Trigonella foenum-graecum seed extract modulates expression of lipid metabolism-related genes in HepG2 cells

Objective: To investigate anti-dyslipidemic effects of hydroalcoholic fenugreek seed extracts, diosgenin, and 4-OH-Ile on HepG2 cell line. Methods: HepG2 cells were treated with hydroalcoholic fenugreek seed extracts, diosgenin, 4-OH-Ile, and orlistat. IC

Effect of crocin carotenoid on BDNF and CREB gene expression in brain ventral tegmental area of morphine treated rats

Objective: To investigate the effect of crocin carotenoid on BNDF and CREB gene expression in the brain ventral tegmental area (VTA) and the serum level of BDNF in morphine-treated rats compared to control. Methods: In this study, 40 male Wistar rats (200-250 g) were used in 5 experimental groups: 1) non morphine treat rats (control); 2) non morphine-treated rats with 25 mg/kg crocin carotenoid (i.p., for 21 d); 3) morphine treated rats (10 mg/kg twice a day, s.c., 21 d); 4 and 5) morphine-treated rats with 12.5 and 25 mg/kg crocin carotenoid, respectively. By the end of research, BDNF and CREB expression was determined by real-time-PCR method. ELISA analysis was also applied for assessing the serum BDNF level. Results: The data indicated that morphine treatment could cause a significant decrease in BDNF and CREB gene expression (P<0.01 and P<0.001, respectively) in brain VTA as well as serum level of BDNF (P<0.01) in comparison to control group. Treatment with 25 mg/kg crocin carotenoid caused a significant enhancement in BDNF and CREF gene expression (P<0.01 and P<0.05, respectively) and serum level of BDNF (P<0.01) in morphine-treated rats in comparison to morphine-treated group. Conclusions: Regarding to obtained results, crocin carotenoid can inhibit unfavorable effects of morphine on the neural system to some extent through enhancing BDNF and CREB gene expression in brain VTA and serum level of BDNF.

Anti-cancer effects of hydro-alcoholic extract of pericarp of pistachio fruits

Objective: To investigate the cytotoxicity and anti-cancer effects of hydro-alcoholic extract of pistachio pericarp on hepatocellular carcinoma cells (HepG2) and mouse fibroblast L929 cells as normal and control group cell. Methods: MTT assay was performed to investigate the cytotoxicity effects of the extract at 0-4 000 μg/mL on the cells after 24 and 48 h. The expressions of some genes involved in apoptosis including Bax, Bcl-2 and P53 were investigated by real time PCR. Results: Our results showed that after 24 and 48 hours of treatment of cells with this extract, the viability of HepG2 and L929 cells was reduced. Therefore, this extract had the cytotoxicity effect on both cells. The IC

Conditioned media derived from human adipose tissue mesenchymal stromal cells improves primary hepatocyte maintenance

Objective: Recent advances in cell therapy have encouraged researchers to provide an alternative for treatment and restoration of damaged liver through using hepatocytes. However, these cells quickly lose their functional capabilities in vitro. Here, we aim to use the secretome of mesenchymal stromal cells [MSCs] to improve in vitro maintenance conditions for hepatocytes

Materials and Methods: In this experimental study, following serum deprivation, human adipose tissue-derived MSCs [hAT-MSCs] were cultured for 24 hours under normoxic [N] and hypoxic [H] conditions. Their conditioned media [CM] were subsequently collected and labeled as N-CM [normoxia] and H-CM [hypoxia]. Murine hepatocytes were isolated by perfusion of mouse liver with collagenase, and were cultured in hepatocyte basal [William's] medium supplemented with 4% N-CM or H-CM. Untreated William's and hepatocyte-specific media [HepZYM] were used as controls. Finally, we evaluated the survival and proliferation rates, as well as functionality and hepatocyte-specific gene expressions of the cells

Results: We observed a significant increase in viability of hepatocytes in the presence of N-CM and H-CM compared to HepZYM on day 5, as indicated by MTS [3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]- 2H-tetrazolium] assay. Indocyanine green [ICG] uptake of hepatocytes in the H-CM and HepZYM groups on days 3 and 5 also suggested that H-CM maintained the hepatocytes at about the same level as the hepatocyte-specific medium. The HepZYM group had significantly higher levels of albumin [Alb] and urea secretion compared to the other groups [P<0.0001]. However, there were no significant differences in cytochrome activity and cytochrome gene expression profiles among these groups. Finally, we found a slightly, but not significantly higher concentration of vascular endothelial growth factor [VEGF] in the H-CM group compared to the N-CM group [P=0.063]

