Ir aqi J.Pha rm.Sc i., Vol.15 (2 ) ,2006 Sex Hormones Chan ges
23
The Chages in Sex Hormones in Female Working in Battries
Manufacturing Plant
Mohammed A. Taher *1 , Salim A.Hammadi** , Ali A.Ali**
*De pa rtme nt Clinica l Labo ra tory Sc ie nces, Co lle ge O f ph armacy ,univ ersity of Bag hdad, Baghd ad -Ira q.
** Depa rtm ent Clin ical Pha rm acy , C olle ge Of pharm acy ,un iversity of Baghdad, Baghdad -Iraq
Abstract
Lea d has toxic e ffects on reproduction of both male and fema le . It can c aus e dec re ase d sex
drive , infertility a nd abnormal menstrual cycle in women. This study wa s des igned to
eva luate the effec t of e xposure to lea d in batte ries female workers on s ex hormones leve l in
the s erum.Thirty nine (39) fe male worke rs (voluntee rs ) in Iraqi Batte ries Manufa cturing
Plants, Al-Waziriya / Ba ghdad we re pa rticipated in this study. The y a re clas sified into 3
groups, first group inc luded fourteen (14) fema le tha t have be en employed for 1-7 yea rs ,
se cond group include d thirte en (13) fema le that have bee n employe d for 8-14 yea rs , third
group included twe lve (12) fema le have be en employed for15-22 years and and fourtee n
fe males were include d as the control . Blood lead leve l, serum FSH, LH, prolac tin and total
te stosterone we re me asured a nd compared for a ll subje cts.The results indica te d that me an of
blood lead leve ls (BLL), te stos te rone levels were highly s ignificant in a ll worke r groups
compared to the control (p<0.005).Prolac tin le vels in group I a nd FSH in group III were
signific antly highe r than that in c ontrol ( P< 0.005) a nd (P<0.05) res pectively. LH leve ls in
groups II and III were signific antly higher than that in control (P<0.05, P<0.005 res pec tive ly).
High incidence of hirsutism (48%) and mis ca rria ges (50%) were obs erved in worker groups
compared to c ontrol (11%). The res ults indica te d that there are hormonal changes in fe male
workers expos ed to lea d as sociated with increa sed inc ide nce of hirs utis m a nd misca rria ges
compared to non expos ed fe males .
K ey words: Lead , Sex hormones hype randroge nemia
لخالصة ا
عات قسمت العامالت ن في هذه الدراسة كمتطو ريات في بغداد شارك مل صناعة البطا ن امرأة عاملة في مع عة وثالثو تس
ص رصا رة التعرض لل مادا على فت ميع اعت جا .الى ثالث م
ملة وفترة الخدمة تتراوح بين ) ١٤(المجموعة األولى تشمل - ۱ ر سنوات) ۷ـ١(عا .سنة) ٥۰ـ٢٤(وبعم
عة الثانية تشمل ۲ مة تتراوح بين ) ۱۳(ـ المجمو خد رة ال سنة ) ٥٥ـ۳۰(سنة وبعمر ) ۱٤ـ۸(عاملة وفت
عة الثتلثة تشمل ۳ راوح بين ) ۱۲(ـ المجمو .سنة) ٥۲ـ۳۲(سنة وبعمر ) ۲۲ـ۱٥(عاملة وفترة الخدمة تت
رابعة- ٤ ۱±۲۹( امرأة بمتوسط عمر) ۱٤(مجموعة السيطرة وتشمل : المجموعة ال .نةس) ۷.
