Introduction: Previous surveies have shown that morphia ingestion during gestation may detain embryo development or cause unnatural nervous system map. The present survey focused on the consequence of maternal morphia ingestion on development of maternal and foetal part of placent and blood corticosteron concentration in habit-forming pregnant female parent.
Material and methods: In this research 24 female Wister rats, 170-200g weight, were used. The experimental groups after gestation received an unwritten dosage of 0/05mg/ml of morphia by tap H2O while the control group received tap H2O. On 10th and 14th twenty-four hours of gestation, Wister rats were anesthetized by trichloromethane, and placenta with uterus removed surgically, 1ml blood was collected from each pregnant female parent from Retro-orbital fistula, the concentration of blood corticosteron was determined by corticosteron Elisa kit after centrifugation. The fixed embryos underwent tissue processing, segmenting and staining with hematoxylin and eosin ( H & A ; E ) . Placenta was studied microscopically harmonizing to the thickness of beds, country of blood cisterns, and the figure of cells in maternal and foetal part of placenta.
Consequences: Comparing the plasma corticosteron concentration of the intervention and the control groups, non merely a terrible addition in the intervention group was detected, but besides the thickness of maternal and embryologic parts of the placenta at twenty-four hours 10th and 14th of gestation ( pregnant ) was different significantly ( pa‰¤0.05 ) . Furthermore, an addition in figure of cells in maternal and embryologic part of placenta and a lessening in blood cistern country were demonstrated in both the experimental and the control groups.
Decisions: The effects of morphia, including an addition in blood concentration of corticosteron, in dependent pregnant female parents was seen. development of placenta in the experimental group with was delayed
Cardinal words: Placenta foetal part, Placenta maternal part, Blood cisterns, Morphine, Rat, corticosteron
Dependence on habit-forming drugs spread all over the universe and the side effects of dependence non merely affect the 1 who is on drug but besides others are indirectly affected particularly placenta and foetus ( 1 ) .
It is necessary to analyze the map of drugs in animate being tests particularly in placenta.
Many behavioural jobs in addicted female parents ‘ babies indicated the effects of opioides in embryo. ( 1,2 ) .The bulk of surveies focused on the embryo, whereas they neglected to analyze the placenta as an of import organ.
Disruptive effects of ingestion of opioides in human samples and research lab animate beings were good conducted. The research showed that ingestion of opiate stuffs by pregnant female parents cause hold in embryologic development and malfunctions, such as spinabifida ( 1,3 ) . In conformity to old surveies, high blood coicosteron concentration of pregnant female parent attenuate placenta and embryo. The capacity of placenta for supplanting and let go ofing nutrient stuffs depends on the placentas, form, size and reassigning factors. As morphia is little and beseeching molecule can easy go through through blood barrier and placenta and so go effectual on embryologic cells. ( 4,5,6, ) .
As placenta in mammals is the most of import portion to exchange stuffs between embryologic and maternal blood, the size of the placenta is straight related to nutrient stuff transporting ( simple an active conveyance ) ( 2,6,4 ) .The morphine effects were presented with Mio, Kappa and Delta opioides receptors and triping of these receptors caused several alterations, including lessening in the CAMP, an addition in end product of -K+ ion and a lessening in input of ca-ion. ( 8,7 ) On the other manus, the ca-ion has of import function in secernment of estrogen and Lipo-Lutin endocrine from placenta, stability and embryologic development. ( 9,10 ) By advancement of gestation, placenta can move as a secretory organ that secrets Lipo-Lutin, estrogen and the similar that are needed for embryologic development and considered as an option for ovary secernment endocrines. ( 11,6 ) Therefore, morphia can move as an interfering that causes disfunction in placental secernment operating and hold in placental development. ( 10,5 ) Several experiments have shown that morphine disposal cause the lessening of placenta weight in coneies, ( 12,13 ) The presence of opioides receptors on the placental villi cells can impact placental map.On the other manus, because the placenta is a protective barrier, it can forestall the input or end product of some stuffs. Placental barrier as a protective mechanism is frequently considered against infective factors. ( 14,2 ) Disorders in the development of placenta cause placental disablement in interchanging, hormone and protective Acts of the Apostless in embryo ( 6, ) . Corticosteron endocrine stimulate morphia map ( 6,9 ) The importance of blood mother`s corticosteron concentration in placenta development and the effects of morphia on hold of placental development are the major grounds for the present research that consider the unwritten morphia disposals consequence on the placenta of habit-forming female parents on 10th and 14th yearss of gestation.