Conclusion: The enrichment of William's basal medium with 4% hAT-MSC-H-CM improved some physiologic parameters in a primary hepatocyte culture

Comparing the effects of human and fetal bovine serum on mesenchymal stem cells under oxidative stress

Objectives: Stem cells are undifferentiated cells capable of creating different types of cell in the body. Stem cell proliferation often is performed in the culture medium supplemented with Fetal Bovine Serum [FBS]. Unknown compounds in the FBS, risk of contamination and disease transmission encourages the researches toward finding an alternative to FBS. Several factors are involved in the Mesenchymal Stem Cells [MSCs] precocious death in the transplanted tissue environment. Oxidative Stress [OS] is one of the main causes of stem cell apoptosis in the initial days after transplantation. The aim of this study was to evaluate the effect of Human Serum [HS] on the viability and oxidative related enzymes in human Adipose tissue-Derived Stem Cells [ADSCs] under oxidative stress in comparison with FBS

Materials and Methods: Human serum were obtained from blood of a healthy donor persons, in respective intervals during few days. The ADSCs were isolated from lipolysis operation samples and their cuture media were supplemented with FBS or HS and different concentrations of H2O2 as the oxidative agent

Results: The results showed that cell proliferation and viability of ADSCs under oxidative stress condition was significantly higher in the culture medium supplemented with HS in comparison with FBS supplemented medium [P<0.05]

Conclusion: This study showed that FBS could be replaced by HS in MSC culture medium with improved effects on cell proliferation and oxidative related enzyme activity under oxidative Stress condition

Chemokines: small molecules participate in diabetes

Chemokines are small protein molecules involved in cell signaling processes. They play a crucial role in many physiological and pathological processes. Chemokines are functionally classified into two categories; inflammatory/inducible and constitutive. Their biologic functional differences are the result of their receptors structural differences. Recently some studies were performed about the chemokines changes in diabetes. Inflammatory mechanisms have an important role in diabetes. Inthisreviewarticle we searched the keywordschemokines, diabetes, diabetes pathogenesis, and type1 and 2diabetes in Persianresources, PubMed andfamousEnglish-language websitesthrough advanced searchenginesand found the newest studies about the roleof chemokinesin thepathogenesisof diabetes. The results of the studies showed that diabetes and its disorders enhance the activation of immune cells and the expression of cytokines such as IL-1, IL-6, IL-8, IL-10, SDF-1, INF-gamma, TGF-beta, MCP-1, IP-10, TNF-alpha, and RANTES; most of them have impact on the pathogenesis of diabetes. Comparison and analysis of the results obtained from our research and the results of performed studies in the world and Iran shows that chemokines, like other protein molecules involved in the pathogenesis and etiology of diabetes, play a role in this process

Effect of hydroalcoholic extract of capparis spinosa fruit on blood sugar and lipid profile of diabetic and normal rats

Diabetes mellitus is a common disorder of endocrine glands worldwide. Caper as a medicinal plant has anti-oxidant properties and has been used traditionally to cure diabetes. The aim of present research was to evaluate the effect of 200 and 800 mg/kg of caper fruit extract on blood sugar, glycated hemoglobin and lipid profile in diabetic and normal male rats. In this experimental study 60 rats were divided into 6 groups randomly, in which three diabetic groups received distilled water [control] and 200 and 800 mg/kg caper fruit extract respectively and three normal groups were treated as diabetic groups. The animals were received orally either water or fruit extract by gavages for 28 days. To measure the level of blood sugar, glycosylated hemoglobin and lipid profile, blood samples of animals were collected at the beginning and the end of experiment and p<0.05 was considered as significant. The blood sugar decreased in all groups receiving fruit extract compare to control groups and the decrease in blood sugar was dose dependent. Blood triglycerides decreased in all diabetic groups receiving extract compare to control but in normal rats the changes were not significant. The changes in glycated hemoglobin was not significant. Other blood metabolites that were measured in the present study were not changed significantly. The results of present study showed that consumption of Caper fruit extract lead to a significant decrease in blood sugar and also a considerable decrease in blood triglycerides in diabetic rats, therefore it seems that Caper fruit consumption may has beneficial effects on blood sugar and lipid profile