ها بمجموعة ر في مستوى الهرمونات الجنسية عند النساء العامالت عند مقارنت غي ن هنالك ت وجد في هذه الدراسة ا
رة طويلة .السيطرة ص لفت رصا عامالت اللواتي تعرضن لل ستوى الهورمون اللوتيني في المصل يزداد عند ال كذلك ان م وجد
عن ريب في المصل يزداد فقط رة طويلةاما هورمون محفز الج ض لفت ن .د النعر والكتين (اما هورمو عد ) البر فيزداد فقط ب
ض لفترة قصيرة للرصاص حبا مع زيادة في .التعر زداد عند النساء العامالت متصا كذلك هورمون التيستوستيرون الكلي ي
مقارنة بمجموعة السيطرة .نسبة الشعرانية واالسقاطات عند العامالت
1c or res po nding au th or e mail Moha mmed – Ta her 4 3 @ya hoo .c om
Rec eived 21 -11-200 5
Acc ep te d 7-10-2006
Ir aqi J.Pha rm.Sc i., Vol.15 (2 ) ,2006 Sex Hormones Chan ges
24
In troduction
Some sc ie ntis ts cautioned the Romans of the
da nge r of inhaled fume s from lea d sme lting (1)
With the industria l re volution, le ad po is oning
be came a common oc cupationa l problem .The
re productive effe cts of lea d p oisoning were
also noted by the turn o f the ce ntury and many
artic le s d es crib e the high rate of s tillb irths,
infe rtility and a bortio ns among wo men in the
pottery ind us try, o r who we re married to
pottery worke rs (2).
The mos t imp orta nt route of
ab sorption in o ccupa tional s etting is thro ugh
inha la tion o f lead d us t and fumes . In add ition,
worke rs ma y ea t, drink or s moke in lea d-dus t-
co ntamina te d areas re sulting in so me inges tio n
as we ll. S to ra ge ba ttery ma nufac ture inv olve s
co nside ra ble exp os ure to lead o xide dust, in
ad dition to fume s from we ld ing o f battery
co nne ctor (3) .
Lea d c an ca use de crea sed se x d rive and
infe rtility in wo men. In a dditio n, it c an ca us e
ab normal me nstrua l c ycles (dys menorrhea,
menorrhagia and ame no rrhe a), prema ture birth,
sp ontaneo us misc arriages , and stillbirths . The
incide nce of p olyme norrhea , prolonged and
ab normal mens truatio ns , hypermenorrhea wa s
significantly higher in the lea d expos ed group
(female workers of le ad ba tte ry p la nts) than in
co ntro ls (4). This s tudy was des igne d to
ev aluate the hormonal changes (LH, FSH,
prolactin and tes to sterone) in w omen whom
expos ed to lead in b atte ries ma nufa cturing
plants through inhala tion and d irec t contact
with a ctiv e co nstituent of batterie s.
Subjects and Methods
This s tudy was carrie d out on female
worke rs emp lo ye d in Iraq i b atterie s
manufac turing p la nts, Babylo n 1 and 2 in Al-
Waziriya / Baghda d for the p eriod o f thre e
months from J anuary to April of 2 005 .
Thirty nine (39) fema le workers were
pa rtic ip ated in this study , they work 6 ho urs
pe r d ay eve ry othe r d ay and had b ee n
emp lo yed for at le ast 1 ye ar in the p la nt .
The subjec ts we re c la ss ified into 3 gro up s
ac cording to the dura tion o f expos ure to lea d
(e mployment in the plant) a s follo w:
Group І ,inc lude s fourtee n (1 4) female that
ha ve be en emp lo ye d for1-7 ye ars with ra nge
age (2 4-5 0) yea rs (29.2± 7 .2 ) ye ars.
group П, inc ludes thirte en (1 3) female that
ha ve be en e mploye d for 8-14 yea rs with ra nge
age (3 0-5 5) yea rs (37.9± 7 .9 ) ye ars.
gro up Ш includ es twelve (1 2) female that
ha ve bee n e mployed fo r 15-22 yea rs with
ra nge (32 -52 ) years (4 1.1± 6) yea rs .
Fourtee n he althy wome n, not e xpo se d
previously to lea d with a ge range (2 4-5 0 )
ye ars (2 9±7 .1 ) were utilized as c ontrol.
Individual q ue stio ne r protoco l was followe d
fo r all wome n conce rned with gyne cologic al
and o bste tric al his tory including marrie d or
not , age o f marriage , number o f children ,
type of de live ry (no rma l o r ca es are an s ec tion) ,
numb er of mis ca rriage s, growth o f the ir
infa nts , if they work d uring p re gna nc y ,
re gular or irregular me nstrua l cyc les , if they
are with ame norrhea , d ysmeno rrhe a or
meno rrha gia . Ap propriate da y fo r ea ch fema le
(b etween 2 nd a nd 5th d ay of mens trua l cycle)
was se le cted to co ns id er follicle phas e for
FSH and LH as sa y , and female were ad vised
to fas t 1 2 hr. be fo re s amp ling , fo r ap pro pria te
ana lysis of prolactin . Blood s ample s (1 4 ml)
were drawn from ea ch p atie nt a nd c ontrol by
vein puncture left to c lo t a nd se rum was
sep arated b y ce ntrifuga tion.
