scholarly journals Mass spectrometric analysis of rat liver cytosolic glutathione S-transferases: modifications are limited to N-terminal processing

1995 ◽  
Vol 308 (1) ◽  
pp. 69-75 ◽  
Author(s):  
H I Yeh ◽  
C H Hsieh ◽  
L Y Wang ◽  
S P Tsai ◽  
H Y Hsu ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Female Rats ◽  
Spectrometric Analysis ◽  
Affinity Column ◽  
Female Rat ◽  
Significant Difference ◽  
Cytosolic Glutathione ◽  
Glutathione S Transferases ◽  
Molecular Masses ◽  
Rat Livers

Cytosolic glutathione S-transferases (GSTs) from rat livers were purified using an S-hexylglutathione affinity column. The GST subunits were resolved by reverse-phase HPLC and their molecular masses were determined by electrospray mass spectrometry. The major hepatic GSTs detected were subunits 1, 1′, 2, 3 and 4, with molecular mass of 25,520, 25,473, 25,188, 25,782 and 25,571 Da respectively. Subunits 6, 7 and 10 are minor components, with molecular mass of 25,551, 23,308 and 25,211 Da respectively. Alternatively, the hepatic GSTs were purified using a glutathione affinity column. Subunits 1, 1′, 2, 8 and 10 were eluted from this column with GSSG, the oxidized form of glutathione. Subunit 8 has a molecular mass of 25,553 Da. The remaining proteins on the glutathione affinity column were removed with glutathione and S-hexylglutathione. Subunits 2, 3, 4 and 6 could be detected in the eluate. We could not detect any significant difference in molecular mass between GSTs isolated from male and female rat livers. Cytosolic GSTs were isolated from livers of buthionine sulphoximine-treated female rats for MS analysis. The molecular masses obtained were identical to those determined for the controls.

scholarly journals Mass spectrometric analysis of rat ovary and testis cytosolic glutathione S-transferases (GSTs): identification of a novel class-Alpha GST, rGSTA6*, in rat testis

1997 ◽  
Vol 323 (2) ◽  
pp. 503-510 ◽  
Author(s):  
Cheng-Hsilin HSIEH ◽  
Shu-Ping TSAI ◽  
Horng-I YEH ◽  
Tsuey-Chyi SHEU ◽  
Ming F. TAM
Keyword(s):  
Peptide Sequencing ◽  
Spectrometric Analysis ◽  
Sequencing Analysis ◽  
Post Translational Modifications ◽  
Rat Testis ◽  
Cytosolic Glutathione ◽  
Glutathione S Transferases ◽  
Molecular Masses ◽  
Rat Ovary ◽  
Internal Peptide

Cytosolic glutathione S-transferases (GSTs) from rat ovaries and testis were purified by a combination of GSH and S-hexylglutathione affinity chromatography. The isolated GSTs were subjected to reverse-phase HPLC, electrospray MS and N-terminal peptide sequencing analysis. The major GST isoenzymes expressed in ovaries are subunits A3, A4, M1, M2 and P1. Other isoenzymes detected are subunits A1, M3 and M6*. In rat testis, the major GST isoenzymes expressed are subunits A3, M1, M2, M3, M5* and M6*. Subunits A1, A4 and P1 are expressed in lesser amounts. We could not detect post-translational modifications of any GSTs with known cDNA sequence. The molecular masses of subunits M5* and M6*, two class-Mu GSTs that have not been cloned, were determined to be 25495 and 26538 Da respectively. An N-terminally modified protein from rat testis with molecular mass 25737 Da was isolated from the S-hexylglutathione column. Results from internal peptide sequencing analysis indicate that this is a novel class-Alpha GST that has not been previously reported. We designate this protein rGSTA6*.

scholarly journals Sex differences in the hydroxylation of cholecalciferol and of 5 β-cholestane-3 α, 7 α, 12 α-triol in rat liver

1987 ◽  
Vol 247 (1) ◽  
pp. 73-78 ◽  
Author(s):  
K Saarem ◽  
J I Pedersen
Keyword(s):  
Sex Hormones ◽  
Rat Liver ◽  
Female Rats ◽  
Regulatory Control ◽  
Male Rats ◽  
Male Rat ◽  
Female Rat ◽  
Significant Difference ◽  
Alpha 7 ◽  
Rat Livers