Chronic Opium Treatment Can Differentially Induce Brain and Liver Cells Apoptosis in Diabetic and Non-diabetic Male and Female Rats

It has been shown that some opium derivatives promote cell death via apoptosis. This study was designed to examine the influence of opium addiction on brain and liver cells apoptosis in male and female diabetic and non-diabetic Wistar rats. This experimental study was performed on normal, opium-addicted, diabetic and diabetic opium-addicted male and female rats. Apoptosis was evaluated by TUNEL and DNA fragmentation assays. Results of this study showed that apoptosis in opium-addicted and diabetic opium-addicted brain and liver cells were significantly higher than the both normal and diabetic rats. In addition, we found that apoptosis in brain cells of opium-addicted and diabetic opium-addicted male rats were significantly higher than opium-addicted and diabetic opium-addicted female, whereas apoptosis in liver cells of opium-addicted and diabetic opium-addicted female rats were significantly higher than opium-addicted and diabetic opium-addicted male. Overall, these results indicate that opium probably plays an important role in brain and liver cells apoptosis, therefore, leading neurotoxicity and hepatotoxicity. These findings also in away possibly means that male brain cells are more susceptible than female and interestingly liver of females are more sensitive than males in induction of apoptosis by opium.

Expression of regulated oncogen-alpha by primary hepatocytes following isolation and heat shock stimulation

High levels of regulated oncogen-alpha [GRO-a] expression have been observed in the liver. GRO-a stimulates proliferation of epithelial cells and induction of rolling and extravascular migration of neutrophils and mononuclear cells. Given the above observations, this chemokine was chosen to be analyzed in freshly isolated and cultured hepatocytes. In this study, hepatocytes [2_106 cell/ml] were isolated from male Sprague Dawley rat liver and cultured on plates that were pre-coated with collagen type-I matrix. The western and northern blot analyses were employed to detect GRO-a at the protein and mRNA levels in freshly isolated and cultured hepatocytes in response to isolation and heat shock stresses. GRO-a was shown to be expressed by isolated rat hepatocytes immediately after isolation and early culture and decreased with time. mRNA was also expressed in freshly isolated cells [0 h] and did not decrease after 48h of culture and further time points [P<0.01]. These results also demonstrated that expression of GRO-a by hepatocytes increased in response to heat shock at different time points in comparison with the control [P<0.01]. These results demonstrated that the isolation and heat shock stresses induced the expression of GRO-a in hepatocytes in a time-dependent manner. Thus, it seems that hepatocytes mimic the experiences that the liver encounters after injury in vivo. In such a situation, liver produces stress related agents like chemokines to overcome injurious conditions

Opium can differently alter blood glucose, sodium and potassium in male and female rats

To determine the effects of opium on serum glucose, potassium and sodium in male and female Wistar rat, opium solution [60 mg/kg] injected intraperitoneally and the same volume of distilled water was used as control [7 rats in each group]. Blood samples were collected at 0, 30, 60, 120, 240 and 360 minutes after injection from orbit cavity and the values of serum glucose, sodium [Na+] and potassium [K+] were measured. The data were then analyzed by the repeated measure ANOVA based on sex and case-control group. P < 0.05 considered as significant difference. Serum glucose increased significantly at 30, 60, 120 and 240 minutes after opium solution injection, in female rats compared to a control group. However, the male rats had this rise at 30, 60 and 120 minutes after opium solution injection compared to control group. While serum glucose in male rats was significantly higher than females at 30, 60 and 120 minutes, this value was higher in the female rats at 360 minutes. Therefore, serum glucose alterations following opium injection was significantly different in groups and in the sexes at different times. Sodium [Na+] rose at 60, 240 and 360 minutes significantly in all rats compared to control group. However, sodium alteration following opium injection was significantly different only between treated and control groups but sex-independent at all times. Potassium [K+] increased significantly at 60, 120, 240 and 360 minutes in male rats, compared to a control group. In female rats K+ significantly raised at 30, 120, 240 and 360 minutes. Therefore, the alteration of K+ in male and female rats was found time dependent and sex independent. According to our results, opium increased serum glucose in male and female rats differently, and it interferes with metabolic pathways differently on a gender dependent basis. Opium raised serum Na+ and K+, thus it interfere with water regulation and blood pressure via different mechanism

