@2024 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2015;21(1):21-29

Background
… [1-11] Medicinal plants having antioxidant compounds can improve the sperm and testicular tissue by reducing free radical damage caused by diabetes [12]. Some compounds have been identified in the extract of Launaea acanthodes (Boiss) O. Kuntze from the Asteraceae family, such as flavonoids, terpenoides, saponins, alkaloids, tannins, polysaccharides and monosaccharides such as arabinose, mannose and derivatives of acid glucuronic [13, 14]. Since flavonoids as antioxidant are able to reduce the level of free radical cells, they can be used to reduce the devastating effects of diabetes [15].
Previous Studies
So far, no study has been conducted with respect to the effect of intra-peritoneal injection of aqueous extract of Launaea acenthodes on improving sperm damage caused by diabetes.
Aim(s)
The aim of this study was to investigate the effect of the aqueous extract of Launaea acenthodes on sperm parameters and textual changes of testis in diabetic Wister rats.
Research type
This is an experimental study.
Research society, place and time
Male Wistar rats aged 20 weeks and weighing approximately 148 ± 16 g in animal research laboratory, Department of Biology, University of Payam E Noor, Iran were studied in 2014.
Sampling method and number
27 rats were studied.
Used devices&materials
Animals were kept at approximately 23-25° C, 30-35%of relative humidity and 12-hour light/darkness cycle. Rats were randomly divided into 3 groups of 9 rats in healthy control group, 9 in the control group of diabetic rats and 9 in the treated group with the extract of Launaea acenthodes. Air organ of the Launaea acenthodes plant from 35 to 36 north degrees longitude and 58 to 59 north degree latitude were collected over an area of about 7 hectares. Minimum and maximum altitudes are 1258 and 1482 meters from the sea level, respectively. Soxhlet extraction was carried out. Pure extract, after the removing the solvent and drying, was injected with 300 mg/kg concentration. Experimental model of diabetes (insulin-dependent diabetes) was done in rats with a single intra-peritoneal injection of Alloxan monohydrate (Sigma-Aldrich; Germany) with the amount of 360 mg/kg. Water was used as a solvent of alloxan. Alloxan injection to diabetic control group and treated diabetic group was done with the extract. Blood glucose was measured by glucometer (EasyGluco; Korea) and blood glucose above 300 mg/dl was considered as an indicator of being diabetic. Blood glucose levels over 300 mg/ dl was considered as the indication of diabetes in rats on the first day of the experiment and then all injections was conducted for a month, every other day [16]. … [17] In all groups on day 30 of the experiment, the rats were killed by inhalation of diethyl ether (Merck; Germany) in a closed environment and their testis were extracted for investigating sperm parameters. The progressive movements of the sperms were evaluated as rapid progressive movement, slow progressive movement, and lack of mobility. Hemocytometer Neubauer slide was used to study sperms. Sperm morphology was classified as follows: 1) Normal: sperms which did not have any appearance problems in the head, neck and tail; 2) Abnormal head shapes: sperms which had smaller or bigger head size comparing with normal head size or had heads which were separated from the other parts; 3) Abnormal forms of the neck: sperms which had angle between head and neck or the sperms which had swelling in their neck; 4) Tail abnormal forms: sperms having shorter tail comparing to the normal tail or which had abnormal tails. Measurement of the average of the seminiferous diameter and the average thickness of the epithelium were investigated in the images provided by the software Image J 2 [18]. Statistical analysis of obtained data was analyzed by SPSS 20 software and the Kruskal-Wallis nonparametric analysis of variance. To compare the two groups, Dunn's post hoc test was used.
Findings by Text
The mean percentage of sperm with fast and slow movements in the diabetic control group was significantly decreased compared with the healthy control group. The mean percentage of sperm with no mobility in this group significantly increased compared with the healthy control group. The mean percentage of sperm with fast and slow movements in the experimental group treated with the extract at a concentration of 300 mg of aqueous plant of Launaea acenthodes decreased as compared to control group. This reduction was significant in fast motion, but not in slow motion. The mean percentage of sperm with fast and slow movements in the experimental group treated with the extract was significantly increased compared to the diabetic control group. Also, the mean percentage of sperms with no mobility in the experimental group treated with extract of Launaea acenthodes significantly increased compared with the healthy control group, but significantly decreased compared to the diabetic