The effect of sex hormones on hydroxylation of cholecalciferol (‘vitamin D3’) and of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol has been investigated in female- and male-rat livers. The mitochondrial cholecalciferol 25-hydroxylase and C27-steroid 27-hydroxylase activities were respectively 4.6- and 2.7-fold higher in female- than in male-rat livers. The microsomal 1 alpha-hydroxycholecalciferol 25-hydroxylase was 2.8-fold higher in male- than in female-rat liver. No significant difference was found in the microsomal 25-hydroxylation of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol. Liver microsomes (microsomal fractions) from male, but not from female, rats also catalysed 1-hydroxylation of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol. Injection of testosterone into female rats decreased the mitochondrial cholecalciferol 25-hydroxylase and C27-steroid 27-hydroxylase activities, but not to a statistically significant extent. Testosterone treatment had no effect on the microsomal hydroxylases in female-rat liver. Injection of oestradiol valerate to male rats resulted in increased activities of both mitochondrial hydroxylases to the same levels as those of control females, while the microsomal enzyme activities decreased. The present results indicate that sex hormones exert a regulatory control on the mitochondrial cholecalciferol 25-hydroxylase and C27-steroid 27-hydroxylase activities.

scholarly journals Effect of methanolic extract of Amaranthus viridis leaves on reproductive functions in wistar female rats

2019 ◽  
Vol 9 (6-s) ◽  
pp. 119-126
Author(s):  
Mataphouet Emmanuel AFFY ◽  
Wahon Marie-Odile TOVI ◽  
N’guessan Ernest ZOUGROU ◽  
Koffi KOUAKOU
Keyword(s):  
Methanolic Extract ◽  
Day Treatment ◽  
Total Duration ◽  
Female Rats ◽  
Histological Structure ◽  
Female Rat ◽  
Pharmacological Effects ◽  
Vaginal Smears ◽  
Estrogenic Potential ◽  
Significant Difference

Objective : The aim of this study is to determine the pharmacological effects and the estrogenic properties of Amaranthus viridis leaves on the reproductive function of animal model (female rat). Methods : Vaginal smears performed 9 days before treatment allowed to select female rats having alternated on two cycles a regularity. Thereafter, the selected rats were administered by gavage daily for 28 days taking care of smear every morning at 7am from the first day of treatment follow the evolution of the cycle. For this study 20 nulliparous rats, 2 months old, weighing between 120-150 g. The first group (control) was administered with olive oil and the other three batches received respectively the doses 200, 400 and 600 mg/kg of the methanolic extract of Amaranthus viridis. At the end of the 28-day treatment, ovary and uterine horn were removed, histological and hormonal parameters were studied for determine pharmacological effects of methanolic extract of Amaranthus viridis. Results : The extract caused a disturbances of the cycle according to the doses administered. Disturbances at doses 200 and 400 mg/kg PC are significant. The calculation of the total duration of the different phases of the cycle revealed very significant increases in the estrous phase (P<0.01) by 22.79 % and 17.13 % at the respective doses of 200 and 400 mg/kg b.w compared to control. Non-significant difference was recorded on FSH, LH and estradiol level. On progesterone level, administration of the methanolic extract showed a significant difference at dose of 600 mg/kg b.w compared to control. On histological structure of the ovary, the presence of active and degenerate corpus luteum, secondary follicles depending on the dose administered were recored. Conclusion : The results showed that the methanolic extract of Amaranthus viridis contain estrogenic substances or estrogen-like substances according to a dose-dependent mechanism, with high estrogenic potential at doses of 200 and 400 mg/kg b.w . Keywords: vaginal smears, Amaranthus viridis, methanolic extract, histology

scholarly journals Sex-Related Liver Injury Due to Alcohol Involves Activation of Kupffer Cells by Endotoxin

2000 ◽  
Vol 14 (suppl d) ◽  
pp. 129D-135D ◽  
Author(s):  
Ronald G Thurman
Keyword(s):  
Liver Injury ◽  
Kupffer Cells ◽  
Coupling Constants ◽  
Female Rats ◽  
Male Rat ◽  
Gadolinium Chloride ◽  
Female Rat ◽  
Ethanol Treatment ◽  
Necrosis Factor Alpha ◽  
Rat Livers