W-7 [a calmodulin antagonist] inhibits carrageenan-induced paw edema in intact and adrenalectomized rats

The aim of present study was to investigate the effect of W-7, a calmodulin antagonist, on a carrageenan-induced rat's paw edema. W-7[50 micro Mol/kg] was given intraperitoneally synchronous with the intraplantar injection of 0.1 ml of 0.5% carrageenan solution. After for hours, paw edema was assessed by calculating the paw volume changes which measured by hydroplethysmometer. In adrenalectomized rats, both adrenal glands were removed. Treatment of animals with W-7 reduced carrageenan-induced paw edema by 50%. In adrenalectomized rats, W-7 was also effective in reduction of paw edema [52%]. There was no significant difference between the effect of W-7 in adrenalectomized and control animals. Our findings suggest that W-7 as calmodulin inhibitor reduce carrageenan-induced paw edema and the inhibitory effect W-7 on edema formation appears not to be dependent on adrenal function

Survey of changes in complete blood count and red cell indices of whole blood incubated in vitro at different temperatures up to 48 hours

Complete blood count [CBC] is one of the most common and conventional blood test that physicians usually request. However the results of this test are affected by different factors such as, the temperature and duration of incubation, therefore the aim of this survey was to evaluate the effect of temperature and time of incubation on CBC, red blood cells [RBC] indices and white blood cells [WBC] differential count. In a cross-sectional study, blood samples were taken from 30 healthy medical students of Rafsanjan University [15 males and 15 females]. The samples divided into three parts; CBC were done on the samples up to 48 hours incubation at temperature of 25, 30, and 37°C at the time of sampling, and after 2, 8, 24 and 48 hours. Data were statistically analyzed and the following results were obtained. RBC count, hematocrit, MCH, percent of monocytes and eosinophils were constant in different temperatures, WBC count, MCHC, hemoglobin, platelets count, the percent of lymphocytes and neutrophils were constant up to 24 hours and then tend to increase with increasing temperature except lymphocytes percent that tend to decrease. MCV decreased with increasing temperature up to 8 hours and then significantly increased [from 83.89 to 87.50 fmol/1, p<0.00l]. WBC, hematocrit, MCV, platelets count, and neutrophils' percent tend to increase by the time of incubation, but RBC count, MCHC, lymphocytes' percent decreased. Hemoglobin, MCH, and the percent of monocytes and eosinophils were constant. The finding of this survey showed that some of CBC parameters can be changed with the incubation, therefore it is better to do the CBC test after blood taking as soon as possible

Extraction and purification of anti-proteinase 3 [PR3] antibodies from egg yolk

Recently it has been reported, that immunoglobulin Y [IgY] can be used instead of polyclonal antibodies extracted from mammals [IgG] for the purpose of diagnosis and therapy. These antibodies are found to have better properties in terms of specificity and ease of large-scale production. In addition, IgY binds neither to mammalian complement or Fc-receptors nor does it interfere with rheumatoid factors [RF], which has proven to be advantageous in many immunological tests. Proteinase 3 [PR3], a constituent of azurophil granules of neutrophils, is the target antigen for most anti-neutrophil cytoplasmic antibodies [c-ANCA] in Wegener granulomatosis [WG]. Capture ELISA was found to be the method of choice in case of c-ANCA determination for the diagnosis and management of WG. However, in this method, the reaction of RF with the Fc portion of IgG in capture ELISA leads to false positive in the assay of c-ANCA and is found to be the most important short-comings of available diagnostic immunochemical tests using mammalian antibodies. To avoid such unwanted interactions, laying hens were inoculated with PR3, and IgY was purified from egg yolk by acidic extraction with chloroform. The aqueous phase was treated with sodium sulphate and the precipitate collected, was dissolved in buffer and was purified using a T-gel chromatography method. The prepared IgY-anti-PR3 was used to set up a capture ELISA. Our results showed that the prepared IgY-anti-PR3 had good titer [lu g/ml in a coating system] and specificity. Hence, IgY based immunoassay would be a useful alternative to mammalian IgG antibody used in PR3 immunoassays

Transfection of 6-23 a rat thyroid carcinoma cell line, with the neuropeptide Y cDNA

