Effects of Lithium Ions on tPA-Induced Hemorrhagic Transformation under Stroke.

Thrombolytic therapy with the tissue plasminogen activator (tPA) is a therapeutic option for acute ischemic stroke. However, this approach is subject to several limitations, particularly the increased risk of hemorrhagic transformation (HT). Lithium salts show neuroprotective effects in stroke, but their effects on HT mechanisms are still unknown. In our study, we use the models of photothrombosis (PT)-induced brain ischemia and oxygen-glucose deprivation (OGD) to investigate the effect of Li+ on tPA-induced changes in brain and endothelial cell cultures. We found that tPA did not affect lesion volume or exacerbate neurological deficits but disrupted the blood-brain barrier. We demonstrate that poststroke treatment with Li+ improves neurological status and increases blood-brain barrier integrity after thrombolytic therapy. Under conditions of OGD, tPA treatment increased MMP-2/9 levels in endothelial cells, and preincubation with LiCl abolished this MMP activation. Moreover, we observed the effect of Li+ on glycolysis in tPA-treated endothelial cells, which we hypothesized to have an effect on MMP expression.

Anti-inflammatory and protective effects of Aripiprazole on TNBS-Induced colitis and associated depression in rats: Role of kynurenine pathway.

BACKGROUND: The prevalence of depression is higher in patients with inflammatory bowel disease (IBD) than in the general population. Inflammatory cytokines and the kynurenine pathway (KP) play important roles in IBD and associated depression. Aripiprazole (ARP), an atypical antipsychotic, shows various anti-inflammatory properties and may be useful in treating major depressive disorder. This study aimed to evaluate the protective effects of ARP on TNBS-induced colitis and subsequent depression in rats, highlighting the role of the KP. MATERIAL AND METHODS: Fifty-six male Wistar rats were used, and all groups except for the normal and sham groups received a single dose of intra-rectal TNBS. Three different doses of ARP and dexamethasone were injected intraperitoneally for two weeks in treatment groups. On the 15th day, behavioral tests were performed to evaluate depressive-like behaviors. Colon ulcer index and histological changes were assessed. The tissue levels of inflammatory cytokines, KP markers, lipopolysaccharide (LPS), nuclear factor-kappa-B (NF-κB), and zonula occludens (ZO-1) were evaluated in the colon and hippocampus. RESULTS: TNBS effectively induced intestinal damages and subsequent depressive-like symptoms in rats. TNBS treatment significantly elevated the intestinal content of inflammatory cytokines and NF-κB expression, dysregulated the KP markers balance in both colon and hippocampus tissues, and increased the serum levels of LPS. However, treatment with ARP for 14 days successfully reversed these alterations, particularly at higher doses. CONCLUSION: ARP could alleviate IBD-induced colon damage and associated depressive-like behaviors mainly via suppressing inflammatory cytokines activity, serum LPS concentration, and affecting the NF-κB/kynurenine pathway.

The Role of Matrix Metalloproteinases in Hemorrhagic Transformation in the Treatment of Stroke with Tissue Plasminogen Activator.

Ischemic stroke is a leading cause of disability and mortality worldwide. The only approved treatment for ischemic stroke is thrombolytic therapy with tissue plasminogen activator (tPA), though this approach often leads to a severe complication: hemorrhagic transformation (HT). The pathophysiology of HT in response to tPA is complex and not fully understood. However, numerous scientific findings suggest that the enzymatic activity and expression of matrix metalloproteinases (MMPs) in brain tissue play a crucial role. In this review article, we summarize the current knowledge of the functioning of various MMPs at different stages of ischemic stroke development and their association with HT. We also discuss the mechanisms that underlie the effect of tPA on MMPs as the main cause of the adverse effects of thrombolytic therapy. Finally, we describe recent research that aimed to develop new strategies to modulate MMP activity to improve the efficacy of thrombolytic therapy. The ultimate goal is to provide more targeted and personalized treatment options for patients with ischemic stroke to minimize complications and improve clinical outcomes.

Association of Adiponectin 45T/G (rs2241766) and Visfatin 4689G/T (rs2110385) Gene Polymorphisms with Susceptibility to Obesity.