Blo od lea d le vels we re meas ured using the
slotte d q ua rtz tube metho d.(5).LH ,FSH and
prola ctin le ve ls in se rum we re a na lyze d using
ra dioimmunoas sa y method s(6,7,8) while
te stos te ro ne was as se ss ed ac cording to the
metho d o f Abraha m e t al (9). All thes e kits
sup plie d by Immunotec h , A Beckman c oulter
comp any (France).
Inde pendent t-te st wa s us ed to examine the
difference in the mea n of control and wo rkers,
also the d ifferenc es amo ng worke r gro ups
thems elves .P-v alue s < 0.05 we re considered as
signific antly diffe re nt. Pe ars on c orre la tion (r)
was pe rformed to find re la tio nship be tween
exp os ure time and tes toste rone leve ls .
Results
Table (1) s hows the a ge s and o cc upa tional
(e xpo sure) p erio ds of fe male worker. The
numb er of ma rried were 3 ,6 ,7 in I, II, III
group s re spe ctively .The me an a ge of c ontrol
group wa s 29.1 ±7 .1 yea rs a nd the numbe r of
married wa s 5 .
Table (2)shows me an b lo od lead le vels in all
group s of wo rkers are signific antly (p<0 .0 05)
highe r tha n that of the c ontrol gro up. Gro ups II
and III worke rs hav e significantly higher blood
le ad le vels than that of group I worke rs
(p <0.05 ) . Se rum to ta l tes to sterone le vels in a ll
thre e worke r group s were signific antly higher
in c ompa riso n with that of the co ntro l group
(P<0.005 ). Total te stos te ro ne lev el for gro up I
workers wa s the highes t one. Howev er gro up I
workers ha ve a s erum to ta l tes to sterone lev el
that was significa ntly higher than that of
group s II and III workers (P<0.05).Mean
prola ctin le ve l in group I worke rs sho ws highly
signific ant inc re as e c omp ared to tha t of the
control group (P<0.005 ).While mean p ro la ctin
le vels in gro ups II and III were non-
signific antly elev ated co mpa re d with the
control group (P>0 .0 5).Group I worke rs have
prola ctin le ve ls
Ir aqi J.Pha rm.Sc i., Vol.15 (2 ) ,2006 Sex Hormones Chan ges
25
significantly higher than that of groups II and
III of wo rkers (P<0.05).Mean F SH lev el of
group III workers was s ignifica ntly e le va te d
(p <0.05 ) in comparison with that of the control
group. While me an FSH le vels of wo rkers
group I and II we re non-significa ntly e le va te d
(P >0.05 ) comp ared to co ntro l group . .F SH
le ve ls s how no n significant diffe re nce a mong
worke r group s thems elve s .Me an LH leve l in
group II wo rkers was e le va te d signific antly in
co mpared to that of the c ontrol group. LH
le ve l in group III wo rkers s ho ws a highly
signific ant increa se compared with c ontrol
group (P<0 .0 05).Furthermore mea n LH lev el
in group I workers wa s non-s ignifica ntly
eleva te d (P>0 .0 5) c omp ared to tha t of the
control group .LH lev els show non s ignifica nt
(P>0.05) d iffe re nc e a mong worke r gro ups
thems elves . There are non s ignifica nt
differences a mong LH/F SH ra tio in a ll three
group s of wo rkers and contro l group
(P>0.05).Als o LH/FS H ratio show non
signific ant (P >0.05 ) diffe re nce a mong w orker
group s.
Table (1)De mo graphic data of wo rke rs and c ontro l wo me n.
Table (2)Se rum le ve ls o f le ad, tes toste rone , prolac tin, fo llicle stimulating hor mone (FS H)
le utiniz ing hor mone (LH) in fe ma le s working in batte ry indus trie s.
