Pathobiology Research
Year:2017 | Volume:19 | Issue:4 |Papers Count:7

Objective: There are numerous strategies to prevent hepatotoxicity caused by doxorubicin therapy. These strategies include exercise as well as herbal antioxidants such as curcumin to reduce the toxic effects of doxorubicin. This study aims to evaluate the effects of six weeks of continuous training with and without nanocurcumin supplementation on doxorubicininduced hepatotoxicity in an aging rat model.Methods: We randomly divided 42 Wistar male rats into 7 groups: control saline, control doxorubicin, nanocurcumin+doxorubicin, nanocurcumin+saline, continuous training+ doxorubicin, continuous training+saline, and continuous training+nanocurcumin+ doxorubicin. The rats received intraperitoneal injections of D-galactose (100 mg/kg) to induce ageing. The training groups ran on a treadmill for six weeks, five days per week with a gradual increase from 25 min/day to 54 min/day at a velocity of 15 m/min to 20 m/min. In the last fifteenth days, rats scheduled to received doxorubicin had a cumulative dose of 15 mg/kg of body weight (daily: 1 ml/kg). Nanocurcumin supplement (daily: 100 mg/kg body weight) was administered to the respective groups. Assessment and analysis were conducted after homogenization of the liver tissue biopsy.Results: Doxorubicin caused a significant decrease in glutathione peroxidase and a slight increase in malondialdehyde in the liver. On the other hand, continuous training with doxorubicin treatment prevented the decrease of glutathione peroxidase and increase in malondialdehyde in the liver that was caused by doxorubicin. Also, six weeks of continuous training with nanocurcumin supplementation caused a significant decrease in malondialdehyde and increased glutathione peroxidase in the liver compared to the control doxorubicin group.Conclusion: Based on the results, the combination of nanocurcumin supplementation and continuous training in the doxorubicin-induced aging rat model have led to a precautionary effect and up-regulation of antioxidant defense. Continuous training appeared to have more beneficial effects than nanocurcumin supplementation in reducing doxorubicin-induced hepatotoxicity.

Objective: Atherosclerosis is the most common cause of coronary heart disease and a major cause of ischemic heart diseases. We have investigated the effect of eight weeks of aerobic training on the serum levels of leptin, IL-6, and TNF-α in inactive elderly women.Methods: In this semi-experimental study, participants were selected by convenience sampling. We divided 21 subjects into two groups - active (n=10) and non-active (n=11). The program training included 60 min per session, 3 times per week for 8 weeks of aerobic exercise with 50-70 HRR%. Blood samples were taken before and after 8 weeks of aerobic training. We used the t-test for dependent and independent samples for intra- and intergroup comparisons. For all statistical comparisons, the level of significance was set at P<0.05.Results: The aerobic training groups had a significant reduction in leptin (P=0.001), IL-6 (P=0.01), and TNF-α (P=0.001) after 8 weeks of aerobic training compared with the control group. The intergroup variations in weight, BMI, leptin, and TNF-a had a significant change (P<0.05).Conclusion: The results of the present study suggested that endurance training decreased leptin, IL-6, and TNF-a levels, effectively improved cardiovascular health, and reduced the risk of atherosclerosis.

Objective: Melanoma differentiation association gene-7 (MDA-7) /IL24 is a tumor suppressor gene. The iNGR peptide sequence, with excellent tropism to surface integrins has been employed for targeting of therapeutic molecules toward tumor cells. The purpose of our study was to construct a plasmid expressing modified MDA-7 fused with an iNGR peptide for better targeting to tumor cells.Methods: At first, we amplified the MDA-7 sequence by PCR, while the reverse primer contained the iNGR peptide sequence to add it to the end of a new MDA-7 gene. The resultant MDA7-iNGR and MDA-7 were cloned into a pCDNA3.1 eukaryotic expression vector. The accuracy of cloning methods, integrity of the plasmids, and sequence were sequentially evaluated by digestions, colony-PCR, and sequencing. The expressions of the plasmid constructs were assayed by ELISA following their transfection into Ad-293 cells. Next, the plasmids were transfected into Hep-G2 cells and their mRNA were converted to cDNA. We assessed the gene expression levels of Gadd153 and Bax. As the final step, apoptosis induction of Hep-G2 cells following transfection was evaluated by the help of PI/Annexin V staining according to flow cytometry.Results: The results showed the integrity of construct backbone in addition to reading frame of the MDA-7-iNGR sequence. A suitable expression/secretion of modified the MDA-7.iNGR protein was detected by ELISA assay of the culture supernatant when compared to the control construct that expressed unmodified MDA-7. The viabilty test demonstrated no benefit for this kind of modification of the MDA-7 protein. Real-time PCR and flow cytometry analyses revealed that the addition of iNGR to MDA-7 caused a decrease in its apoptotic effect on hepatic tumor cells compared to the normal protein. The modified protein had significant apoptosis induction compared to the negative control group (P<0.01).Conclusion: Although the new pCDNA/MDA-7.iNGR plasmid expressed iNGR-fused MDA- 7 protein efficiently, it could not improve the natural apoptosis property of normal MDA-7.