Background: This study aimed to see whether the adiponectin 45T/G (rs2241766) and visfatin 4689G/T (rs2110385) gene polymorphisms in an Iranian population are linked to obesity and/or obesity-related traits in normal and obese individuals. Methods: 230 obese individuals and 169 healthy controls had their genomic DNA taken. The alleles and genotypes of the rs2241766 and rs2110385 polymorphisms were determined using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Results: Obese individuals had considerably greater frequencies of the G allele and GG genotypes of the rs2241766 polymorphism than healthy controls (35% vs 21%, Probability (P) <0.0001, odds ratios (OR): 1.99, 95% confidence intervals (CI): 1.45-2.75 and 21% vs 7%, P = 0.002, OR: 3.52, 95% CI: 1.81-6.85, respectively). In comparison to healthy controls, obesity patients had substantially lower frequencies of the T allele and TT genotype of the rs2241766 polymorphism (65% vs 79%, P < 0.0001, OR: 0.50, 95% CI: 0.36-0.69 and 51% vs 65%, P = 0.008, OR: 0.58, 95% CI: 0.39-0.87, respectively). Obese individuals had substantially higher frequencies of the G allele and GG genotype in the rs2110385 polymorphism than healthy controls (77% vs 69%, P = 0.01, OR: 1.47, 95% CI: 1.07-2.0 and 61% versus 51%, P = 0.047, OR: 1.5, 95% CI: 1.0-2.2, respectively). When compared to healthy controls, the frequency of the T allele in the rs2110385 polymorphism was considerably lower in obese individuals (23% vs 31%, P = 0.01, OR: 0.68, 95% CI: 0.5-0.93). Furthermore, these single nucleotide polymorphisms (SNPs) were shown to have a strong link to clinical data in obese individuals. In the case of adiponectin, 45T/G (rs2241766) genotypes, serum low-density lipoprotein, waist circumference, and diastolic blood pressure were substantially different among the rs2241766 genotypes (P = 0.007, P = 0.000, and P = 0.011, respectively). In the instance of the visfatin 4689G/T (rs2110385) gene polymorphism, serum triglycerides was substantially different among the rs2110385 genotypes (P = 0.039). Conclusions: In the Iranian population, our findings revealed a strong link between adiponectin and visfatin gene polymorphisms and obesity and several obesity-related clinical characteristics. These SNPs might be used to identify those who are at risk of becoming obese.

Oral administration of phenylalanine molecularly imprinted polymer (MIP) benefits PKU mouse model.

Phenylketonuria (PKU) is a rare genetic disorder caused by a defect in the metabolism of phenylalanine (Phe). Currently, the most commonly used treatment for PKU is dietary Phe restriction. Problems associated with Phe restricted diets include lack of universal availability, high treatment costs, and reduced adherence to continued treatment with age and finally the development of psychological and neurological problems in a significant proportion of patients despite early start of treatment. One possible approach to decreasing blood Phe level, is inhibition of GI tract absorption of this amino acid. We had previously shown that a Phe selective molecularly imprinted polymer was able to bind Phe in the GI tract and attenuate its plasma concentration. In this work, we used different orally administered Phe selective molecularly imprinted polymer doses in a PKU mouse model to further study the effects of this treatment on biochemical profile and cognitive function in test animals. Treatments started 21 days postnatally. After 3 weeks, brain and plasma amino acid profiles and brain monoaminergic neurotransmitter concentrations were measured. Behavioral profile was also evaluated. Treatment with 2% and 5% Phe selective molecularly imprinted polymer significantly reduced levels of blood Phe in PKU model animals (46% and 48% respectively) meanwhile levels of other amino acids remained unchanged. Brain dopamine concentrations in hippocampus was effectively restored by supplementation of Phe selective molecularly imprinted polymer. Finally, polymer treatment improved locomotor dysfunction in PKU model animals. Our data suggest that the Phe selective molecularly imprinted polymer can be a new candidate for treatment of PKU patients. Take home message: Orally administered Phenylalanine Selective Molecularly Imprinted Polymer is able to inhibit absorption of phenylalanine from the GI tract and may offer a new treatment, in conjunction with dietary restriction, for PKU patients.

Evaluation of Glutathione Bioactivity on Toxoplasma Gondii in BALB/c Mice Post Impact of Selenium and Calcium Supplementation.

