ß-Sitosterol Alters the Inflammatory Response in CLP Rat Model of Sepsis by Modulation of NFκB Signaling.

PURPOSE: Sepsis originates from the host inflammatory response, especially to bacterial infections, and is considered one of the main causes of death in intensive care units. Various agents have been developed to inhibit mediators of the inflammatory response; one prospective agent is ß-sitosterol (ßS), a phytosterol with a structure similar to cholesterol. This study is aimed at evaluating the effects of ßS on the biomarkers of inflammation and liver function in cecal ligation and puncture- (CLP-) induced septic rats. METHODS: Thirty male Wistar rats were divided equally into six groups as follows: sham, CLP, CLP+dexamethasone (DX, 0.2 mg/kg), CLP+ßS (1 mg/kg), CLP+imipenem (IMI, 20 mg/kg), and CLP+IMI (20 mg/kg)+ßS (1 mg/kg). Serum levels of IL-1ß, IL-6, IL-10, AST, ALT, and liver glutathione (GSH) were assessed by ELISA. Liver expression levels of TNF-α and NF-κBi mRNAs were evaluated by RT-qPCR. RESULTS: Serum concentrations of IL-1ß, IL-6, IL-10, ALT, and AST and mRNA levels of TNF-α and NF-κBi were all significantly higher in septic rats than in normal rats (p < 0.05). Liver GSH content was markedly lower in the CLP group than that in the sham group. ßS-treated rats had remarkably lower levels of IL-1ß, IL-6, IL-10, TNF-α, NF-κBi, AST, and ALT (51.79%, 62.63%, 41.46%, 54.35%, 94.37%, 95.30%, 34.87%, and 46.53% lower, respectively) and greater liver GSH content (35.71% greater) compared to the CLP group (p < 0.05). CONCLUSION: ßS may play a protective role in the septic process by mitigating inflammation. This effect is at least partly mediated by inhibition of the NF-κB signaling pathway. Thus, ßS can be considered as a supplementary treatment in septic patients.

Ficus Carica L. Latex: Possible Chemo-Preventive, Apoptotic Activity and Safety Assessment.

Hepatocellular carcinoma is the third cause of cancer-related mortality with the low 5-year survival in which more than 50 percent of patients have recurrent cancer within 2 years of treatment. The present study investigated the cytotoxicity and lethal dose of Ficus carica L. (Figure) latex and phytochemical composition of effective fraction. Figure latex was collected in summer and 4 fractions of Figure latex were prepared. The cytotoxic effect of each fraction was studied and the most effective fraction was selected for apoptosis assay, acute toxicity study, and phytochemical analysis using column chromatography. The isolated compounds were identified by 1H-NMR, 13C-NMR, and mass spectroscopy. Chloroform fraction was the most effective fraction with the IC50 value of 0.219 and 0.748 mg/mL for HepG2 and NIH cell lines, respectively. Presence of cells in early apoptotic phase was documented by flow cytometry assay. Single dose administration of 2g/kg of fraction did not cause any death. Phytochemical analyses confirmed presence of lupeol acetate and lupeol palmitate in chloroform fraction. The present study revealed that the chloroform fraction is not only 3.4 times more toxic in HepG2 cell line but also has low in-vivo toxicity which could be considered as a good candidate for a chemo-preventive agent.

Monomethyl fumarate alleviates sepsis-induced hepatic dysfunction by regulating TLR-4/NF-κB signalling pathway.

AIMS: Sepsis is a potentially fatal illness that can lead to impairment of multiple organs such as liver. The condition is deeply associated with oxidative stress and inflammation. Monomethyl fumarate (MMF) has manifested antioxidant and immunomodulatory properties. The aim of current study was to evaluate protective effects of MMF in sepsis-induced hepatic dysfunction. MAIN METHODS: Sepsis was induced by cecal ligation and puncture (CLP). Wistar rats were assigned to one of sham, CLP, CLP + dexamethasone (as positive control of inflammation) and CLP + MMF groups. Levels of serum IL-1ß, IL-6, IL-10, AST, ALT and γ­GT were quantified. Furthermore, Hepatic levels of GSH and MDA and mRNA expression of TNF and NFKBIA along with hepatic protein level of TLR-4 were assessed. Also, histopathological study of liver was carried out to evaluate hepatic injuries. KEY FINDINGS: Septic rats demonstrated risen levels of IL-1ß, IL-6, IL-10, AST, ALT and γ­GT, while treatment with dexamethasone or MMF attenuated these levels. Moreover, enhancements in protein level of TLR-4 and mRNA levels of TNF and NFKBIA were observed in CLP rats. These elevations were mitigated in CLP-induced rats that were treated with either dexamethasone or MMF. Treatment with dexamethasone or MMF also shifted sepsis-induced disturbance in the levels of GSH and MDA towards sham levels. Hepato-protective effects of dexamethasone and MMF were further confirmed by histopathological observations. SIGNIFICANCE: Our findings imply that MMF alleviates sepsis-induced hepatic dysfunction by mitigating the inflammatory and oxidative state and this effect is at least partly mediated by the inhibition of TLR-4/NF-κB signalling pathway.

Diazinon Interrupts Ovarian Steroidogenic Acute Regulatory (StAR) Gene Transcription in Gonadotropin-Stimulated Rat Model.