Materials and methods:
Animals, female Wister rats were used 170-200gr weight. The animate beings were housed 2/cage to a temperature of ( 24 A±10a„? ) and lighted controlled environment with a 12-h light/dark rhythm and provided with nutrient and H2O.
In this survey, prepared morphia sulfate from Iran tamad Co, was used orally. Twenty-four female Wistar rats were divided into four equal groups dwelling of six animate beings each
and the 24 female rats were mated into 2 groups with one grownup male rat. After doing certain about gestation ( with the observation of vaginal stopper and being of sperm in vagina ) , they were separated from male rats the following forenoon and kept in the same coed-groups. Thereafter ( 0 twenty-four hours of gestation ) , experimental group received a day-to-day dosage of 0.05 mg/ml ( 15 ) ( 5 milligram morphia per 1000 milliliter drinkable H2O from metropolis grapevine for 12 rats ) The sum of unwritten morphia were 14ml /100gr rat organic structure weight.also, 1 milliliter blood were collected from retro-orbital fistula at 13th yearss of gestation. Blood samples were centrifuged. And plasma was extracted and maintain in -20`C. eventually plasma cortcosteron was assessed by Rat Corticosterone ELISA kit ; EIA-4164 ; DRG Instruments GmbH, Germany ) in both experimental groups.
In 10th and 14th yearss of gestation, they were anesthetized with trichloromethane and placenta were removed from female mice and transferred to 10 % formalin solution for 10 yearss, so, placenta were put on in tissue processing system and were ready for modeling, so blocks were sectioned with microtome and sagital subdivisions hundred of 5I?m thickness serially ( 15,16 ) . These pieces were put on slides and stained with hematoxylin and eosin methods ( H & A ; D ) . After staining, slides were studied microscopically.
Datas were reported as average A± S.E.M. Differences between all groups were calculated by a one-way analysis of discrepancy ( ANOVA ) and post-hoc Duncan trial by utilizing the SPSS/PC computing machine plan ( version 9.1 ) . Statistical significance between the two measurings was determined by the two-tailed odd sample T-Test Result were considered Statistically important when P & lt ; 0.05.
The thickness of part placenta, blood cisterns surface, and figure of cells in the experimental and control groups was measured with motic package. The system used included a microscope connected to a computing machine and a proctor by package could take exposures from slides. Subsequently, the figure of cells on each bed was counted indiscriminately and compared with that of the experimental groups.
Our consequences showed the unwritten morphia ingestion at pregnant mice increased the placenta concentration of corticosteron at twenty-four hours 13 of gestation in comparing with control group. In add-on morphia disposal in pregnant rats placenta showed, thickness of maternal portion has increased in 10th and 14th yearss placentae ( Tabel 1 ) in contrast, important decrease in thickness of placenta embryologic portion was shown ( Tabel 1 ) . besides, increase in cells figure of placenta maternal and embryologic portion in experimental groups was assessed ( Tabel 1 ) thereby decrement in blood pools surface of placenta maternal and embryologic portion in 10th twenty-four hours experimental groupswas shown in this survey ( Tabel 1 ) .