BACKGROUND: Studies have shown that selenium is an essential component of glutathione as an important antioxidant to reduce oxidative stress and inhibit intracellular parasites' growth. In contrast, calcium in the cytosol of such parasites plays a key role in the entry of the parasite into the host cell and its primary motility. AIMS AND OBJECTIVES: The present study was designed to evaluate and compare glutathione peroxidase bioactivity effects post administration of selenium and calcium in BALB/c mice infected by Toxoplasma gondii. METHODS: Sixty BALB/c mice susceptible to T. gondii were randomly divided into twelve groups of case and control groups. There were six control groups including two positive controls infected only with the parasites either 104 or 5×104, non-infected and untreated groups. Treated controls received only calcium, selenium, or both respectively. Case groups were infected with 104 or 5×104 parasites. While each set of three case groups separately received minerals alone or together. Mice were orally fed with 200 µg selenium, 50 µg calcium or their combination for 7 days. Mice were infected by parasite's tachyzoites. Sera of mice were kept and the peritoneal macrophages were isolated for counting tachyzoites during infection. RESULTS: The results showed that selenium unlike calcium was significantly effective in reducing Toxoplasma tachyzoites compared to control groups. Moreover, glutathione peroxidase [GPX] activity was elevated in mice treated with selenium and vice versa decreased in mice treated with calcium. CONCLUSION: Administration of selenium unlike calcium reduced Toxoplasma tachyzoites proliferation by elevating bioactivity of selenium-dependent detoxification enzyme, GPX.

Association of ACE Gene Insertion/Deletion Polymorphism with Suicidal Attempt in an Iranian Population.

Deregulation of the renin-angiotensin system (RAS) plays an important role in suicide. Angiotensin converting enzyme (ACE) gene is a key component in this system. The relationship between insertion/deletion (I/D) polymorphism of ACE gene with suicide attempt (SA) is controversial. According to previous studies, allele D in this polymorphism has been considered as a potential risk factor for suicide. However, no study has been conducted in Iran to investigate this matter. This case-control study has focused on investigating the association of ACE I/D polymorphism (rs1799752) with SA in an Iranian population. The frequency of genotypes was 14% for II, 55% for ID, and 31% for DD in the case group (100 persons), and 18% for II, 74% for ID, and 8% for DD in control group (100 persons). Results show there was a significant difference in the distribution of ACE I/D polymorphism genotypes in men with SA compared to controls, as well as in women with SA compared to controls. Also, there was a significant association between DD genotype and the risk of SA compared to II genotype as reference. The severity of depression was significantly different between DD and II genotypes in SA group. According to the results, we suggest that the presence of DD genotype is possibly associated with an increased risk of SA. Maybe part of that is related to severity of depression in DD genotypes carriers of ACE I/D polymorphism.

Xanthomicrol Exerts Antiangiogenic and Antitumor Effects in a Mouse Melanoma (B16F10) Allograft Model.

Xanthomicrol, a trimethoxylated hydroxyflavone, is the main active component of Dracocephalum kotschyi Boiss leaf extract. Preliminary in vitro studies identified this compound as a potential antiangiogenic and anticancer agent. This study aimed to evaluate in vivo anticancer effect of xanthomicrol and investigate its molecular mechanism of action in a mouse melanoma (B16F10) model. Effect of xanthomicrol on B16F10 melanoma cell viability was determined using the MTT assay. For in vivo experiments, C57BL/6 mice were inoculated subcutaneously with B16F10 cells. After five days, once daily administration of xanthomicrol, thalidomide, or vehicle was commenced and continued for 21 consecutive days. On the 26th day, blood samples and tumor biopsies were taken for subsequent molecular analysis. Xanthomicrol showed inhibitory effect on viability of B16F10 melanoma cells (IC50 value: 3.433 µg/ml). Initial tumor growth, tumor volume and weight, and angiogenesis were significantly decreased in xanthomicrol-treated animals compared with those in vehicle group. Protein expression of phosphorylated Akt, mRNA expressions of HIF-1α and VEGF in tumor tissues, and serum VEGF were significantly decreased in xanthomicrol-treated animals compared with vehicle-treated animals. Thus, xanthomicrol inhibited cancer cell growth both in vitro and in vivo. This effect, at least in part, was exerted by interfering with PI3K/Akt signaling pathway and inhibiting VEGF secretion by tumor cells. Further studies are required to elucidate the exact molecular mechanisms of antitumor activity of xanthomicrol.

Evaluating the Effect of Cinnarizine on Promastigotes and Amastigotes forms of Leishmania major.