Females have a greater susceptibility to ethanol-induced liver injury than males. Females who drink ethanol regularly and have been overweight for 10 years or more are at greater risk for both hepatitis and cirrhosis than males, and females develop ethanol-induced liver injury more rapidly and with less ethanol than males. Female rats on an enteral ethanol protocol exhibit injury more quickly than males and have widespread fatty changes over a larger portion of the liver lobule. Moreover, levels of plasma endotoxin, intracellular adhesion molecule-1, free radical adducts, infiltrating neutrophils and nuclear factor kappa B are doubled in female rat livers compared with male rat livers after enteral ethanol treatment. Additionally, estrogen treatment in vivo increases the sensitivity of hepatic macrophages or Kupffer cells to endotoxin. Evidence has been presented that Kupffer cells are pivotal in the development of ethanol-induced liver injury. Destroying Kupffer cells with gadolinium chloride or decreasing bacterial endotoxin by sterilizing the gut with antibiotics inhibits early inflammation due to ethanol. Similar results have been obtained with anti-tumour necrosis factor-alpha antibody. These data pointed to the hypothesis that ethanol-induced liver injury involves elevations in circulating endotoxin concentrations leading to activation of Kupffer cells, which causes a hypoxia-reoxygenation injury. This theory has been tested using pimonidazole, a 2-nitroimidazole marker, to quantify hypoxia in downstream, pericentral regions of the hepatic lobule. After chronic enteral ethanol treatment, pimonidazole binding doubles. Enteral ethanol also increases free radicals detected with electron spin resonance. Radical adducts, with coupling constants such as alpha-hydroxyethyl radical, have been shown to arise from ethanol. Importantly, hypoxia and radical production detected in bile are also decreased by the destruction of Kupffer cells with gadolinium chloride. These data support the hypothesis that Kupffer cells contribute to the vital sex differences in liver injury caused by ethanol.

scholarly journals Isolation and Quantification of Sphingosine and Sphinganine from Rat Serum Revealed Gender Differences

Biomolecules ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 459 ◽  
Author(s):  
Graham Brogden ◽  
Diab M. Husein ◽  
Pablo Steinberg ◽  
Hassan Y. Naim
Keyword(s):  
High Performance ◽  
Gene Mutations ◽  
Female Rats ◽  
Sphingolipid Metabolism ◽  
Female Rat ◽  
Serum Samples ◽  
Fluorescence Detector ◽  
Rat Serum ◽  
Male And Female Rats ◽  
Significant Difference

Sphingolipids are an important group of lipids that play crucial roles in living cells, facilitating cell recognition, signal transduction and endocytosis. The concentration of sphingosine and some of its derivatives like sphinganine may serve as a biomarker for the diagnosis of sphingolipidoses or be used for further research into similar diseases. In this study, a sphingolipid extraction and a high resolution detection method specific for sphingosine and sphinganine was adapted and tested. Lipids were extracted from rats’ serum, coupled to o-phthalaldehyde and detected with a fluorescence detector after running through a silica gel column in a high performance liquid chromatography system. With this method, we analysed 20 male and 20 female rat serum samples and compared the concentrations of sphingosine and sphinganine. The results showed a significant difference between the sphingosine concentrations in the male and female rats. The sphingosine concentration in female rats was 805 ng/mL (standard deviation, SD ± 549), while that in males was significantly lower at (75 ng/mL (SD ± 40)). Furthermore, the sphingosine:sphinganine ratio was almost 15-fold higher in the females’ samples. The method presented here facilitates the accurate quantification of sphingosine and sphinganine concentrations down to 2.6 ng and 3.0 ng, respectively, and their ratio in small amounts of rat serum samples to study the sphingolipid metabolism and its potential modulation due to gene mutations or the effect of prevalent toxins.

scholarly journals Rat kidney glutathione S-transferase 1 subunits have C-terminal truncations

1996 ◽  
Vol 314 (3) ◽  
pp. 1017-1025 ◽  
Author(s):  
Horng-I. YEH ◽  
Jing-Yu LEE ◽  
Shu-Ping TSAI ◽  
Cheng-Hsilin HSIEH ◽  
Ming F. TAM
Keyword(s):  
Rat Liver ◽  
Peptide Mapping ◽  
Rat Kidney ◽  
Cdna Sequencing ◽  
Sequencing Data ◽  
Glutathione S Transferase ◽  
Reverse Phase Hplc ◽  
Cytosolic Glutathione ◽  
Glutathione S Transferases ◽  
Molecular Masses

