مجله سلول و بافت
سال ششم شماره 2 (تابستان 1394)

The aim of this study was to investigation cobalt nano-particles effects on expression level of key genes involved in sesamin biosynthetic pathway including CYP81Q1، CYP81Q2، CYP81Q3 and C3H in Sesamum indicum L. cell suspension culture. First sesame cell suspension culture was established. For this purpose، Karaj1 hypocotyle cultivar، 0. 6 mgL-1 BAP، 3 mgL-1 NAA were used. Different concentrations of cobalt nano-particles elicitor including 0. 25، 0. 5 and 1 mgL-1 were added to the 18 days-old culture and sampling was carried out after 2، 8 and 24 hours post treatment. Expression level of mentioned genes was measured by the Real-Time qRT-PCR technique. Data analysis and figures depiction were done using Excel. Results showed that MS medium consisting of 0. 6 mgL-1 BAP، 3 mgL-1 NAA and 30 gL-1 socrose، 26ºC، 130rpm and darkness condition are optimum situation for condensed sesame suspension production. Adding cobalt nano-particles to the suspension culture، led to increasing genes expression level، especially، at 0. 5 mgL-1 elicitor concentration in different times. Since this elicitor had no negative effects on the mentioned genes expression، the effect of higher concentrations on these genes expression can be studied in the future. Negative effects of this elicitor on the expression of genes involved in sesamin biosynthesis means decreasing sesamin content. Based on obtained results elicitation of sesame cell suspension culture by cobalt nano-particles led to the increasing genes expression involved in the sesamin biosynthetic pathway. Therefore، this elicitor can be used to induce higher expression levels of these genes.

The aim of this study was to evaluate the negative effects of different doses of silvernanoparticles on fertility potential، chromatin structureand DNA integrity of epididymal sperm in mice. Twenty four male mice were divided into 4 groups (n=6) including one control and three experimental groups for orally administration of silver nanoparticles with three different doses (50، 100 and 200 μl/kg/day) for 5 weeks. Then، cauda epididymal spermatozoa were aspirated for analysis of sperm parameters using general microscopy methods and according to WHO criteria. Chromatin condensation، intensity of sperm chromatin abnormalities and protamine deficiency were assessed by three different cytochemical tests including aniline blue (AB)، toluidine blue (TB) and chromomycin A3 (CMA3) tests respectively. Results showed that third group (treated with the highest silvernanoparticles dose)، had the lowest sperm number، the lowest of sperm fast motility percentage and the lowest sperm percentage having normal morphology significantly in comparison with control and others groups. The II and III groups significantly showed differences in sperm chromatin condensation and deficiency comparing to other groups. In this study the negative effect of silver nanoparticles on mice sperm parameters، sperm chromatin structure and DNA integrity were observed. It is believed that the negative effects of silver nanoparticles on sperm quality are significant and depend on used doses.

In this study zinc oxide nanoparticles (ZnO nanoparticles) toxicity effects on some male sexual hormone levels in blood serum and testicular tissue in adult intact rats was studied. The rats were divided into a control and four treatment groups. ZnO nanoparticles at concentrations of 2. 5 and 1 g/kg were orally administered for two treated groups and intra-peritoneal were synchronic used for two other treated groups. Nanoparticles effect and its damages on testicular tissue were studied one، seven and fourteen days after treatment. Blood serum tests showed that serum LH and FSH levels increased in zinc oxide nanoparticles treated groups in comparison with control (P<0. 05). Also، testes tissue slides examination showed specific tissue damages in all four zinc oxide nanoparticles treated groups in comparing to control. These injuries represent irregularities in the cell layers، cell-free seminiferous tubules، congestion in seminiferous tubules area and changes in spermatogene cell nucleus. Results of the present study show nanoparticle zinc oxide effect on morphological structure and sex hormones.