As an important global disease, cutaneous leishmaniasis is associated with complications such as secondary infections and atrophic scars. The first line treatment with antimonials is expensive and reported to have serious side effects and enhance resistance development. The main objective of this study was to evaluate the effect of Cinnarizine on standard strains of Leishmania major because of paucity of information on this subject. METHODS: In this experimental study, four concentrations of the drug (5, 10, 15 and 20 µg/ml) were added to Leishmania major cultures at 24, 48 and 72 hours intervals. MTT assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Half maximal inhibitory concentration (IC50) was ascertained by counting parasites. The inhibitory effect of the drug was compared with that of Glucantime. Flow-cytometry was performed to investigate apoptosis. Each test was repeated thrice. RESULTS: The IC50 values of Cinnarizine after 72 hours were calculated to be 34.76 µg/ml and 23.73 µg/ml for promastigotes and amastigotes, respectively. The results of MTT assays showed 48 % promastigote viability after 72 hour-exposure to Cinnarizine at 20 µg/ml concentration. Programmed cell death in promastigote- and amastigote-infected macrophages was quantified to be 13.66 % and 98.7 %, respectively. Flow- cytometry analysis indicated that Cinnarizine induced early and late apoptosis in parasites. All treatments produced results which differed significantly from control group (P<0.05). CONCLUSION: Cinnarizine showed low toxicity with anti-leishmanial and apoptosis effects on both promastigote and intracellular amastigote forms. Therefore, we may suggest further assessment on animal models of this drug as candidates for cutaneous leishmaniasis therapy.

Gender-related relation between metabolic syndrome and S447X and HindIII polymorphisms of lipoprotein lipase gene in northern Iran.

BACKGROUND: Metabolic syndrome is a cluster of conditions that increase risk of cardiovascular morbidity and mortality. Among genetic factors that contributed to incidence of metabolic syndrome, Polymorphisms of Lipoprotein lipase (LPL) are major candidates especially because of their effect on obesity and dyslipidemia. S447X (rs328) and Hind III (rs320) Polymorphisms of LPL gene have been reported to change LPL activity, resulting in altered triglyceride (TG) and high density lipoprotein Cholesterol (HDL-C) levels. This study investigates the effects of these gene polymorphisms on factors affecting metabolic syndrome in northern population of Iran. METHODS: Studied population included 223 adults consisting 90 women and 133 men with body mass index (BMI) ≥ 30 kg/m2 as obese subjects, and 156 healthy participants as a control group with BMI <25 that included 68 women and 88 men. All factors causing metabolic syndrome were evaluated. Also DNA was extracted from blood samples and HindIII and S447X LPL gene polymorphisms were screened by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). CONCLUSIONS: The present study proves that some genotypes of S447X were associated with a reduced risk of developing low HDL-C only in men, while the protective effects of HindIII on hypertriglyceridemia were only seen in women [corrected]. The point is that this relation is affected by the weight profile of the participants. It can be concluded that there is a gender-related relation between the polymorphisms of LPL gene and the risk factors for incidence of metabolic syndrome in the northern population of Iran.

Effect of 2-Phenylethanol as Antifungal Agent and Common Antifungals (Amphotericin B, Fluconazole, and Itraconazole) on Candida Species Isolated from Chronic and Recurrent Cases of Candidal Vulvovaginitis.

The antifungal effects of 2-phenylethanol are clearly visible through its intervention in Candida morphogenesis. Chronic and recurrent vulvovaginitis, however, does not respond to this standard experimental therapy; therefore, the study presented in this article investigated the effect of common antifungal drugs (amphotericin B [AMB], fluconazole [FLU], and itraconazole [ITC]), in combination with 2-phenylethanol, on the Candida species isolated from cases of chronic and recurrent vulvovaginitis, thereby allowing the recommendation of a more appropriate treatment option. Forty isolates from patients with chronic and recurrent vaginal candidiasis were investigated in this experimental study. The specimens were examined by direct microscopy, culturing, and PCR to identify the species. The antifungal effects of 2-phenylethanol and conventional drugs, both alone and in combination, were determined in duplicate. Finally, the findings were analyzed. In this study, 40 strains of Candida species were identified, whose agents were Candida albicans (95%) and Candida africana (5%). After 48 h, the minimum inhibitory concentration (MIC) range of the 2-phenylethanol was 800-3,200 µg/mL. Also, in the final study on the MIC levels of common antifungal drugs, AMB (0.42 µg/mL) had the lowest MIC, FLU (40.51 µg/mL) had the highest MIC, and the combination of ITC and 2-phenylethanol had the lowest fractional inhibitory concentration index (FICI) of any of the combinations (FICI range, 0.26-1.03). Combining FLU and ITC with 2-phenylethanol can effectively increase their antifungal effect.