Neuropeptide Y [NPY] is a 36 amino acid peptide found throughout the central and peripheral nervous system of rat and human. NPY has been proposed to play an important role in satiety. The aim of this study was to produce cell lines that secrete high levels of bioactive NPY. For this purpose, the complementary DNA [cDNA] that encodes NPY was isolated by PCR. The cDNA was then cloned into pCEP4, to form pCEP4NPY. 6-23 cells were transfected with pCEP4NPY by electroporation. Transfected cells were selected by the addition of hygromycin B to the culture medium. Resistant colonies were picked and transferred to 96-well plates. The medium was tested for IR-NPY using a specific NPY radioimmunoassay [RIA]. The IR-NPY secreted by the cells was characterized by sephadex G50 chromatography and reversed phase fast protein liquid chromatography [FPLC]. It was found to co-elute with the synthetic standard in both cases. RNA was extracted from the cells and subjected to Northern blot analysis using labeled NPY cDNA as a probe. The cells were found to express high levels of NPY at mRNA levels

Inhibition of proteinase 3 [PR3] by suramin and fetal calf serum [FCS]: effect of PR3 and suramin on Chinese hamster ovary cells [CHO-Cells]

Proteinase 3 [PR3] is a lysosomal protease that is stored in azurophilic granules neutrophilic granulocytes and monocytes. A number of inhibitors for this proteinase are reported. Comprehensive studies on the inhibitory effect of suramin and heat treated fetal calf serum [pound GFCS] on PR3 have not been reported. It has been reported that PR3 is able to destroy the cytoskeletal integral proteins, but we have not find any reports which showed the effect of this protease on Chinese hamster ovary cells [CHO-cells] in culture medium. Suramin has proven to be useful as an antitumor drug, but there was not any report on the effect of suramin on CHO-cells. The effects of various concentrations of pound GFCS [from 0.5% up to 10%] and suramin [from 0.8 pound gM up to 100 pound gM] on PR3 and different concentrations of suramin [from 0.8 pound gM up to 1000 pound gM] on CHO-cells were investigated. Data analysis were performed by, Kolmogorov-Smirnov test, ANOVA test and Tukey HSD post tests. Results showed that pound GFCS and suramin have an inhibitory effect on PR3 and these effects increased with increasing the concentration significantly [p < 0.01]. PR3 with the concentration of 2.2 Unit/ml has no effect on CHO-cells. Although suramin with the concentration of less than 125 poundgM cell growth retarded for only a few hours, but with the concentration of 125 to 250 pound gM inhibit the cell growth for a week, and after that cells gain normal growth gradually. Suramin with concentration of more than 500 pound gM inhibited the cell growth completely. Although suramin reversibly inhibit the PR3 activity but in concentration of less than 250 pound gM it had no long-term effect on CHO-cells. Therefore it can be used in the investigation of proteases. There were unknown components in pound GFCS, which cause the inhibition of PR3 activity. This finding is very important in PR3 production in culture medium. However CHO-cells are resistant to PR3 and suramin in low concentration

demonstration of polyethylenimine mediated gene transfer into the rodent hypothalamus results in persistent over-expression and phenotypic change

Polyethylenimine [PEI] has been proposed as a non-viral vector, and has been successfully used to transfer reporter genes into the central nervous system [CNS], kidneys, and lungs of adult mice. Neuropeptide Y [NPY] is a peptide expressed in the hypothalamus and is important in the regulation of body weight. Using PEI combined with stereotactic microinjection, we have successfully transferred cDNA-encoding NPY driven by the cytomegalovirus [CMV] promoter into the arcuate nucleus of adult male Wistar rats. Animals treated with NPY expressing plasmids [pNPY] gained more weight than the controls [p<0.05], with associated increases in food intake [p<0.05] and decreased brown adipose tissue activity, measured by Guanosine Diphosphate [GDP] binding to mitochondria, [p<0.05]. In a separate study, hypothalamic slices from the rats treated with pNPY/PEI showed increased NPY release [pNPY 9.7 +/- 0.3 fmol/l vs. control 8.3 +/- 0.5 fmol/l, p<0.05, n = 3]. These results suggest that PEI is an effective vector for gene transfer into the rodent brain and can increase the protein production sufficient to result a persistent phenotypic change. This technique offers the potential of a simple and effective method to manipulate gene expression localised to specific regions of the adult rodent brain