Effects of pentoxifylline on the histological and ultrastructural features of vitrified mouse ovarian tissue

Background: Vitrification is a process that can be used to preserve gonads in the healthy and natural status. Oxidative stress is one of the disadvantages of vitrification. Pentoxifylline [PTX] is an antioxidant that can reduce reactive

oxidative stress effects

Objective: We aimed to investigate the effects of PTX on histological and ultrastructural features of vitrified and non-vitrified mouse ovarian tissue

Materials and Methods: Twenty-five adult female Balb-C mice were randomly and equally divided into control group: the ovaries did not receive any treatment; experimental 1 and 2: the vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin without and with PTX, respectively, for 30 min; sham 1 and 2: the non-vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin and were incubated without and with PTX, respectively for 30 min. The right and left ovaries in all of the groups were evaluated using light and transmission electron microscopy, respectively

Results: The histological and ultra-structural features of vitrified ovaries were seriously damaged. There was non-uniformed germinal epithelium and tunica albuginea, degenerated granulosa cells and stromal cells, puffy basement membrane and irregular thickness of zona pellucida, as well as a pyknotic nucleus and bubbly and segmented ooplasmic in the follicles. Also, ovarian tissues were damaged by the PTX in the non-vitrified ovaries

Conclusion: Vitrification can damage the histological and ultra-structural features of the ovary in mouse models. PTX as an antioxidant, with concentration of 1.8 mM could not prevent and restore these damages and had no adequate effects on the vitrified ovarian tissues

Protective effects of curcumin co-treatment in rats with establishing chronic variable stress on testis and reproductive hormones

Background: Protracted and repeated exposure to chronic variable stress [CVS] may lead to reproductive dysfunction. It is a basic cause of male infertility. Curcumin [CUR] is an active fraction of turmeric that used in traditional Chinese medicine. CUR represents various pharmacological activities

Objective: The purpose of this study was to determining the effects of CUR on testis and testosterone, follicle stimulating hormone [FSH] and luteinizing hormone [LH] in rats with establishing chronic variable stress

Materials and Methods: Twenty-one adult male Sprague-Dawley rats were divided into three groups: 1] control, 2] CVS and 3] CVS+ CUR [100 mg/kg/day dissolved in 0.5 mL of olive oil]. All of the animals in control, CVS, and CVS+CUR groups were sacrificed after 15 days. Testosterone, FSH, LH, and testis damage were evaluated

Results: Significant changes in the normal range of testosterone, FSH, LH serum levels and seminiferous tubule apoptotic cells were detected in CVS group compared to the control rats [p=0.02]. These parameters changed to a less extent in CVS+CUR animals compared to the CVS rats [p=0.02]

Conclusion: Our findings propose that curcumin might have curative potential on the reproductive system function and its impairment. It's regulated by stress and reproductive-related hormones

Lectin histochemistry showed a heterogeneous population of cells among human mesenchymal stem cells isolated from adipose tissue

Objectives: Adipose tissue as an appropriate source of Mesenchymal Stem Cells [MSCs] has the potential to differentiate into multiple lineages. Glycoconjugates content of the MSCs can be considered as biomarkers in self-renewal, pluripotency and differentiation processes. In this study, the lectin profile of MSCs isolated from adipose tissue was detected and according to that, a subpopulation was determined

Materials and Methods: MSCs were isolated from adipose tissue by explanting of the tissue pieces. The FITC-conjugated lectins, WGA, UEA, PNA, BSA and PWM were used to detect the terminal sugar residues. The cells were then counterstained with DAPI. The intensity of the reaction was evaluated by ImageJ software. The cells were also stained with PAS method

Results: MSCs were reacted with all lectins with different intensity of the reactions. The cells reacted with WGA, UEA, and BSA "strongly" and with PWM "moderately" and with PNA with "weak" intensity. The morphological analysis of the isolated MSCs revealed the existence of the two different cell types in the cultures. Two types of cells were detected according to nucleus size and lectin reactivity. The cells with large nuclei constitute 20.62% of the total cells and stained significant more intensity by UEA and less intense with PWM [both P=0.014] and PNA [P=0.044]. Flow cytometry with CD34 shows that these large cells were not endothelial cells

Conclusion: The MSCs derived from adipose tissue seem to be a heterogeneous populations and lectin profile of the cells showed that they are different in the expression of the glycoconjugates

Effects of Anethum graveolens L. [dill] on oocyte and fertility of adult female rats

Our previous studies revealed Anethum graveolens L. caused some changes in female reproductive system that induced infertility. Therefore, in this study, oocyte changes as one of probable reasons of infertility were investigated. In this study, 59 adult female rats were divided into 3 groups of control, low dose [0.5 g/kg] and high dose [5 g/kg] of dill seed aqueous extract [LDE and HDE] treated groups that were gavaged with 1 ml of each dose for 10 days [2 estrous cycles]. Vaginal smears were prepared daily. Oocytes of superovulated animals were extracted and their morphometrical changes were measured [n=5]. Oocyte cell membrane glycoconjugates were stained with UEA, PNA, and DBA-FITC lectins [n=5]. Ultrastructural studies of oocytes were performed using TEM [n=5]. The number, weight, and crown-rump length of newborns were examined in three groups after mating with untreated males [n=5]. Data were analyzed using SPSS software. Results demonstrated that the duration of the estrous cycle, the diestrus phase and progesterone concentration in the experimental groups increased significantly compared to the control group [p<0.05]. Granulosa cells of corpus luteum in HDE-treated group were larger and clearer. The intensity reactions of galactose/N-acetylgalactoseamine terminal sugar of oocyte decreased insignificantly in experimental groups compared to the control group p>0.05. Duration of mating to pregnancy increased and the weight and crown-rump length of newborns decreased in experimental groups significantly [p<0.05]. Dill seed aqueous extract can induce infertility without any effect on oocyte structure