Commentary on prevention a possible drug-drug interaction: is concurrent administration of orlistat and pioglitazone increase the risk of durg-induced hepatotoxicity?

BACKGROUND: Drug-drug interactions (DDIs) are an emerging threat to public health and are difficult to detect. To prevent DDIs and their burden, the possible DDIs should be kept in mind. We know that the obesity predisposes to the development of insulin resistance and type 2 diabetes. Therefore, combinational uses of antiobesity drugs and glucose-lowering drugs are very common. As the hepatotoxicity of both pioglitazone (an antidiabetic drug) and orlistat (an antiobesity drug) has been shown in some cases, the aim of this study was to evaluate the interaction of pioglitazone and orlistat in human hepatocellular cell line human hepatocellular carcinoma (HepG2) cells to determine their effect on liver toxicity. METHODS: Human hepatocellular carcinoma cells were treated with 25 µM Pioglitazon (Pio), 20 µM Orlistat (Orl) pioglitazone, orlistat or combination of them. The MTT assay was used to assess cell viability. RESULTS: Pioglitazone and orlistat combination caused a loss of HepG2 cell viability. While pioglitazone (25 µM) and orliatat (20 µM) alone decreased the cell viability around 91% and 85% respectively (notsignificant, P > 0.05), the combination of these two drugs reduced the amount of viable cells to 55% which was significant when compared with each drug alone (P < 0.001). CONCLUSIONS: Revealing the significant loss of viability of HepG2 cells in the combination use of pioglitazone and orlistat indicates these two drugs should not be administered at the same time to prevent their hepatotoxic effects especially in patients with liver dysfunction.

Changes in bone biological markers after treatment of Iranian diabetic patients with pioglitazone: No relation to polymorphism of PPAR-γ (Pro12Ala).

BACKGROUND: Thiazolidinediones (TZDs) improves insulin sensitivity by activating the peroxisome proliferator-activated receptor γ (PPAR-g). We aimed to study any association between variation in bone biochemical markers and single nucleotide polymorphism (SNP) in PPAR-γ (Pro12Ala) and investigate if these genetic variants affect bone turnover markers in Iranian diabetic population before and after treatment with pioglitazone. MATERIALS AND METHODS: A total of 101 patients (type 2 diabetic (T2D) were treated for 12 weeks with pioglitazone (15 mg/day). Bone Biological markers, osteocalcin, and C-terminal telopeptide of type 1 collagen (CTx) were measured before and after pioglitazone therapy. We genotyped 128 nondiabetic controls and 101 T2D patients as well. Pro12Ala polymorphism in PPAR-γ was done by real-time polymerase chain reaction (RT-PCR) using TaqMan assay. RESULTS: There were statistically significant differences in allele frequencies of Pro12Ala while comparing the controls with T2D subjects. Ala frequency was 7 vs 3%, P = 0.036 and genotypic frequency of Pro/Ala was 5.94 vs 14.06%, P = 0.04. After treatment, the homeostasis model of assessment of insulin resistance (HOMA-IR) as a maker of insulin resistance was significantly decreased (P < 0.001). In respect of bone turnover markers, CTx values decreased and osteocalcin significantly increased. (P < 0.001). CONCLUSION: Our findings did not reveal a significant association between this polymorphism and bone turnover markers after pioglitazone treatment. The reduced insulin resistance might be the reason that CTx values decreased and osteocalcin increased significantly after short-term pioglitazone treatment. These findings suggest the need for further studies on the possible role of insulin in regulation of bone metabolism.

In vitro selection and characterization of deoxyribonucleic acid aptamers for digoxin.