Cytosolic glutathione S-transferases (GSTs) from rat kidneys were purified by a combination of glutathione and S-hexylglutathione affinity columns. The isolated GSTs were subjected to reverse-phase HPLC and electrospray MS analysis. The major GST isoenzymes expressed in kidney are subunits 1, 2, 7 and 8. GST 1´, 3, and 4 are expressed in minor amounts. GST 10 is barely detectable in the male kidney cytosol. The molecular masses of these rat kidney GST subunits were determined by MS. The values obtained for subunits 1´, 2, 3, 4, 7, 8 and 10 are identical with those obtained for rat liver GSTs. Rat kidney GST 1 consists of three polypeptides, with molecular masses of 25517, 25372 and 24982 Da. Results from peptide mapping, MS and amino-acid-sequencing analyses indicate that the major components were generated by deleting the C-terminal phenylalanine (24982 Da) and the C-terminal IFKF tetrapeptide (25372 Da) from the GST 1 subunit, respectively. The 1-chloro-2,4-dinitrobenzene-conjugating and peroxidase activities of kidney GST 1 are substantially lower than for its counterpart from rat liver. In addition, rat kidney GST 1 has an arginine and a valine residue at positions 151 and 207 respectively. The results are in contradiction with the SWISS-PROT and GenBank rat liver GST 1 cDNA-sequencing data, which give a lysine and a methionine at the corresponding positions. Further analyses indicate that rat liver GST 1 also has C-terminal phenylalanine deletion, and an arginine and a valine residue at positions 151 and 207 respectively. However, the C-terminal-tetrapeptide-deleted form was not observed for rat liver GST 1.

scholarly journals Mode of GH administration and gene expression in the female rat brain

2017 ◽  
Vol 233 (2) ◽  
pp. 187-196 ◽  
Author(s):  
Marion Walser ◽  
Linus Schiöler ◽  
Jan Oscarsson ◽  
Maria A I Åberg ◽  
Ruth Wickelgren ◽  
...  
Keyword(s):  
Mixed Model ◽  
Brain Regions ◽  
Female Rats ◽  
Male Rats ◽  
Male Rat ◽  
Sexually Dimorphic ◽  
Female Rat ◽  
Expression Levels ◽  
Mixed Model Analysis ◽  
Significant Difference

The endogenous secretion of growth hormone (GH) is sexually dimorphic in rats with females having a more even and males a more pulsatile secretion and low trough levels. The mode of GH administration, mimicking the sexually dimorphic secretion, has different systemic effects. In the brains of male rats, we have previously found that the mode of GH administration differently affects neuron–haemoglobin beta (Hbb) expression whereas effects on other transcripts were moderate. The different modes of GH administration could have different effects on brain transcripts in female rats. Hypophysectomised female rats were given GH either as injections twice daily or as continuous infusion and GH-responsive transcripts were assessed by quantitative reverse transcription polymerase chain reaction in the hippocampus and parietal cortex (cortex). The different modes of GH-administration markedly increased Hbb and 5′-aminolevulinate synthase 2 (Alas2) in both brain regions. As other effects were relatively moderate, a mixed model analysis (MMA) was used to investigate general effects of the treatments. In the hippocampus, MMA showed that GH-infusion suppressed glia- and neuron-related transcript expression levels, whereas GH-injections increased expression levels. In the cortex, GH-infusion instead increased neuron-related transcripts, whereas GH-injections had no significant effect. Interestingly, this contrasts to previous results obtained from male rat cortex where GH-infusion generally decreased expression levels. In conclusion, the results indicate that there is a small but significant difference in response to mode of GH administration in the hippocampus as compared to the cortex. For both modes of GH administration, there was a robust effect on Hbb and Alas2.

scholarly journals Benoxacor Is Enantioselectively Metabolized By Rat Liver Subcellular Fractions

2020 ◽  
Author(s):  
Derek Simonsen ◽  
David M. Cwiertny ◽  
Hans-Joachim Lehmler
Keyword(s):  
Environmental Fate ◽  
Female Rats ◽  
Male Rats ◽  
Male Rat ◽  
Cytochrome P450 Enzymes ◽  
Transport Studies ◽  
Glutathione S Transferases ◽  
Mammalian Toxicity ◽  
Rat Livers