Objective: The aim of this study was to evaluate the effect of different concentrations of ovarian tissue extract on isolated preantral follicle growth, maturation, and hormone production of neonatal mouse ovaries after three-dimensional culture.Methods: We obtained the tissue extract from adult mouse ovaries and determined their protein concentrations by the Bradford method. The experimental group comprised mechanically isolated preantral follicles after encapsulation with alginate hydrogel in α-MEM supplemented with different concentrations of ovarian tissue extract and in 10% fetal bovine serum as the control group. Both groups were cultured for 12 days. At the end of the culture period, we assessed the mean diameters, survival and maturation rates, production of 17-β estradiol, and progesterone hormones.Results: The follicles cultured in different concentrations of ovarian tissue extract underwent degeneration, with the exception of the group that contained tissue extract (onehalf the protein level of FBS) which had a 71.78% survival compared to the control (75%). The average follicle diameter, rate of MII oocytes, production of 17-β estradiol, and progesterone hormones were significantly higher in the group supplemented with ovarian tissue extract (one-half the protein level of FBS) compared to the control group (P<0.05).Conclusion: Ovarian tissue extract, as a proliferation and maturation factor, leads to growth, maturation, and better function of preantral follicles. This extract can be used to improve the in vitro follicular culture in a dose-dependent manner.

Objective: Fetal hemoglobin is the predominant hemoglobin expressed by gamma globin. However, in adults, fetal hemoglobin normally reduces to very low levels of the total hemoglobin. The increase in levels of fetal hemoglobin can ameliorate the severity of β-hemoglobin disorders such as sickle cell disease and beta-thalassemia. Currently, drugs that have been used for induction of Fetal hemoglobin (HbF) have short-term effects. Dendrosomal nano-curcumin (DNC), which has high solubility and absorption, is able to detect different targets in the cell and affect gene expression. LSD1 is one of the most important gamma globin inhibitors. In this study, we examine the capability of DNC to inhibit the expression of LSD1, GATA1, and FOG1 as well as the increase in gamma globin expression.Methods: We used the K562 cell line for the MTT assay and treatment by DNC. Then the effect of DNC on the increase in expression level ofγ-globin, decrease expression level of LSD1 and transcription factors, GATA1 and FOG1was investigated by Real time PCR.Results: Data acquired from gene expression assays indicated that DNC induced gamma globin expression and decreased expressions of LSD1, GATA1, and FOG1 in a time and dose-dependent manner.Conclusion: Inhibition of LSD1, GATA1, and FOG1 expressions via DNC led to increased gamma globin expression. These results showed that DNC could be a promising treatment for beta-thalassemia and sickle cell disorders, and possibly reduce the severity of symptoms of these patients through the induction of fetal hemoglobin.

Objective: More than 99% of cervical cancers contain human papillomavirus (HPV), particularly the high-risk HPV type 16 (HPV-16). Among therapeutic HPV vaccines, DNA vaccines have emerged as a potentially promising approach. The main problem with DNA vaccination is the efficient delivery of the genes. A different delivery system has been used to bypass this problem. Archaeosomes have shown high stability during oxidative stress. In this study, we prepared the archaeosome Halobacterium salinarum polar lipid and used it as a delivery system and adjuvants for formulation with the E6/E7/L1 chimeric plasmid as an HPV vaccine candidate.Methods: The recombinant pIRES2-plasmid that contained an E6/E7/L1 chimeric gene of HPV were purified after extraction. Halobacterium salinarum total polar lipids were prepared according to a method by Bligh and Dyer. The archaeosome-pDNA complex was prepared by the addition of plasmid DNA to an archaeal lipid solution and the mixture kept at room temperature to allow for complex formation. Particle sizes and zeta potential of the samples were measured using dynamic light scattering. We measured the relative tumor volume after administration of TC-1 cells to C57BL/6 mice.Results: Zeta potential of the anionic archaeosomes was -6.84mV while archaeosome-pDNA complexes were -29 mV. The highly negative zeta potential of archaeosome-pDNA complexes demonstrated excellent loading of the plasmid on the nanoparticle surface and electrostatic stability. The results showed that the archaeosome-containing E6/E7/L1 chimeric gene significantly inhibited the rate of tumor growth in comparison with the control groups.Conclusion: Archaeosomes are easy and cost-economic to prepare and highly stable. They may hold tremendous promise as vaccine delivery vehicles.