The low therapeutic index of digoxin necessitates careful monitoring of its serum levels. Most of digoxin immunoassays suffer from interferences with digoxin-like immunoreactive substances. Since aptamers have been shown to be highly specific for their targets, the aim of this study was to develop DNA aptamers for this widely used cardiac glycoside. Digoxin was coated onto the surface of streptavidin magnetic beads. DNA aptamers against digoxin were designed using Systematic Evolution of Ligands by Exponential enrichment method (SELEX) by 11 iterative rounds of incubation of digoxin-coated streptavidin magnetic beads with synthetic DNA library, DNA elution, electrophoresis and PCR amplification. The PCR product was cloned and sequenced. Binding affinity was determined using digoxin-BSA conjugate, coated onto ELISA plate. Inhibitory effect of anti-digoxin aptamer was conducted using isolated guinea-pig atrium. Three aptamers (D1, D2 and D3) were identified. Binding studies of fluorescein-labeled truncated (without primer binding region) D1 and D2 and full length D1 anti-digoxin aptamers were performed and their corresponding dissociation constants values were 8.2×10(-9), 44.0×10(-9) and 17.8×10(-9) M, respectively. This is comparable to what other workers have obtained for interaction of monoclonal antibodies raised against digoxin. There was little difference in binding affinity between full length and truncated anti-digoxin D1 aptamer. D1 anti-digoxin aptamer also inhibited the effects of digoxin on the isolated guinea-pig atrium. D1 anti-digoxin aptamer distinguished between digoxin and ouabain in both tissue study and binding experiments. Our finding indicated that D1 anti-digoxin aptamer can selectively bind to digoxin. Further studies might show its suitability for use in digoxin assays and as a therapeutic agent in life-threatening digoxin toxicity.

Role of endogenous vitamin E in renal ischemic preconditioning process: differences between male and female rats.

BACKGROUND: Antioxidants such as α-tocopherol (vitamin E) and ß-carotene (vitamin A) play an important role in protective effect of repeated brief periods of ischemia, namely ischemic preconditioning (IPC). Values of these antioxidants were investigated and compared after induction of ischemia reperfusion (IR) and kidney IPC in both male and female rats. METHODS: Forty eight Wistar rats were divided randomly into six groups of 8: groups A and B (male and female controls, respectively), group C (male IR or IR cases), group D (female IR cases) and groups E and F (male and female IPC cases, respectively). In groups C and D, ischemia was induced by clamping of left renal arteries for 45 min. In groups E and F, rats underwent four cycles of 4 min of arterial clamping and 11 min of de-clamping before final 45 min ischemia induction. Afterward, serum was collected to assess the blood urea nitrogen, creatinine and vitamins E and A values. Renal tissues were obtained for histological assessments. RESULTS: α-tocopherol levels in male and female rats showed a significant increase in IPC compared with IR group (P<0.01) and also in female IPC compared with male IPC group. ß-carotene levels had no significant variations. Histological evaluation showed that IR-induced renal injuries were less in female rats. Also, protective effects of IPC were more in female rats (P<0.01). CONCLUSIONS: Renal IPC reduced damages in both male and female rats, but tissue injuries in females were decreased much more along with the increase of endogenous vitamin E.

The association between adiponectin (+45T/G) and adiponectin receptor-2 (+795G/A) single nucleotide polymorphisms with cirrhosis in Iranian population.

Adiponectin which possesses anti-inflammatory and insulin-sensitizing properties is elevated in blood circulation of liver cirrhosis patients. The genetic variations in the adiponectin gene can affect the circulating adiponectin level and stimulation of adiponectin receptor that may affect the activity of adiponectin. We investigated the effect of adiponectin single nucleotide polymorphisms (SNP) 45 T/G and adiponectin receptor-2 gene SNP 795G/A in cirrhotic Iranian population. A total of 97 cirrhotic patients and 128 healthy controls from Iranian population were genotyped for the adiponectin and adiponectin receptor 2 gene (+45T>G and 795G/A) by polymerase chain reaction-restriction fragment length polymorphism. G frequency was 21.1% versus 12.89% (P = 0.001) for SNP45, and G frequency was 75.8% versus 76.2% (P = 0.526) for SNP795G/A in the patients and control group, respectively. Based on our findings, the expression of the G allele at SNP45 is higher in the patient group compared with healthy subjects, suggesting that it may affect liver injury through changes in the plasma adiponectin level.

Polymorphism of adiponectin (45T/G) and adiponectin receptor-2 (795G/A) in an Iranian population: relation with insulin resistance and response to treatment with pioglitazone in patients with type 2 diabetes mellitus.