control group. The percentage of normal sperms in the diabetic control group was significantly decreased compared to the healthy control group. Also, the mean percentage of normal sperms in the experimental group treated with aqueous extract showed significant decrease compared with the healthy control group, but it was increased significantly compared to the diabetic control group. The percentage of abnormal forms of head, neck and tail of the sperm in the diabetic control group significantly increased compared with the healthy control group. The percentage of abnormal forms of head, neck and tail of the sperm in the experimental group treated with the extract, also, increased compared with the healthy control group. This increase was significant for abnormal forms of head and tail, but it was not significant for the abnormal forms of neck. The percentage of abnormal forms of head, neck and tail of the sperms in the experimental group treated with the extract showed significant decrease compared to the diabetic control group (Table 1; Fig. 1). The sperm count (x106) in the diabetic control group was significantly decreased compared with the healthy control group. Also, the sperm count in the experimental group treated with the extract, decreased significantly compared to the healthy control group, but it was significantly increased compared to the diabetic control group (table 1). The weight of testis, testis length, testis width, seminiferous tubules diameter and thickness of the epithelium of seminiferous tubules in the control group significantly decreased compared with the healthy control group. The testis weight, testis length, testis width, seminiferous tubules diameter and thickness of the epithelium of seminiferous tubules in the experimental group treated with the extract significantly decreased compared with the control group, but in comparison to the diabetic control group, the mean of testis weight, testis length, seminiferous tubules diameter and thickness of the epithelium of seminiferous tubules increased significantly (Table 1; Fig. 2).
Main comparison to the similar studies
300 mg/kg extract of Launaea acenthodes could increase the percentage of sperm progressive motility (slow, fast and without mobility) as compared to diabetic control group. In addition, the mean number and percentage of normal sperm forms in the experimental group increased significantly compared to the experimental group treated with the extract of Launaea acenthods. Aqueous alcoholic extract of the Aunaea acenthodes has powerful hypoglycemic effects and effective compounds in extracts probably increase levels of blood insulin and reduce diabetic blood sugar by stimulating or increasing hyperplasia or hypertrophy in remained B-cells [19]. … [20] Antioxidants protect the sperm cells from damage by free radicals and improve semen parameters [21]. Mean number and percentage of normal forms of head, neck and tail of the sperm in the diabetic samples treated with the extract of Launaea acenthodes significantly increased compared to diabetic control group. Natural antioxidants found in plants lead to spermatogenesis increase and finally increase in the number of sperms by reducing free radicals and reducing oxidative stress in the diabetic rats [22]. Extract of Launaea acenthodes could improve the testis tissue through reducing the number of abnormal sperm, increasing the number of sperms and increasing the thickness of epithelium seminiferous tubules. Diabetes can change the tissue of testis by inducing apoptosis, atrophy of seminiferous tubules, reducing the diameter of the seminiferous tubules and reducing the spermatogenesis cells [11].
Suggestions
More cytological investigations should be done with respect to the mechanism of compounds in the extract of Launaea centhodes in improving sperms parameters and testicular tissues in diabetic samples. Extract of Launaea compounds can be separated and by re-conducting the stages of testing, compounds that have positive effects on reducing diabetes complications in sperms and testicular sperm can be obtained.
Limitations
The effect of different physiological and environmental factors that govern living systems and lack of access to required facilities to determine the exact chemical composition of the extract of Launaea acenthodes were the limitations of this study.
Conclusions
300 mg/kg extract of Launaea acenthodes can reduce the atrophy in seminiferous tubules in diabetic rats and through improving semen parameters and increasing the sperm count reduces the sperm and testicular tissue complications caused by diabetes.
Acknowledgments
The cooperation of Payam Noor University Department of Biology and Agriculture is appreciated.
Conflict of interest
Non-declared
Ethical Permissions
All the processes were conducted based on international guidelines for the care and use of laboratory animals.
Funding Sources
Payam Noor University funded the study.

TABLES and CHARTS

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