Day Thickness ( Aµm ) Number ( count/ unit ) Area ( Aµ A? ) milliliter
Pregnancy Maternal Fetal Maternal cell Fetal cell Maternal blank Fetal blank Concentration
Control placenta 10th 873.85 A±114 677.6 A±36 13 A±0.1 5 A±0.1 45307 A± 0.1 27046. A±0.1
Experiment placenta 10t 1064.55 A±197* 205.2 A±33* 13 A±0.1** 7 A± 0.1* 20300 A± 1210** 19525 A± 491**
Control placenta 14th 574.92 A±26 1394.67 A±0 17 A±1 17 A± 0.5 21979.3 A±317 7970.54 A±164
Experiment placenta 14th 2351 A±173*** 533.1 A±0*** 24 A±5** 24 A±2* 21935.5 A±242 8383.1 A±118
Control Plasma 13th 650 A± 45
Experiment Plasma13th 1230 A±52**
Consequence were considered Statistically important when P & lt ; 0.05. ( Average A± SEM )
( Placenta Portions Thickness in Tabel 1 ) : Consequence of disposal of unwritten morphia on the10, 14th yearss of placenta development and comparing of control and experimental group, in the thickness of maternal and foetal part of pregnant Wistar rat placenta. The important addition in the thickness of maternal part ( 10,14th yearss ) and important lessening in 10,14th yearss foetal part of placenta in experimental ( E ) group to command ( C ) group. ( *P & lt ; 0.05, **P & lt ; 0.01, **P & lt ; 0.01, ***P & lt ; 0.001 )
( Cell Number in Tabel 1 ) : the consequence of unwritten morphia ingestion on development of 10 to 14-day old placenta and comparing of them in the figure of cells in maternal and foetal part placenta of gestation Wistar rat. The important addition in the figure of 10,14th yearss placental maternal part and figure of 10th twenty-four hours foetal part cells in experimental ( E ) group to command ( C ) group. ( **P & lt ; 0.01, *P & lt ; 0.05, **P & lt ; 0.01 )
( lacuna Area in Tabel 1 ) : Consequence of disposal of unwritten morphia on the10,14th yearss of placental development and comparing of them in control with experimental group, in the blood cisterns ( blank ) surface of maternal and foetal part of Wistar rat placenta. The important lessening in the blood cisterns surface of 10th twenty-four hours foetal part of placenta and the blood cisterns surface in 10th twenty-four hours maternal part of placenta in experimental ( E ) group to command ( C ) group. ( **P & lt ; 0.01, **P & lt ; 0.01 )
( Plasma concentration in Tabel 1 ) : Plasma corticosterone concentration in rats which received unwritten morphia on the thirteenth twenty-four hours of gestation ; The experimental ( E ) and Control ( C ) groups. As it is clear from the figure, a important addition in plasma corticosterone concentration was observed in the experimental group. ( **P & lt ; 0.01 ) different from control group.
Morphologic observations on 10-day old placenta:
Microscopic observation indicated that on the 10-day old placenta, experimental group had small thickness on embryologic portion, more thickness on maternal portion, more cell figure at maternal and foetal portion, and less country blood cisterns ( blank ) in comparing to command group ( A1, A2, A3, A4, B1, B2, B3, B4, ) .
Fig B1: Morphologic alterations of placenta portion thickness decrease and maternal portion thickness incrementin experimental ( A1 ) groups in the 10-day old placenta by A-40 ( two arrowheads ) .
Fig B2: Morphologic alterations on 10-day old placenta in experimental group, bespeaking increase in cells figure of placenta maternal and embryologic portion in experimental groups and decrease in blood blank of placenta by A-100 ( a one arrowhead ) ( fig2 ) .
FigA2: Microscopic image of control group in 10 yearss placenta with rapid climb A-100, indicates placenta cells and blood blank ( a one caput pointer ) .
Morphology observations on 14-day old placenta
Microscopic observations indicated that placenta of 14 yearss pregnant maternal in experimental group have a less thickness of foetal part bed, and maternal part is more and the addition of cells in placenta maternal and lessening of blood cisterns country in experimental group and control group were seen.
Fig. B3: the alterations of thickness in placenta bed in 14-day old placenta in experimental ( B3 ) and control ( A3 ) groups zoomed A-40 ( two caputs arrow )
Fig. B4: The alterations of the blood cisterns and the figure of placenta part cells in 14-day old placenta in experimental ( B4 ) and control ( A4 ) groups, zoomed A-100 ( one caput pointer )
Consequences of the present survey showed that morphine ingestion during gestation can do hold in the development of embryologic and maternal part of placenta.On pregnant female parent concentration of corticosteron at blood plasma will increase at 2nd period of gestation so we measured corticosteron concentration at 13 yearss after gestation ( 17,18 ) . In understanding with this capable our informations besides indicated the unwritten morphine disposal increased blood plasma corticosteron concentration on habit-forming pregnant female parents ( Tabel 1 ) . Increase in corticosteron denseness of the blood plasma of the pregnant female mice can warrant the side effects in embryonic of these female parents, In add-on the consequences of these surveies were consistent with old 1s, which indicated that cortisol disposal could detain distinction of placental cells. ( 1.5 ) Sing the fact that embryologic development is the consequence of placental natural map and due to the indispensable map of that is the endocrine secernment and material exchange, any upset in normal map of foetal and maternal part of placenta can do abnormalcy on embryo growing ( 3,16,19 ) .