Adiponectin, an adipose-derived plasma protein, is reduced in patients with obesity and type 2 diabetes. Thiazolidinediones can increase adiponectin levels and improve insulin sensitivity. This study investigated the associations between type 2 diabetes and two single-nucleotide polymorphisms in the adiponectin (45T/G) and adiponectin receptor-2 gene (795G/A), and investigated whether these genetic variants affect the response to pioglitazone in Iranian patients with type 2 diabetes. We genotyped 128 non-diabetic participants and 101 patients with type 2 diabetes for 45T/G and 795G/A with polymerase chain reaction-restriction fragment length polymorphism assays. Patients were treated with pioglitazone for 12 weeks, after which we compared laboratory parameters in these two groups. Fasting blood sugar differed significantly in individuals with different 795G/A genotypes after pioglitazone treatment (P = 0.009). The mean decrease in insulin/glucose ratio after treatment also differed significantly in individuals with different 45T/G genotypes (P = 0.035). The T allele frequency for 45T/G was 87.11% in controls versus 81.68% in patients (P = 0.071). The TG and GG genotypes were more frequent in patients (P = 0.032). The G allele frequency for 795G/A was 76.17% in controls versus 80.20% in patients (P = 0.179). 795G/A variants were not significantly different between patient and control group. The adiponectin gene 45T/G mutation may be an important determinant of type 2 diabetes in the Iranian population. However, adiponectin 45T/G and adiponectin receptor-2 795G/A polymorphisms were not significantly associated with the response to pioglitazone in our sample.

Genotyping of peroxisome proliferator-activated receptor gamma (PPAR-γ) polymorphism (Pro12Ala) in Iranian population.

BACKGROUND: The peroxisome proliferator-activated receptor-γ (PPAR-γ) is a nuclear hormone receptor. It is predominantly expressed in adipose tissue and as a receptor for thiazolidinediones, it has drawn attentions towards itself as a key molecule to trigger pathways involving in some diseases such as cancers, type 2 diabetes, inflammations and osteoporosis. A proline changed to alanine in codon 12 of PPAR-γ gene (Pro12Ala) has been known to be responsible for decreased risk of type 2 diabetes. The aim of the present study is to investigate the frequency of Pro12Ala polymorphism in PPAR-γ in healthy Iranian population to compare with other populations. Understanding this polymorphism may help us in better diagnosis, prevention, and therapeutic approaches toward a better management of diseases such as type 2 diabetes and osteoporosis. METHODS: 128 healthy volunteers were enrolled in this study. To determine single nucleotide polymorphisms (SNPs), we did real time polymerase chain reaction (RT-PCR), using TaqMan allelic discrimination assays. RESULTS: Genotype frequencies for PPAR-γ gene Pro12Ala (rs1801282) polymorphism were 0.86 for CC, 0.14 for CG, 0.00 for GG while allelic frequencies were 0.93 and 0.0.07 for C and G, respectively. CONCLUSIONS: There are statistical differences between the distribution of the PPAR-γ-2 Pro12Ala polymorphism in other populations and Iranian population.

Polymorphism of peroxisome proliferator-activated receptor γ (PPARγ) Pro12Ala in the Iranian population: relation with insulin resistance and response to treatment with pioglitazone in type 2 diabetes.

The peroxisome proliferator-activated receptor γ (PPARγ) has important effects on insulin sensitivity, obesity and diabetes. Pioglitazone improves insulin sensitivity by activating PPARγ. In view of inter-individual variability in therapeutic response to pioglitazone, this study was designed to search for an association between type 2 diabetes mellitus and Pro12Ala single-nucleotide polymorphism (SNP) in PPARγ (SNP rs1801282) and to investigate whether these genetic variants affect pioglitazone response in an Iranian population. A total of 101 patients with type 2 diabetes were treated for 12 weeks with pioglitazone (15 mg/day). Paraclinical parameters were measured before and after therapy. We genotyped 128 control participants without diabetes and all patients with type 2 diabetes. The Pro12Ala polymorphism in PPARγ was detected with real-time PCR. The Ala allele was found in 7% of the control participants vs. 3% of those with type 2 diabetes (P=0.04). The genotypic frequencies of Pro/Ala were 14.06% in the former group vs. 5.94% in the latter (P=0.036). There were significant changes in some laboratory values and biochemical markers of insulin sensitivity after pioglitazone therapy. The Pro12Ala polymorphism was associated with significant changes in insulin-to-glucose ratio after treatment (P=0.015 and P=0.005). Our findings suggest that in carriers of the 12Ala variant, pioglitazone significantly reduced the risk of type 2 diabetes, and in diabetic patients with the Pro12Ala genotype, the therapeutic response to treatment was better than in patients with the Pro12Pro genotype, although the difference between groups did not reach statistical significance.