Effects On Ovary Structure And Cell Apoptosis Of Different Range Hysterectomy

Objective:Through establishing animal models,explore the influence on ovarian follicular,Cellular form and apoptosis after different range hysterectomies.And to explore the influence mechanism on ovarian function after a hysterectomy.Add basis for clinical guidance to choose proper operation mode.Methods:1.Experimental animal models and materials:There are 36 adult female and counterparts New Zealand rabbits,randomly divided into three groups,each group of 12,respectively is the control group(groupA),triangle hysterectomy group(group B) and total hysterectomy group(Group C).Make different range hysterectomy according to the group.Group A:the uterus and ovaries should be exposed half an hour,and don't make any operation,then close the abdominal cavity.All the experimental rabbits were raised 4 months after the surgery.Determine the estrogen cycle through observing the vaginal exfoliated cells.Then obtain the materials.Make the rabbits anesthesia and fixed,Looking for the bilateral ovaries and resecting them after into the abdominal cavity.When the left ovary was resected,get part of the organization which near the ovarian hover ligament immediately,and cut it into 1x1x4mm~3 organization,then put into 4%glutaraldehyde fixation solution,prepare for electron microscopy.Fix the remaining left ovarian tissue with 70% alcohol solution prepare for FCM.Right ovary,using formaldehyde solution fixed,prepared for light microscopy.2.Measuring method of each index:manufacture the ovaries sample which fixing in 70%alcohol into monodisperse cell suspension and detect the rate of apoptosis by flow cytometry.Afer alcoholic dehydrating right ovary which fixing by formaldehyde solution in normal gradient,transparenting by dimethyl benzene,paraffin imbedding,slicing and HE dyeing, observing by light microscope.Make the left tissue of ovaries into sample by fixing and embedding,After locating by semithin section,we can observe the fine structure of the ovaries granular cell.Measurement data results are express in (mean±standard deviation),and all the data is dealt with statistics software of SPSS13.0 containing test t and analysis of variance.α=0.05,P<0.05.Results:1.Test results of apoptosis rate of oarian cells through the flow cytometry determination,and the single factor analysis of variance confirmed:the oarian cells' apoptosis rate of the total hysterectomy group(22.48±5.13%) is higher than the control group(6.91±3.48%),the differences have a statistically significance(P<0.05).The oarian cells' apoptosis rate of the total hysterectomy group is higher than triangle hysterectomy group(7.20±3.18%),the differences have a statistically significance(P＜0.05).The differences between the triangle hysterectomy group and the control group are no statistical significance.2.Observe the form of granular cells and fine structure by electron microscope.Group A:The permutation of granular cells are normal and nature.The density of the nucleus and the matrix are uniform.Cells show anomalous ellipse,caryon is large and dyeing light,there are one or two reticular nucleus fruit-kernel,there are many cell organ.Crista mitochondriales arrages regulatly and shows tubiform,the construction is common.There can be seen lots of obesity in the endochylema.Cell jions compact,In the granular cells which near to the zona pellucida we can find that ecphyma linked with it.Group B:It is basically the same display as Group AGroup C:The quantity of the cell organ in granular cell is reduce.The Sum of the smooth endoplasmic reticulum decrease and so does the ridge count.Crista mitochondriales shows plate shape,construction is unclear.Rough endoplasmic reticulum expand lightly.There are more vacuolus can be seen in the periplast,periplast dropsy partly,there is space-occupying lesion in caryotheca partly.Perinuclear space can be seen stretched lightly.Coloboma in the cell envelope of the granular cell also can be seen.Caryon dropsy partly,low electro-dense can be seen, vascular pool of the cells widen.the distance between granular cell and zona pellucida become large and ecphyma decrease.3.Observe the organization of ovaries and developmental condition of follicular development by light microscope.Group A:There are a lot of primitive follicles in the superficial department of ovarian cortex.There are follicles of different developmental stages in the cortex,including primary follicles,subprime follicles,and mature follicles.Most oocytes of the growth of follicles are large,and they have fine particulate chromatin,their mesh nucleolus are visible.The folliculars'morphology rules,their sizes are different and they have sinus formation.There are corpus luteum,interstitial structure and closure of follicles in the cortex.Group B:It is basically the same as Group A under light microscope.Group C:The sum of the folliculars in ovarian cortex are reduce,especially growth of follicles and mature follicles.There are many closure of follicles.Most folliculars'form are irregular.The nuclear of oocytes are deviation,pyknotic and the zona pellucida of oocytes are collapse.The connect between granular cells are loose,they are atrophy and fall off into follicular cavities.There are a lot of Fibrosis in the interstitial organization of ovarian tissue.Conclusions:1.After the total hysterectomy,the apoptosis of ovarian cells are increasing.2.After the total hysterectomy,there are morphological changes and organelles structure damage to some ovarian granulosa cells.There are a series of apoptosis performance.3.After the total hysterectomy,folliculars of ovarian tissue are dysplasia,there are a series of blood anoxic change.4.Triangle hysterectomy surgery are no obvious effect on ovarian tissue and granular cells form.It maybe the morphology basic of the lack of ovarian function after hysterectomy.We should consider the scope of operation fully and master the surgical indications strictly.