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Objective: The main application of this test HE staining observed after ionizing radiation of primordial follicles, primary follicles, secondary follicles and antral follicle number changes, immunohistochemical methods to detect apoptosis in rat ovarian granulosa cells expression of the situation, and with a P53, of Bcl-2 immune staining method to explore ovarian granulosa cells of P53 and Bcl-2 protein in the expression of the situation, in order to explore the dose of of 0.2 ~~ 1.0 Gy of X-ray ionizing radiation on ovarian morphology and function. Method: 1, the grouping and processing of experimental animals: The experimental animals were 60 rats were randomly divided into six groups, each group of 10 different doses of X-ray irradiation, irradiation voltage 220v target skin distance 100mm, a dose rate of 2.3 Gy / min, the first group of un-irradiated group, storage time equivalent to the irradiation time of the second group, the second group received a radiation absorbed dose of 0.2 Gy, the third group is 0.4 Gy, the fourth group is 0.6 Gy, the fifth group of 0.8Gy, the sixth group to 1.0 Gy. Irradiation for 15 days (three estrous cycle), to give 4% 0.7 ~ 1.5ml of sodium pentobarbital anesthetized by intraperitoneal injection, iodophor disinfection rat back, since the back to cut the skin and muscle tissue into the abdominal cavity, remove both ovaries in a Petri dish with saline washing out after removal of the ovarian fat around. Respectively, with 4% paraformaldehyde and fixed, after 24 hours, embedded, sliced. HE staining method: slices into Harris hematoxylin dye dip after running water, hydrochloric acid, ethanol separations eosin solution and then into the dip water rinse after ethanol differentiation dehydration, solid seal slice in the light microscope follicle count observed primordial follicles, primary follicles, secondary follicles, antral follicle number changes; 3 by TUNEL method: application apoptosis in situ enzyme labeled deoxy nucleic acid, DNA polymerase and terminal transferase (TdT) labeled strand of DNA fragments of the nucleic acid, and then apply the TdT conducted by the end of the reaction, the priority mark early apoptosis cells detect ovarian granulosa cells in the expression of apoptotic bodies; the immunohistochemistry method: slice off wax to the water after rehydration and punch with PBS, antigen retrieval and joined an anti-rinsed with PBS, the addition of two anti-culture SABC method P53 protein staining detected in granulosa cells and expression of Bcl-2 protein situation. Five pairs of immunohistochemistry graphic image optical density value and the gray value calculated, and analyzed statistically. Results: (1) X-ray ionizing radiation (0.2Gy ~ 1.0Gy) enables primordial follicles, primary follicles, the decrease in the number of secondary follicles (P lt; 0.01), reduced to a primordial follicles. The number of antral follicles after different doses of X-ray irradiation did not change significantly with the sham irradiation group (P gt; 0.05). Absorbent capacity of 0.4Gy ~ 1.0Gy ovarian granulosa cells in the presence of nuclear vacuoles. (2) The absorbed dose 0.2Gy ovarian granulosa cells, p53 protein levels lower than those in the sham irradiation group (P lt; 0.01), Bcl-2 protein expression levels higher than the sham irradiation group (P lt; 0.05). (3) absorbed dose of 0.4 ~ 1.0 Gy, the level of expression of P53 protein in ovarian granulosa cells was significantly higher than that in the sham irradiation group (P lt; 0.01), and the expression level increased with the increase of the absorbed dose; Bcl-2 protein expression significantly lower than those in the sham irradiation group (P lt; 0.01), and decreased with the increase in the expression level of the absorbed dose. Was significantly higher than that (4) absorbed dose of 0.2 Gy, ovarian granulosa cell apoptosis amount lower than the sham-irradiated group (P lt; 0.01); absorbed dose of 0.4 ~ 1.0 Gy, the level of expression of apoptotic bodies. sham irradiation group, and also increased with increased expression of the absorbed dose (P lt; 0.01). Conclusion: The dose between 0.4 ~ 1.0 Gy of ionizing radiation can increase in ovarian granulosa cell apoptosis, and reduce the number of primordial follicles, primary follicles, secondary follicles, which ovarian damage, such as the cause of some of the ovarian cell damage it will affect the ability to conceive in the future, even if the pregnancy is the quality of the embryonic development there is a certain risk, as all injury occurs, may result in premature ovarian failure, affecting the functional regulation of gonadal hormone secretion. Absorbed dose in the clinical, from small to large order of general radiographic examination, X-ray, CT examination. The minimum dose of 2.31 ~ 5.35 mGy, the maximum dose of 4 Gy. From the experimental results, when the absorbed dose equal to the the 0.2Gy has caused the reduction of the number of follicles, granulosa cell apoptosis can lead to the absorbed dose is greater than 0.4Gy radiological examination, pelvic and other vital organs to make The protection is imperative.
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