Effect of Frankincense (Boswellia thurifera)on ...

1 Journal of Health Science,53(4) 365 370 (2007)365 Effect of Frankincense ( boswellia thurifera )onReproductive system in Adult Male RatMohamad Khalid Nusier, ,aHameed Nayef Bataineh,bZiad Mohydeen Bataineh,cand Haytham Mahmoud DaradkadaDepartment of Biochemistry and Molecular Biology,bDepartment of Physiology,cDepartment of Anatomy, Jordan University ofScience and Technology, School of Medicine, Irbid 22110, Jordan, anddDepartment of Biology, Faculty of Science, Jarash University,Jarash 21122, Jordan(Received December 15, 2006; Accepted May 10, 2007)Ingestion of Frankincense [ boswellia thurifera (B. thurifera )] resin at a dose of 250 and 500 mg/kg body weightfor60days by adult male rats was investigated for effects on fertility. Average weights of epididymis, ventralprostate and seminal vesicles increased significantly.

2 Sperm motility and density were also significantly increasedin cauda epididymis and in testes in Frankincense -treated groups. A significant increase of spermatogenesis intestes due to increase in the number of primary, secondary spermatocytes and spermatids in the treatment groupswasattributed to a significant increase in testosterone and Follicle stimulating hormone (FSH). In addition, it alsoincreased the number of implantationsand the number of viable fetuses in female rats impregnated by these males,thereby increasing their histometery of reproductive organs confirmed those words F r ankincense, boswellia thurifera ,fertility, spermatogenesis, male rat, spermatidsINTRODUCTIONF ertility regulation with plant preparations hasbeen reported in ancient literature of indigenoussystems of medicine.

3 A number of plant specieshave been tested for fertility regulation years agoand were subsequently fortified by national and in-ternational 3) Frankincense , also known as olibanum, is ob-tained from a scrubby tree of the genus boswellia ,native to the arid regions of Africa, India and theMiddle East. The gum resin of Frankincense con-tains boswellic acids (BA) and other pentacyclictriterpenes, which have a chemical structure thatclosely resembles that of )It is commonly used in Indian system ofmedicine (Ayurvedic) as an anti-inflammatory, anal-gesic, anti-arthritic and anti-proliferative agent. Inaromatherapy, Frankincense is valued for its effectson the respiratory system . It has been used in steaminhalations, baths and massages for catarrh, bron-chitis and cough.

4 Furthermore, it has been used as Towhom correspondence should be addressed: Department ofBiochemistry and Molecular Biology, Jordan University of Sci-ence and Technology, School of Medicine, Irbid 22110, :+962-2-7201000 (ext. 23696); Fax: +962-2-7095010; E-mail: to ) Frankincense is also used forthe treatment of tumors, ulcers, vomiting, dysen-tery and ,7)In china it is used for leprosy 10)This plant iscurrently used by Jor-danian population as a phrodisiac and fertility pro-moting work was conducted to examine the effectof Frankincense on the reproductive system and fer-tility in adult male AND METHODA nimals andTreatment Thirty adult maleand 60 adult female Sprauge-Dawley rats 3 monthsold weighing approximately 300 gm were bred inthe Animal House Unit at Jordan University of Sci-ence and Technology, School of Medicine, betweenJanuary and May 2005.

5 Rats were maintained undercontrolled temperature of 21 1 Cand12 hr light:12 hr darkness schedule. Food and water were avail-ablead Material Dried resin material [Bos-wellia thurifera (B. thurifera )] from plant was ob-tained from a local supplier. The material was dis-solved in distilled water and administered orally torats using animal feeding intubation needles (Pop-366 Vol. 53 (2007)per and Sons, New York, NY, USA) in concentra-tions of 250 and 500 of LD50in mice was conductedto determine the dose to be given to rats. Gradeddoses of the aqueous extract ofB. thuriferain water was administered intraperitonealy tosix groups of six non fasted male albino mice (25 30 g each). They were housed in transparent plasticcages at 24 C. Mortality was noted after 1 ,12)Experimental Design M a l erats were dividedinto three groups: Control: This group receivedvehicle (distilled water) for 60 days and treatmentgroups 1 and 2, received Frankincense 250 and500 mg/kg body weight for one reproductive cy-cle (60 days), respectively.