Parame te rs Control Group I Gro up II Group III
Le ad(µg/d l) 13 .4±4 .5 2 1.4±6.7** a 31 .6±7 ** b 29.7±7.3** b
Testos te ro ne
(ng/ ml)
0 .156±0.5 0.45 8±0 .15**a 0.35 6±0 .13**b 0.28±0.1** b
Prolac tin
(ng/ ml)
8.5± 4.1 19.7 ±11 .9 ** a 1 0.7±7.2NS b 11.7±8.1 NS b
F SH (mIµ/ ml) 6.9± 2.7 13.3 ±14 .8 NS a 10.9±8.4 NS a 14.4±13.6* a
LH ( mIµ/ml) 8.8± 3.3 1 1.1±3.7 NS a 14.7±10.4* a 17.3±9.1** a
LH/FS H 1.2±1.64 0.83 ±0.45 NS a 1.3±1 NS a 1.2±0 .85 NS a
Value s are expre ssed as mea n±SD
* p<0 .0 5 significant diffe re nce fro m c ontrol g roup
**p<0.005 high ly s ig nifica nt diffe re nce from co ntrol gro up
Values with diffe re nt le tters (a,b) a re s ig nifiga ntly d iffe re nt (P<0.05).
Figure (1) s ho ws the co rrelation be twe en time
of le ad expos ure and se rum te stos te ro ne lev els
in fe male workers.A ne gative c orrela tion wa s
fo und b etween time of lea d e xp osure and
se rum te stoste ro ne le ve ls (r=-0.43), (P<o.o5).
In table (3) all wo rkers groups hav e
perce ntage o f hirs utism (17 .9 %) and
mis ca rria ge s (5 0%) higher than that in c ontrol
group . In ta ble (3) all worke rs gro ups have
perce ntage o f hirs utism (17 .9 %) and
mis ca rria ge s (5 0%) higher than that in c ontrol
group .
Table (3)Distribution o f fe male worke rs with hirs utis m a nd miscarrige s
Groups Age
(me a n ±SD)
(yea rs )
Oc cupatio n
pe riod (me an
±SD)( ye ars)
Numbe r of
marrie d
Numbe r of
unmarrie d
Total
I 2 9.2±7 .2 3.6±1.9 3 11 14
II 3 7.9±7 .9 1 1.1±1.4 6 7 13
III 4 1±6 1 8.2±2.8 7 5 12
Control 2 9.1±7 .1 - 9 5 14
Va riable s Gro up I
No. (%)
Group II
No. ( %)
Gro up III
No. ( %)
Total Co ntrol
Hirs utism 2 (14) 2(15 ) 3(25) 7(17 .9 ) 0 (0 )
Miscarriages * 1 (33) 3(50 ) 4(57) 8 (50) 1(11)
Ir aqi J.Pha rm.Sc i., Vol.15 (2 ) ,2006 Sex Hormones Changes
26
Time of exposure (years).
S
e
ru
m
t
e
st
o
st
e
ro
n
e
(
n
g
/lm
l)
.
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
-2 2 6 10 14 18 22 26
Discuss ion
The re sults indica te d that mea n of blo od lea d
le ve ls (BLL) were s ignifica ntly highe r in a ll
worke r group s comp are to the c ontrol group
(p <0.00 5),this may be d ue to lo ng term
expos ure to le ad oxid e which use d in b atterie s
manufac turing p la nt. The wo rkers practice in
impro pe r environment, they we re not we aring
fa ce ma sk, clov es with b ad ve ntila tion ins id e
plant ro oms , in add ition to that wo rkers were
ea ting ins id e the manufac turing rooms, whic h
mea ns the y d id not follo w the co nd itions of
oc cup atio na l sa fe ty. The wo rkers in group I
ha d blood lea d le vels s ignifica ntly lo wer tha n
that in gro ups II a nd III ( P< 0.05 ),this may b e
due to longer e xpo sure time in groups II and
III tha n that fo r group I
It has b een re porte d that pa tients with bloo d
le ad leve ls les s than 4 5µg/d l do no t re quire
chelatio n therapy (10). The me an bloo d lea d
le ve ls in mo st fe male worke rs a re le ss tha n that
re ported in prev io us stud ie s (11,12 ), b eca us e
fe males in prese nt s tudy are working ev ery
othe r da y in the plant sinc e 1.5 ye ars ,b eca us e
of s ho rtage of th raw ma te rials
The me an se rum tes to sterone le vels in a ll
worke r group s were signific antly higher tha n
that of co ntro l gro up (P<0.005 )(table 2 ).