This study investigated the enantioselective metabolism of benoxacor, an ingredient of herbicide formulations, in rats in vitro. Benoxacor was incubated for ≤ 30 minutes with microsomes or cytosol from female or male rat livers, and its enantioselective depletion was measured using gas chromatographic methods. Benoxacor was depleted in incubations with active microsomes, suggesting its metabolism by hepatic cytochrome P450 enzymes. In the presence of glutathione, benoxacor was also depleted in cytosolic incubations, consistent with its metabolism by glutathione S-transferases. The depletion of benoxacor was faster in incubations with cytosols from male than female rats, whereas no statistically significant sex-differences was observed in microsomal incubations. Microsomal incubations showed an enrichment of the first eluting benoxacor enantiomer (E<sub>1</sub>-benoxacor). A greater enrichment occurred in incubations with microsomes from female (EF=0.67±0.01) than male rats (EF=0.60±0.01). Cytosolic incubations from female rats resulted in enrichment of E<sub>1</sub>-benoxacor (EF=0.54±0.01), while cytosolic incubations from male rats displayed enrichment of the second eluting enantiomer (E<sub>2</sub>-benoxacor; EF=0.43±0.01). Sex-dependent differences in the metabolism of benoxacor in rats could significantly impact ecological risks and mammalian toxicity. Our results also suggest changes in the enantiomeric enrichment of benoxacor may be a powerful tool for source apportionment and environmental fate and transport studies. <br>

Effects of kolaviron on pneumonia-like infection induced in albino wistar rats.

2020 ◽  
Vol 19 ◽  
Author(s):  
Dozie-Nwakile Ogechukwu Calista ◽  
Nwakile Calistus Dozie ◽  
Uchendu Ikenna Kingsley ◽  
Okonkwo Francis Catherine ◽  
Onyemelukwe Ngozi Felicia
Keyword(s):  
Wistar Rats ◽  
Female Rats ◽  
Broth Culture ◽  
Male Rat ◽  
Klebsiella Pneumonia ◽  
Female Rat ◽  
Once Daily ◽  
Garcinia Kola ◽  
Significant Difference ◽  
Anti Inflammatory

Background: Pneumonia is an acute or chronic inflammatory disorder of the lungs, affecting the mucosalareas of the lung.It can becaused by bacteria, viruses or fungi. In some cases, it may be caused by physical or chemical irritants. Kolaviron, a natural bioflavonoid extract from Garcinia kola seeds, has been shown to possess anti-inflammatory properties in Flu-like conditions which are associated with cough. There has been paucity of information on the likelihood of the effectiveness of kolaviron against pneumonia-infections. Objective: To evaluate the anti-bacterial and anti-inflammatory effects of kolaviron on albino Wister ratsinduced with pneumonia using Klebsiella pneumonia. Methods: Powdered Garcinia kola seeds were extracted, withn-hexane and 100% methanol as solvents, using Soxhlet extractor. Astandard method was used to obtain kolaviron from the seed extracts. A total of 24 albino wistar rats were randomly divided into six groups A to F,each comprised four rats. The rats were allowed for 1 hour to acclimatize in very cold environments using ice packs. A standardized 1.0 x10-5 mg/ml culture suspension was intranasally inoculated to the rats for 10 days to induce pneumonia-like symptoms. Thereafter, the kolaviron was administered to the rats such that a500mg/kg kolaviron extract was given once daily to groups A (male rat) and B (female rat). Groups C (male rat)and D (female rat)received 250mg/kg of kolaviron extract once daily while group E rats were given 0.5 ml of dimethyl sulfoxide (DMSO) once daily, and served as negative control. The rats in Group F received 2.86 mg/kg of ofloxacin once daily, and served as positive control. All the treatments were done for a period of 5 days.Then 10 days after thetreatments, the animals were sacrificed and the lungs were harvested for hydrostatic lung test and histopathological examination. An overnight broth culture of Klebsiella pneumonia was streaked in sterile molten nutrient agar maintained at 37o C for 24hrs.Later a stock of 500mg/ml of kolaviron was prepared in DMSO. Two–fold dilutions were performed to obtain the following concentrations with the stock inclusive: 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, and 1.565%.The anti-Klebsiella pneumonia activity of the kolaviron extract was determined using agar well diffusion methodsand incubation was done at 37o C for 24 hrs.Student t-test and One-way Analysis of variance (ANOVA) were used for comparison of mean differences between and among groups. Results: The sensitivity of Klebsiella pneumonia to kolaviron was concentration-dependent. There was increase in antiKlebsiella pneumonia activity with decrease in kolaviron concentration. Kolaviron (KV), at 500mg/kg concentration, was efficacious and showed significant anti-inflammatory effects, (P<0.0001). This was also confirmed in the histopathological examinations. The 3.125% concentration of the kolaviron gave IZDs that ranges from 25.68±3.33 mm in day 1 to 27.33±2.78 mm in day 5. Treatment with kolaviron was sex-dependent having significant difference,(p<0.0001), when pretreatment and post-treatment effects were compared between male and female rats. Conclusion: Kolaviron can be used as an agent in the treatment of pneumonia as the KV possesses anti-inflammatory and anti-Klebsiella pneumonia activities.

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