Organophosphate pesticides are considered as endocrine disruptors that interfere with reproductive functions. The corpus luteum (CL) is a transient endocrine gland that produces progesterone, a crucial hormone for a successful beginning and maintenance of pregnancy. Steroidogenic acute regulatory protein (StAR) facilitates the rate-limiting transfer of cholesterol from the outer mitochondrial membrane to the inner organelle membranes. We investigated the effect of Diazinon (DZN), an organophosphate, on StAR mRNA expression by Sybergreen Real Time-PCR in a time-dependent manner in luteal phase. Fifty immature female Wistar rats (24-day-old) were injected with a single injection of Pregnant mare's Serum Gonadotropin (PMSG) followed by a single injection of human Chorionic Gonadotropin (hCG), 48 h later. Then, DZN was administered in a single dose (70 mg/kg bw, I.P), controls received only the vehicle, 12 h post-hCG injection. Ovaries were collected in 4 h. intervals from 8 to 24 h post-hCG injection. Then, hCG stimulation transcript levels of StAR gene were significantly altered in the hormone-stimulated rats following DZN treatment. In addition, histological study showed that the CL diameter in DZN-treated group was smaller than control group (p = 0.000). Our findings suggest that the critical step in the function of CL is disrupted by DZN and may correlates with female reproductive damage.

Ethanol Extract of Crataegus Oxyacantha L. Ameliorate Dietary Non-Alcoholic Fatty Liver Disease in Rat.

BACKGROUND: Non-alcoholic fatty liver (NAFLD) is one the most prevalent disease worldwide which characterized by fat accumulation in liver with no established efficient therapy. We designed this study to investigate protective and therapeutic effect of Crataegus oxyacantha L. (C. oxyacantha) on NAFLD induced by high fat diet in rat models. METHODS: NAFLD was induced by High Fat Diet+fructose (HFD), 45 Wistar rats were divided to 8 groups including control, HFD, HFD+diet change, HFD+diet change+C. oxyacantha 20 mg/kg, co treatment of HFD+C. oxyacantha 10, 20 and 40 mg/kg, and normal diet+C. oxyacantha 40 mg. C. oxyacantha was administered orally. Effectiveness of the C. oxyacantha was assessed through measuring the biochemical factors, and oxidative stress marker (FRAP, GSH, and MDA). Histopathological study was performed using H & E staining. RESULTS: The diet change from high fat to low fat ameliorated liver damage. However, consumption of C. oxyacantha (10 & 20 mg/kg) caused significant reduction in the level of all examined liver biomarkers specially LDH, that showed C. oxyacantha can restore the hepatocyte damage due to HFD. The C. oxyacantha showed a protective effect which was more prominent in the animals treated with the 20 mg/kg C. oxyacantha. The administration of C. oxyacantha caused increased antioxidant status (GSH and FRAP levels) and decreased lipid peroxidation in treated animals. MAJOR CONCLUSION: Accordingly, C. oxyacantha have both therapeutic and protective effect for NAFLD and may be a potential candidate for further assessments in clinical studies.

Different Genotype of rs3130932 Single Nucleotide Polymorphism Between Gastric Cancer Patients and Normal Subjects.

PURPOSE: Octamer binding transcription factor B gene (OCT4) is responsible for development and self-renewal maintenance of embryonic stem cells. The rs3130932 single nucleotide polymorphism (SNP) may play a role in tumor genesis. Because of high prevalence of gastric cancer in north of Iran, this study was investigated role of rs3130932 polymorphism and stomach cancer. METHODS: Blood samples were collected from 100 informed gastric cancer patients and 100 age and sex-matched healthy individuals, and were genotyped for the presence of rs3130932G allele by ssp PCR. RESULTS: The mean age of participant (n = 200) was 67.83 ± 10.878 years. In genotyping and allelic analysis, TG genotype increased 66.147 times more likely to develop stomach cancer than the TT genotype, and disease risk increases 140.496 times more in GG genotype in comparison with TT genotype. CONCLUSION: This study clearly emphasis on different genetic profile in this population and show that the rs3130932G allele and odds of gastric cancer are related to each other in northern of Iran.

Epidermal growth factor receptor gene polymorphisms and gastric cancer in Iran.

BACKGROUND: Epidermal growth factor receptor (EGFR) is a transmembrane receptor which contributes to many processes involved in cell survival, proliferation and inhibits apoptosis, that may lead to cancer development. Gastric cancer is one of the most common diseases of digestive system that has low 5-year-survival. The aim of this research was to determine the significance of EGFR tyrosine kinase domain gene polymorphisms in gastric cancer in Iran. MATERIALS AND METHODS: In the present study, 83 patients with gastric cancer and 40 normal subjects were investigated for EGFR gene polymorphisms in exons 18-21 by PCR-SSCP. Then, DNA sequencing was conducted for different mobility shift bands. Finally the data were statistically analyzed using the chi-2 test and the SPSSver.16 program. RESULTS: Exon 18 of EGFR gene showed three different bands in SSCP pattern and DNA sequencing displayed one mutation. SSCP pattern of Exons 19 and 21 did not show different migration bands. Exon 20 of EGFR gene revealed multiple migrate bands in SSCP pattern. DNA sequencing displayed 2 mutations in this exon: one mutation was caused amino acid change and another mutation was silent. CONCLUSION: It may be that EGFR tyrosine kinase gene polymorphisms differ between populations and screening could be useful in gastric cancer patients who might benefit from tyrosine kinase inhibitor therapy.