6 After 24 hr of the lastdose, animals were weighed and autopsied underlight ether anesthesia. Blood was collected throughcardiac puncture using a dry and clean syringe, forserum Test Fertility was estimated in bothcontrol and treatment groups. Each male rat wasplaced in an individual cage with two virgin un-treated females of the same strain. Animals wereleft together for ten days during which two estrouscycles should have )One week after theremoval of the treated males, females were killed bycervical dislocation under light ether anesthesia andthe number of pregnant females, number of implan-tation sites, number of viable fetuses and number ofresorptions were Motility and Count To d e t erminesperm motility and sperm counts, 100 mg of caudaepididymides was minced in 2 ml of physiologicalsaline.

7 One drop of evenly mixed sample was ap-plied to a Neubauer s counting chamber under cov-erslip. Quantitative motility expressed as percent-age was determined by counting both motile andimmotile spermatozoa per unit area. Cauda epididy-mal and testicular sperm counts were performed byroutine procedure and expressed as )Body and Organ Weights I nitial and finalbody weights of animals were recorded. Repro-ductive tract was trimmed free of fat and each or-gan was weighed separately on electronic reproductive organs of males included testes,epididymides, ventral prostrate, seminal vesicle andvas deferens, were fixed in Bouin s fixative for his-tological Studies T h e Bouin s fixed repro-ductive organs were cut into small pieces and pro-cessed.

8 The paraffin embedding was followed bysection cutting (5 m)and staining (Harris Haema-toxyline and eosin. Sigma Aldrich, St Louis, Mo,USA).Histometry U s i n g C a mera Lucida, hundredcircular appearing seminiferous tubules were tracedat 80 and the diameter of each tubule was mea-sured separately. The measurement was expressedas mean of all the traced tubules. Similarly, Leydigcell nuclei were traced at 800. The epithelial cellheight of cauda epididymides, caput epididymidesand seminal vesicle were also traced at Cell Population Counting S p e r-matogenic , spermato-cytes and spermatids were counted in 5 mthickcross sections of 10 seminiferous tubules in 10 an-imals of each group. All raw counts were trans-formed to true counts by an adaptation of Aber-crombie formula15)from germ cell diameter cell types (such as fibroblast, imma-ture and mature Leydig cells and degenerating cells)were estimated, applying a differential count over200 cells population and statistically verified by thebinomial )Serum Biochemistry To t a l protein, choles-terol, triglycerides, serum aspartate aminotranferase(AST), serum alanine aminotranferase (ALT) weremeasured using commercial kits from Cis BIO In-ternational (AST, ALT from CIS BIO International,Gif Sur Yvette, France)Hormonal Assays Plasma Follicle stimulat-ing hormone (FSH)

9 And testosterone concentrationswere measured by radioimmunoassay using com-mercial kits from Cis BIO Calculation A l l t h evalues ofbody/organ weight, biochemical estimation and his-tometry were expressed as mean value Thetreatment groups were compared with the controlgroup using chi-square test and Student s t )RESULTSE ffect of Frankincense (B. thurifera )on Bodyand Organ WeightTable 1 show s that intragastric administration ofFrankincense (B. thurifera )caused an increase inbody weight, when initial and final body weightswere compared in treatment relative weights of the testes, epi-didymides, seminalvesicle, ventral prostate and vasdeferens were increased 4367 Table and Organ Weights of Male Rat Ingested Frankincense (B.

10 thurifera )ResinTreatmentBody weight (gm)TestesEpididymidesSeminal vesicleVentral prostateVas deferensInitialFinal(mg/100 gm body weight)Control group308 1B. thurifera (250 mg)321 2B. thurifera (500 mg)310 are expressed as mean Ten male rats were included per group. p< , p< significantly different from control group(Student s t test).Table Parameters and Sperm Dynamics of Male Rat Ingested Frankincense (B. thurifera )ResinTreatmentSperm motilitySperm densitySeminiferous tubule(%)(million/ml)diameterTestesCauda ( m)epididymidesControl 1B. thurifera (250 mg) 2B. thurifera (500 mg) cell nuclearEpithelial cell heightdiameter( m)( m)CaudaCaputSeminalepididymidesepididymi desvesiclesControl 1B.