Blood flow is a critical factor on placental map and embryo growing. Morphologic surveies have shown that if physiologic alterations of coiling venas alter during gestation, trophoblast cells attack to placenta and blood flow addition at this site and eventually placental villi disrupted will look other ( 6,20,21 ) . Morphologic and morph metric consequences have shown the affect of unwritten morphia in both embryologic and maternal parts ( Tabel 1 ) . Nowadays, it is stabilized that corticosteron concentration additions in blood of pregnant female parent. besides our survey indicated that unwritten morphia disposal in pregnant mouse caused corticosteron secernment in experimental group. ( 5,17,10 ) Fowden & A ; Forhead declared that exposure of glococorticoides will loss embryo and placenta and this lessen will straight alter the cell rhythm from mitotic to differentiated province ( 5,6,10 ) .Also corticosteron induces proliferation of cytotrophoblast cells of maternal and foetal part of placenta ( 6,22,17 ) by stimulation of procytotrophoblastic cells to shortening of interphase ( 17,5,23 ) , so cells do non hold adequate clip for growing, protein synthesize, reproduction, and adequate growing, and eventually do upset in normal map of placental foetal cells. ( 6,7,23 ) .In contrast lead to late distinction of placental cell and embryologic development ( 5,19,7,10, ) Consequences of present research together with above informations showed high degree of corticosteron by morphine disposal raised maternal part thickness and figure of these cells in placenta of 10th and 14th gestation. In add-on morphia ingestion has decreased more placenta foetal part in contrast to maternal part and this increase cause severs abnormalcy at embryo ( Tabel 1 ) . The old surveies indicated, unwritten morphia disposal in the 9th twenty-four hours of gestation faded nervous tubing and nervous home base development and development of frontal cerebral mantle in embryo were reduced in the seventeenth twenty-four hours of gestation. ( 15,16,24,25 ) The major function of placenta is material exchange between maternal blood and placenta and secretory substances such as steroids, peptides, cytokine, glycoprotein ‘s release from female parent blood and enter to embryo by placenta.In otherwise because ingestion and production of nutrition stuff were determined by placenta any upset in functionality of foetal placental part ( 11,5,6 ) cause hold in development of embryo and placenta. During gestation in parallel with vessel angiogenesis, thickness of foetal placenta part will diminish, so material exchanges become more and eventually morphine consumption additions and lessening in thickness of foetal placenta part is the consequence of this increase ( 20.14,23 ) .So, velocity of morphine passage and riotous effects have direct ratio with lessening of foetal bed thickness. In present research, consequence of morphia resulted lessening in unnatural thickness in embryologic unit of experimental to command group in 10 and 14 twenty-four hours old placenta, and lessening of blood cisterns surface, besides figure of cell in each both 10 and 14 twenty-four hours old placenta in experimental to command group are consistent with addition of corticosteron concentration. ( Tabel 1 ) . The other present researches of morphine consequence on placenta foetal parts of both groups indicated, morphine causes lessening in blood cisterns country and they are consistent with koulin ‘s and Doppler ‘s researches, which the same is true in human.In other researches it was showed, the morphia ingestion orally and injection ally indicated the same consequence ( ,20,26,27 ) . The effectual factors on blood vas contraction are corticosteron and opioides receptors on the membrane of placental cells ( 3,2,7 ) , that located on placental villi and be contracted by morphine stimulation and resulted lessening in blooding, embryologic hypoxia and hold in embryologic development ( 20,28, ) Embryonic part of placenta fundamentally give rise to cyncytiotrophoblast cells and these cells play of import function in embryologic development through secretory map, like estrogen and Lipo-Lutin endocrine ( 10,17 ) and upset in secretory of these cells cause hold eutherian and embryologic development ( 6,19 ) and can worsen embryologic abortion Harmonizing to surveies, morphine disposal caused embryologic abortion and lessening in babes weight in pregnant coneies ( 12 ) . In entire, these consequences indicated unwritten morphia ingestion causes the unusual increase in plasma corticosteron denseness and hold in placenta fetal and maternal part development in Wistar rat but it was non identified if present survey consequences are due to morphine or corticosteron consequence or both of them.Although behavioural upsets in babies or embryologic abortion from habit-forming pregnant female parents need to be studied more.
This survey has been financially sponsored by Research Center of Applied Neuroscience Research Center, Medical University of Baqyiatallah ( a.s. ) .