV aGVHD was 64.3% vs. 61.1% and 25.9% vs. 27.1% in the G‐CSF and triple regimen group, respectively, whereas the cumulative incidence of cGVHD was 20.8 ± 1.3% and 24.4 ± 1.7%, respectively. In conclusion, the triple regimen did not seem to be superior to G‐CSF alone in terms of the CD34+ cell dose, hemoglobin recovery, and GVHD. However, the CD8+ cell count was significantly lower in the triple regimen group. The role of a lower CD8+ cell count in the graft may need to be elucidated in the future. J. Clin. Apheresis, 2009. © 2009 Wiley‐Liss, Inc.     

Secretion and antifibrinolytic function of thrombin‐activatable fibrinolysis inhibitor from human platelets

Summary. Background: The thrombin‐activatable fibrinolysis inhibitor (TAFI) is a zymogen first characterized in human plasma that is activated through proteolytic cleavage by thrombin, thrombin in complex with thrombomodulin, or plasmin. Active TAFI attenuates fibrinolysis by removing C‐terminal lysine residues from partially degraded fibrin, thereby inhibiting a potent positive feedback loop in the fibrinolytic cascade. The existence of a separate pool of TAFI within platelets has been described. Objectives and Methods: We aimed to confirm the presence of TAFI in the medium of washed, thrombin‐stimulated platelets and to evaluate the characteristics of platelet TAFI by western blot analysis and with a quantitative assay for activated TAFI. We also assessed the ability of platelet TAFI to inhibit fibrinolysis in vitro, using a platelet‐rich thrombus lysis assay. Results: Our data are consistent with the presence of TAFI in the α‐granules of resting platelets. In contrast to previous reports, platelet TAFI is very similar in electrophoretic mobility to plasma‐derived TAFI. We also show, for the first time, that platelet‐derived TAFI is capable of attenuating platelet‐rich thrombus lysis in vitro independently of plasma TAFI. Moreover, we demonstrate additive effects on thrombolysis of platelet‐derived TAFI and TAFI present in plasma. Conclusions: Taken together, these observations indicate that the secretion of platelet‐derived TAFI can augment the concentrations of TAFI already present in plasma to enhance attenuation of the fibrinolytic cascade. This could be significant in regions of vascular damage or pathologic thrombosis, where activated platelets are known to accumulate.     

Anticonvulsive and neuroprotective effects of synergetic combination of phenytoin and gastrodin on the convulsion induced by penicillin in mice

Phenytoin (PHT) is a commonly prescribed first‐line antiepileptic drug. However, long‐term administration of PHT can cause memory loss and balance disturbance. Gastrodin (GD) is the major bioactive component in Tianma and has sedative, anticonvulsive, memory strengthening, and neuroprotective effects. To combine the two drugs seems attractive; however, little was known about the efficacy of combination therapy. In this study, convulsive attack was successfully induced by penicillin. Isobolographic analysis, memory and balance behavior test, histopathological examination, and Western blot analysis were used to investigate whether the combination therapy of GD and PHT can enhance anticonvulsive effect and reduce the side effects associated with PHT. The GD alone (950.60 mg/kg) and the PHT alone (45.50 mg/kg) could produce an anticonvulsive effect, while comparable effect could be produced by PHT : GD = 1 : 50 (8.59 : 429.27 mg/kg), which reduce the dose of PHT by 81% and GD by 55%. After the chronic anticonvulsive experiments of 16 days, the balance disturbance and short‐/long‐term memory loss were observed in the PHT group, while the PHT + GD therapy can protect the normal balance and memory function. The neuron morphology of hippocampus was preserved, and the number of surviving neurons after combination therapy was more than the model group. The amount of NF‐κB (p65) expression was increased in combination group. All above suggested the potential of the combination of PHT and GD enhances the anticonvulsive effect and the neuroprotective effect and reduces the PHT‐associated memory and balance disturbance. The PHT + GD strategy would provide new possibilities as a novel promising methodology to treat epileptic patients.