In this study، sodium nitroprusside effect as a producer of NO on growth، antioxidant enzyme activities and some physiological parameters in in vitro cultured Melissa officinalis seedlings were studied. After establishment of Melissa officinalis seedling culture، seedlings were treated with different concentrations of sodium nitroprusside (0، 25، 50 and 100 µM). Then، they were harvested for growth، protein content، secondary metabolites، lipid peroxidation level and antioxidant enzymes activities analysis after 10 days. Results showed that growth and protein content decreased with increasing concentration of sodium nitroprusside. Phenolic compounds amount، total flavonoids and anthocyanins increased by sodium nitroprusside. Also Malondialdehyde content as lipid peroxidation product was significantly increased. In response to oxidative stress، chatalase and peroxidase enzymes activities were respectively increased at 50 and 25 µM sodium nitroprusside while enzymes activities showed decreasing at 100 µM. Overall، the results indicate that sodium nitroprusside as NO producer damage the cell membrane. Therefore reduced Melissa officinalis seedling growth and caused oxidative stress in them. To overcome produced free radical، secondary metabolites amounts and antioxidant enzymes activities were increased.

In this study the synergic effect of Chiton lamyi foot alcoholic extract and decellulation rat brain’s tissue on angiogenesis in hen (chick?) chorioalantoic membrane (CAM) was investigated. In this study، 70 fertilized eggs of Ross strain were divided into 7 treatments groups in ten repetitions: 1. control، 2. DMSO، 3. decellulation rat brain’s tissue، 4. 10mgμl-1 Chiton lamyi foot alcoholic extract، 5. 20mgμl-1 Chiton lamyi foot alcoholic extract، 6. decellulation rat brain’s tissue & 10mgμl-1 Chiton lamyi foot alcoholic extract and 7. decellulation rat brain’s tissue & 20mgμl-1 Chiton lamyi foot alcoholic extract. All of eggs were placed in the incubator for eight days and treatments induction was done in 8th day. Then they photographed using photo steriomicroscope in 12th day. Blood vessels branches length and number were evaluated by Image J software in treatment sites on chorioalantoic membrane. Data were analyzed using ANOVA، SPSS Test (P<0. 05). Results showed that Chiton lamyi foot alcoholic extract، 10 and 20mgμl-1 Chiton lamyi foot alcoholic extracts with decellulation rat brain’s tissue treatments significantly decreased blood vessels branches length and number in comprising with control and DMSO treatments (P< 0. 05). The Synergic usage of Chiton lamyi foot alcoholic extract and decellulation (acellular) rat brain’s tissue has dose inhibitory effect on hen chorioalantoic membrane (CAM) angiogenesis.

The purpose of this study was to evaluate the effects of sub-lethal osmotic stress (325، 350، 375 and 400 mOsm) during cryopreservation in commercially diluter medium (Bioexcell) on Holstein mail cow sperm qualitative characters after freezing-thawing. Semen was collected from four Holstein mail cows using artificial vagina two times for a week. All of obtained semen mixed together and then were divided into five equal parts. Each part was freeze-melted according to the experimental treatments consisting of different osmotic stresses. 300 mOsm treatment medium was applied as control group. Sperms motility and progressive motility were assessed by computer assessment semen analysis. Also sperms viability، membrane integrity، mitochondria activity and membrane lipid peroxidation of frozen-thawed semen were assessed using Eosin-Nigrosin، hypo osmotic swelling test، Hankok، Rhodamin 123 and TBA procedures، respectively. Results showed that 375 mOsm osmotic stress treatment had the most significant improvement of motility%، progressive motility and viability in comparison with other treatments. Also mitochondria activity in 350 and 375 mOsm osmotic pressures treatments was significantly higher than other experimental treatments (P<0. 05). Different osmotic stresses treatments did not show significant effect on sperm morphology and membrane lipid proxidation. It is seemed that mild and sub-lethal osmotic stresses induction in cow sperm freezing diluters can significantly improved some of sperm qualitative characters such as their motility and viability.