Effect of follicular fluid and platelet-activating factor on lactate dehydrogenase C expression in human asthenozoospermic samples

Background: Application of follicular fluid [FF] and plateletactivating factor [PAF] in artificial insemination improves sperm motility. Lactate dehydrogenase C [LDH-C] is a key enzyme for sperm motility. In this study, the effects of FF and PAF on the sperm motility index and LDH-C expression were investigated. Moreover, LDH-C expression was compared between asthenozoospermic and normozoospermic samples

Methods: The expression of LDH-C was examined by quantitative real-time polymerase chain reaction [q-RT PCR] and western blotting after it was treated with optimized concentrations of FF and PAF in twenty asthenozoospermic samples. Also, LDH-C expression was evaluated in five normozoospermic samples

Results: Samples with 75% FF and 100 nM of PAF had an increase in their percentages of progressive and slowly motile sperms and a decrease in their percentages of non-progressive and non-motile sperms. Moreover, LDH-C mRNA transcripts were not changed following PAF and FF treatment, and LDH-C protein was detected in highly progressive motile specimens treated with FF in the asthenozoospermic samples. Furthermore, LDH-C expression was more detectable in the normal sperms

Conclusion: Our results indicated that PAF had more beneficial effects than FF on sperm motility in the asthenozoospermic samples [P=0.0001], although the LDH-C expressions of the sperms were not changed significantly in both groups. We found no association between LDH-C expression and sperm motility after FF and PAF actions. This finding, however, requires further investigation. The fact that LDH-C protein was detected in the normozoospermic, but not asthenozoospermic, samples could be cited as a reason for the infertility in these patients

Effects of L-Carnitine and Pentoxifylline on carbohydrate distribution of mouse testicular sperm membrane

Background: The glycoconjugate content of sperms indicates their physiological and fertility properties. Lectin reactivity is indicative of intact, capacitated, and acrosome-reacted sperms. In the epididymis, sperms experience maturation, glycoconjugate modification, and simultaneously, higher L-carnitine [LC] concentrations. The aim of this project was to evaluate the effects of LC and Pentoxifylline [PF] on the integrity, capacitation, and acrosomal reaction of sperms by studying their lectin reactivity

Methods: Mouse testicular sperm samples were divided into three parts. Each sample was added Ham's F10 [control] or media containing 1.76 mM LC or PF. At 30 and 90 minutes after incubation, sperm motility was assessed. Peanut agglutinin [PNA], wheat germ agglutinin [WGA], and Concanavalin A [Con A] were used to detect non-acrosome-reacted, non-capacitated, and acrosome-reacted sperms, respectively and the frequency was evaluated by flow cytometry. Statistical analysis was performed using the ANOVA

Results: Sperm motility increased after 30 and 90 minutes of incubation in the LC- and PF-treated cultures [P=0.001]. LC administration created a significant increase in the percentage of the non-acrosome-reacted sperms compared to the control sperms after 30 and 90 minutes [P=0.02 and P=0.03, respectively]. The frequency of the non-capacitated sperms in the LC-treated group increased compared to the control sperms after 30 minutes significantly [P=0.01]

Conclusion: Although the administration of LC and PF enhanced sperm motility, LC also impacted glycoconjugates on the sperm surface. Glycoconjugates are involved in the interaction between the sperm and the zona pellucida and subsequently fertilization, thereby probably influencing the male fertility state

Effects of L-carnitine and pentoxifylline on the activity of lactate dehydrogenase C[4] isozyme and motility of testicular spermatozoa in mice

Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitine. In addition, lactate dehydrogenase C[4] [LDH-C[4]] gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C[4] enzyme activity upon L-carnitine [LC] and Pentoxifylline [PTX] administrations in mice. We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium [control], the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization [WHO] criteria. Sperm LDH-C[4] enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. Sperm motility increased after 30 min of incubation in LC- and PTX-treated group [p<0.001]. LC and PTX administrations showed a significant increase in the LDHC[4] enzyme activity of sperm compared to that of the controls after 30 min [P=0.04 and 0.01, respectively]. The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C[4] enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX

Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality

Sperm cells extracted from testes [TESE] have poor chromatin quality and motility. Various substances are used in the laboratory to increase sperm motility and improve the ART outcomes; however, there are few research which considered improving both sperm motility and chromatin quality. The aim of this investigation was to evaluate the improvement of the testicular sperm motility and chromatin quality exposed to L-carnitine [LC] and L-acetyl-carnitine [LAC], which are normally concentrated in testis and epididymis, compared with Pentoxifylline [PF], which used for sperm motility enhancement in IVF procedures. TESE samples from 30 male mice divided into four parts. The sperm samples were added to Ham' F10 [control] or the media contained 1.76mM of LC, LAC or PF], then, the samples were kept in the room temperature for 30, 90 and 180 min. At each time step, sperm motility and chromatin quality were assessed. Chromatin quality was evaluated by chromomycin A3 and aniline blue. Statistical analysis was performed using one way analysis of variance [ANOVA]. A p-value less than 0.05 were accepted as a statistically significant difference. The results showed LC, LAC and PF significantly increased the sperm motility. However, sperm chromatin quality only improved significantly by administration of LC and LAC. Administration of LC and LAC to the testicular sperm samples can lead to improve both sperm motility and chromatin quality. It may be because they can mimic in vivo sperm condition during late spermatogenesis