Introduction Many studies around the world were performedon herbal plantsfor possible regulatory properties of fertility (1). Commonly herbal plants are used to relieve sexual dysfunction asaphrodisiac, or as agents improving fertility. They enhance sexual performancesuchas via providing of nutritive value(2, 3).E.sativa is an eatable as vegetable orspice, also called rocket, inArabicwhich isnamed “Jarjeer”.
Rocket with many reported propertiesis considered a medical plant such as antimicrobial,renal protective activity, antihyperlipidemic,strong aphrodisiac effect (4, 5). AncientArab, usedE sativa seed in astomach ache and in a therapy for psychosis, while othersdescribed it’s to drawout poison, use in a plaster, such as scorpion poison (6). Althoughdiabetes mellitus experimentally induced by alloxan injection in rats is triedOil of E sativa seeds for prevention and treatment (7), whilein hair loss and burns it is used ointment for treatment (8). Leaves and seeds ofEruca sativa thatpossess a potent antioxidantand prevent oxidative damageby increasingthelevels of antioxidant enzyme(9).Other studies that showed that Rocket have anticanceractivities 10.
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Materials andMethods:- 1-Preparation of alcoholic Eruca sativa leavesextract: Freshvegetableleavesof Eruca sativa were bought from a local market; the leaves were driedin the shaded place for 7-10 days and then powdered using electrical blender. 50gamount ofrocket leaves were placed in a glass percolator with 500 ml of ethanoland were allowed to stand at room temperature for about 72 h. After 3 days themixture was filtered by usingWhatman filter paperandfiltrateextract was concentrated by rotaryevaporator (11).
2-Semensample collectionThis study was carried out in the laboratoriesof the Higher Institute of Infertility Diagnosis and assisted ReproductiveTechnologies at AL-Nahrain University during the period from January to April,2017. Thirty five semen samples were obtained from male withmean age 27, andrange from 20-35 yearsand collected by masturbation in a dry, clean, andsterile disposable Petri-dish after 3-5days period of abstinence in quiteprivate room adjacent to thelaboratory of semen analysis. The container waslabeled with the following information, name, age, abstinence period and timeof sample collection. The samples were placed at 37Cfor 30 minutes to allowliquefaction in an incubator (12, 13).
The liquefied semen wascarefully mixed for few seconds, and then the specimen was examined in detailby macroscopic and microscopic examination within one hour of collection (14). 3-Technique of in vitro human spermactivationCentrifugation swim-up technique was applied inthis study. After liquefaction each semen sample was prepared for SFA, then dividedinto three groups, as control group(G1), low dose ESE (G2)and high dose ESE (G3),these 3 groups of semen samples washed with culture medium and centrifuged for6- 7 minutes at 2400 R.P.
M. The upper layer was discarded. Add 1mL pro-SMARTmedium in sperm pellet slowly in control group (G1). 1mL of the preparedpro-SMART medium supplied with one of two respectively doses ofEruca sativa extraction (50µ g/1mLand100µg/1mL)wereadded to the sperm pellet in treated groups G2 and G3 groups. After incubation for 30 minute at 37C, one drop from the upper layer wasaspirated by Pasteur pipette. Then, examined under light microscope at (400 x)magnification for assessment of sperm parameters.
5- Statisticalanalysis.Means and standard error of mean (mean+SEM)were determined by using statistical descriptive method. MANOVA test wasused to compare between different means. The data were statistically analyzedby SPSS version 24(15).
Results Table(1) shows the percentages sperm motilityfor(G1,G2and G3) post activation whichappeared significantincreased (P?0.05) as compared to pre-activation.Whereastheyshownon significantdifferences (P>0.05) between G2and G3. In general, sperm motility(%)for G1 group was significantly reduced (P<0.05) as compared to G2and G 3groups.
A significantincrease (P?0.05) was reported inpercentages of progressive spermmotility (%) post activation groups in relatingG1, G2and G3as compared with pre-activation group.Also, they showed significant differences (P<0.05) among all groups of postactivation. In general, progressive sperm motility (%) for G3 group was thehighest as compared to G1and G2 groups.
While, G1roup was significantly reduced(P<0.05) as compared to G2 group Nonprogressive spermmotility (%) showed a significant increased (P ? 0.05) in G1,G2and G3 post activation) as compared to pre-activation after usingcentrifugation swim-up activation technique.Whereasnon significantdifferences(P > 0.05) were seen between G1 and G2.On other hand, a significant decreased(P ? 0.05) between G3and G2 groups as compared to G3group.
Thepercentageof immotile sperm revealed asignificantdecreased(p ? 0.05) for all post activation groups as compared to pre-activationgroup.whereasnon significant differences(P > 0.05) were seen between G2 andG3 groups. In contrast significantincrease in G1group as compared to G2, G3groups.DiscussionSperm activation is very essential step inassisted reproductive technologies, that animportant to determining the outcomeon it (16).
inthe present study, the sperm function was improved in human sperm motility andprogressive sperm motility, that related to the culture media and method forsperm preparation can enhancedsperm function in assisted reproductivetechnologies (17). In addition to reported that only the activationmotile sperm will swim-up to the superior area during activation using centrifugationswim-uptechnique (18). Inpresent studyreduction in immotile sperm(%) wasshowedfor all semen samples wereexamined post-activation as compared to pre- activation. However,this resultmay be belong to the preparation method, immotile spermatozoa and semen debris stay in pellet meanwhile, the goodquality spermatozoa were picked up from upper layer and after activation were absent in bad quality spermatozoa(18)and thisresult was agreed with Allaw(19). In the current study, the percentage ofprogressive sperm motility is directly related to treated with ESE, this may bedue to the chemical composition such assaponins,terpenes, flavonoids, steroids,alkaloidsand glycosides were present in the extract were obtained by (20). Moreover, the presenceof some trace elements (Cr,Cu,Fe,Mn and Zn) in the leaves of this plant (21).
Copper(Cu) has been shown to be important forthe activity of an enzyme responsible for removing toxic free radicals. (Cu-Znsuperoxide desmotase) as well as for the activity of phagocytes (22). Furthermore, Jarjeer is act as a good antioxidantssource, such it is contains glucosinolates, carotenoids, phenolic compounds,and degradation products, as isothiocyanates(23).