Figure (1) d emo nstrated a negativ e correlatio n
be twe en time o f expos ure to lea d and s erum
te stos terone lev els in worker group s .The d ata
showed a n inve rs e re la tionship be tw een time
of le ad expo sure a nd s erum te stos tero ne lev els,
this may need further inves tigatio ns to explain
the correlatio n. The sa me res ults hav e bee n
re ported by Sokol-Rz (13), where the rats
trea ted with v ario us do se s of lea d ace ta te fo r
more tha n 1 wee k e xhibited a s ignifica nt
increas e in Gona dotropine relea sing hormone
(GnRH) mRNA, b ut with a ttenua tion of the
increas e at highe r co nce ntratio ns of le ad with
increas ed d uratio n of exp os ure.The y co ncluded
that the s igna ls within a nd betwe en the
hyp otha la mus a nd pituita ry gla nd a ppe ar to be
disrupted b y long-term, le ad expos ure.
LH s timulate s the ca cells o f ova ry to
pro duce tes to sterone in female (14).Ho weve r, in
this s tudy, it ha s bee n fo und tha t as LH lev el
increas es , tes to sterone le vel d ecrea se s (tab le
2), indica ting tha t elev atio n of tes to sterone
le vel in fema le wo rkers may b e extra ov aria n.
Howev er most of stud ie s co nce rning lead we re
app lied o n a nimal mode ls s o it is d iffic ult to
comp are p re se nt findings with their res ults.
Gro up I hav e a high mean p rolac tin leve ls
which are signific antly higher than tha t of
control group ( P< 0.00 5); while mean
pro la ctin le vels in group II a nd gro up III we re
not s ignific antly different from co ntro l group
( P>0 .0 5) (table 2) .Prolac tin relea se fro m the
pituitary is und er tonic inhibito ry c ontrol fro m
hyp otha la mus -de rive d dop amine or p ro la ctin
inhibito ry fac to r (PIF). Thyrotro pin-re le asing
hormone (TRH) in turn is s timula to ry to
pro la ctin relea se .Estro gen c an dire ctly
sensitize the pituita ry to re le as e p ro la ctin (15).
The elev atio n of prolac tin le ve ls in w orker
group s could b e attribute d to tha t le ad a ffec ts
dop amine rgic co ntro l of prolac tin sec re tion
from pituitary gland, acc ording to p re vious
study on rats (16).
The results d emonstra ted that only group
III workers hav e me an serum FSH lev el
signific antly highe r than that o f the c ontrol
group (P< 0 .0 5) (ta ble 2 ).The e le vation in
serum FSH als o rep orte d b y Ng et al. (17)
where significantly higher s erum FSH and
serum LH lev els were o bse rv ed in lead -b atte ry
male workers , d uring le ss tha n 10 yea rs
exp osure p erio d , whe re as tho se expo sed for
10 ye ars o r more showed normal s erum LH
and s erum FS H conce ntra tions.Howe ve r,
Viv oli G et al(18) re ported nega tive
re la tionships betwee n blood lea d le vel and
LH and F SH in ma le s with lea d le vels higher
than 9µg/dl.
Group I worke rs a re pres ente d with se rum
LH not significantly different from that in
control group (P> 0.05 ) while group II a nd III
hav e mea n serum LH s ignifica ntly highe r than
that in co ntro l group (P <0.05 ,P<0.005
re spe ctiv ely) (tab le 2 ) . This elev ation in se rum
LH le vels are in agree ment with that rep orte d
by Rod amila ns e t al.(19) who rep orte d tha t in
le ad-sme lte r workers, s erum LH lev els a re
signific antly raise d, a s co mpa re d with c ontrols.
Figure ( 1 ) : Co rre latio n be twe en time
of le ad ex pos ure and se rum
testos tero ne .
Ir aqi J.Pha rm.Sc i., Vol.15 (2 ) ,2006 Sex Hormones Changes
27
Furthe rmo re Yen (20) sugge sted tha t ba sal LH and F SH le ve ls gra dua lly ris e after the age of
ye ars 35 and c ontinue to climb until se veral
ye ars after onse t of menopa use due to dec rease
ne gative fee dba ck c ontrols on p ro ductio n.
