ARTICLE INFO

Article Type

Original Research

Authors

Nejati   V. (1)
Khaneshi   F. (*)






(*) Comparative Histology Department , Veterinary Medicine Faculty, Urmia University, Urmia, Iran
(1) Biology Department, Science Faculty, Urmia University, Urmia, Iran

Correspondence

Address: Biology Department, Science Faculty, Kilometer 11th of Sarv Road, Daneshgah Boulevard, Urmia University, Urmia, Iran. Postal Code: 5756151818
Phone: +984412752740
Fax: +984412753172
f.khaneshi@yahoo.com

Article History

Received:  June  21, 2013
Accepted:  March 1, 2014
ePublished:  April 1, 2014

BRIEF TEXT


Diabetes has negative impacts on males’ reproductive system [4-9]. Some medicinal plants like alluim species (from Liliaceae family) have effects on fertility [10-12]. In the present study, Plantago major (from Plantaginaceae family) has been used, with its therapeutic properties, especially on spermatogenesis increasing in male rats [13-16].

There are some conducted study, concerning impacts of diabetes on reproductive system in males (including testicular tissues) and decrease in production of testosterone [25-27, 30-34].

The aim of this study was to investigate the effect of leaf extract of plantain (Plantago major) on testis morphology, sperm parameters, and serum testosterone levels in the streptozotocin-induced diabetic male rats.

Research type is laboratory experimental.

Research society was the population of 24 male wistar rats with mean weight 200 g to 230 g (Center for Laboratory Animal Care; Orumieh University; Iran). The rats were kept inside a room with 24±2℃ with 14 hours light and 10 hours darkness. Biology Group of Orumieh University conducted the study. Plantain samples were collected from Orumieh Township (Iran) during September 2010; and Systematic Group of Sciences Faculty of Orumieh University identified and confirmed these samples.

To induce type I diabetes, streptozotocin was used. After 7 days from streptozotocin injection, rats with more than 300 mg per dl glucose concentration were identified diabetics. The rats were randomly divided into 3 groups of eight. Physiologic serum, with volume equivalent to the volume of injected extracts to other groups, was injected to the healthy rats of control group (C). The diabetic rats of the second group (DP) were received leaf extract of plantain with 100 mg per kg body weight dose, daily and for 42 days and by the use of insulin syringe and intraperitoneal method in sterile conditions. The diabetic rats of the third group (D) were considered diabetic control. Immediately after treatment period, blood was taken from hearts of the study animals. After the rats were euthanized, lower tail of the epididymis was transferred into a container of physiologic serum and the sperms were isolated. To compute sperm number, the resulting sperm suspension with ratio 1/100 was studied.

Streptozotocin (Sigma, USA) was used. Optical microscope was used to study the sperm suspension [20]. In order to stabilize, testicular tissues were transferred into a container of 10% formalin. After stabilizing the samples and providing tissue cross sections, Hematoxylin-Eosin (H&E) staining and Oil-Red-Oil staining were performed, and the slides were studied by the use of optical microscope. Data were analyzed, using One-way Variation Analysis test and SPSS 16 software. To detect significant mean differences, Tukey Post Hoc tests were used. Epithelium thickness of the seminiferous tubular was computed from the spermatogonias under the seminiferous tubular wall, using scaled lens (in terms of micrometers). Only seminiferous tubules with circular or near-circular cross section were studied [23].

Seminiferous tubules’ diameter, epithelium thickness, and spermiogenesis index in D group was significantly decreased than C and DP groups. There was no statistical significant difference between C and DP groups. Thickness of interstitial tissue between the tubules in diabetic group showed a significant increase than C and DP groups. However, there was no significant statistical difference between C and DP groups. Treatment by the use of leaf extract of plantain in DP group resulted in significant increase in testosterone serum concentration than D group. Nevertheless, there was no significant statistical difference between C and DP groups. The number of Leydig cells in DP group was significantly increased than D group. However, there was no significant difference between C and D groups. Treatment by the use of leaf extract of plantain in DP group resulted in significant increase in sperm number and significant decrease in abnormal sperms percentage than D group. Nevertheless, there was no significant difference between DP and C groups (table 1). In C group, all the seminiferous tubules were apparently fully healthy, and there were all the spermatogenesis cell lines (Fig. 1-A). However, there was decrease in spermatogenesis cell lines in D group (Fig. 1-B). In DP group, seminiferous tubules had normal appearance, and all the spermatogenesis cell lines were regularly fully alongside each other (Fig. 1-C). Leydig cells were normal in C group (Fig. 1-D), as well as in DP group (Fig. 1-F). Nevertheless, these cells were abnormal in D group (Fig. 1-E). Histological examinations with Oil-Red-Oil staining were consistent with the results of measurement of testosterone serum concentration.

In the present study, treatment with extract manifestly increased number of sperms, which is a result consistent with the reported effects of onion on diabetic rats [12]. Morphologic findings of the present study are consistent with reported effects of pomegranate juice on testicular tissues of the diabetic rats, enhancing spermatogenesis procedure and increasing seminiferous tubule diameter, while decreasing diabetes’ side effects [33].

It is suggested that more cytological investigations to be conducted, concerning function mechanism of the extract compounds on reproductive system.

Non-declared

With its direct impact on the testicular tissue, plantain makes diabetes’ side effects to be in minimum level; and improving testosterone serum concentration and increasing spermatogenesis, it increases fertility in diabetic persons.

Researchers feel grateful to Research and Education Deputy of Orumieh University.

Non-declared

Non-declared

Research and Education Deputy of Orumieh University funded the research.

TABLES and CHARTS

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