Ronis et a l. (21) s tated that, in fema le ra ts
exposed to le ad prep uberta lly , dela y vaginal
opening with mo re se vere reproduc tive
disrup tion, acco mpa nied by suppression of
circ ulating es tradiol .Effe cts on c irc ulating se x
steroids were accomp anie d by va riab le e ffec ts
on circulating LH le ve ls , p ituitary LH, and
pituita ry LH b eta-mRNA. Inc re ase in
hypotha la mic le vels of Gn RHmRNA, and a n
increase in pituita ry leve ls of LH mRNA and
pituita ry s to res of LH in lead dosed a nimals.
This inc re ase due to lead disrup ts the
re productive axis by interfe ring with fe edb ac k
mec hanisms a t the hypotha la mic and p ituitary
levels (22).
In this s tudy the re sults indica ted that lead
expos ure has no e ffec t on LH/FS H ratio in
fe male workers .Elevated basa l LH with a n
LH/FSH ratio >2 and some increase o f ovaria n
androge n in a n esse ntia lly no n ovulatory adult
wome n is p resumptiv e evidence of polycystic
ovary s yndro me. A high se rum F SH to LH
ra tio (1 .9 to 3.8) has been obse rved in post
menopa us al wo men. Hyp ogonadism is usually
associated with increased bo th F SH a nd LH
levels ; while dec reased FS H and LH ma y
occur in p ituitary or hyp otha la mic failure (23).
Tab le (3) showed high incidence o f hirs utism
(4 8%) in wo rking women in batteries plant
than in co ntro l group (0%). Als o pe rcenta ge of
hirs utism in female worke r groups increa sed
with inc re asing expos ure time to lea d. S ome of
worke rs had hirsutis m, the ir s erum tes to sterone
levels we re high, and this typ e o f hirs utism
se ems to be a nd ro gen induced hirs utism
(hyp era nd ro genism). In this stud y in s pite of
decreased se rum tes to sterone levels with
prolonged e xp osure ti me (fig.1), pe rcenta ge of
hirs utism inc re ase d as time of exp os ure
increase d ( table 3), so it ca n be conc luded
that hirsutis m may d epe nd on many fa ctors
othe r than and ro ge n (te stos terone ) le ve ls in
fe male exp osed to lead .Whe n hirs utis m is
associated with o besity a nd menstrual
ab normalities , the sourc e of a nd ro gen e xc ess is
ofte n ov aria n, typica lly p olycystic ovary
syndrome . Whe n it is a ss ocia ted with average
weight and no rma l menses, the source is ofte n
ad re nal and ra re ly (in <5 % o f cases)
pituita ry(24). Tab le (3) showe d increa sed
incide nce o f mis carria ges in fe male worker
groups (50%) c omp ared to control group
(1 1%). Le ad has toxic action o n the
trop hob lastic ep ithe lium and to nic contra ctio n
of the uterus . It the refore results either in
abortion o r a de ad fetus (25).
In this s tudy whe n lead expos ure time
increased , perce ntage of misc arriages also
increased (tab le 3).Much of the previous
litera tures focuse d o n an increase d incide nce
of sponta neo us abortio n a nd stillb irth
assoc ia ted with le ad exposure in the workp lace
(25 , 26).Other s tudies hav e examine d the is sue of
lead's invo lv eme nt in s pontaneous abortio n,
stillbirth, preterm d eliv ery, and low b irth
weight. Women in the s tudies in Bos to n (27),
Clevela nd (28), Cinc inna ti (29), and P ort Pirie (30)
had average blood lead co ncentratio ns during
pre gnancy of 5 -10 µg/dl; almost all ha d blood
conce ntra tion less than 25 µg/dl. In the
Cincinnati stud y, ges ta tional a ge was re duced
abo ut 0.6 wee ks fo r eac h natural log unit
increase in b lood lead concentra tions.
However, the Cincinna ti and Port Pirie s tudies
fo und a le ad related decrease in duration of
pre gnancy, and Cinc inna ti and Boston s tudies
re ported a lead-related decrease in b irth
weight .In conclus io n there a re e vide nt
cha nges in s ex hormones due to expos ure of
fe male wo rkers to lead in Batte ry
manu fa cturing plant.
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