【作者】 浩洪龙；

【导师】 葛利江；

【作者基本信息】 山东农业大学 ， 临床兽医， 2007， 硕士

【摘要】 宫颈是雌性动物维持妊娠和进行分娩的重要器官,在整个妊娠过程中的改变都具有重要意义。宫颈是由上皮、结缔组织、平滑肌和腺体等组成。其中结缔组织占85%~90%,平滑肌占10%~15%。结缔组织包括丰富的胶原纤维、少量的弹性纤维和网状纤维,这些组成的变化与宫颈坚硬程度、宫颈开闭有关。宫颈基质组成的变化与宫颈软化有关。在不同孕期其组织成分比例不同,孕早期以胶原为主,因此宫颈具有一定弹性和韧性,能维持妊娠,分娩开始以前,宫颈必须发生适应分娩的一系列解剖和生理变化,如宫颈组织变软,颈管消失和宫颈扩张以及宫颈中胶原和氨基葡糖减少,含水量增加等,这一过程称为宫颈成熟(cervical ripening)。宫颈成熟是引产成败、产程长短、分娩进行是否顺利的关键。宫颈成熟不仅是一个主动的生化过程,而且是一个炎症反应,有许多介质都参与了宫颈成熟,如孕酮、前列腺素、松弛素、细胞因子及一氧化氮等,但目前关于宫颈成熟过程的控制了解的尚少。试验一山羊分娩期外周血液中激素、细胞因子水平的变化通过检测山羊分娩前后的血清中前列腺素E2(PGE2),孕酮(P4),雌二醇(E2),白细胞介素-1(IL-1)和白细胞介素-8(IL-8)的浓度,研究它们在山羊分娩过程中所发挥的作用和对子宫颈成熟的影响。随机选择健康临近分娩的山羊8只,取分娩前后及分娩过程中的血样共36份,用ELISA检测PGE2,用RIA检测P4,E2,IL-1和IL-8的含量。PGE2、E2、IL-1和IL-8在分娩前血清浓度均逐渐升高,至分娩前一天分别达到481.034pg/mL、60.304pg/mL、0.283ng/mL、0.240ng/mL,分娩后逐渐降低至268.382pg/mL、52.706 pg/mL、0.233ng/mL、0.189ng/mL。其中PGE2在分娩前血清浓度达到最高水平740.964pg/mL,E2在分娩期突然升高至最高水平94.354pg/mL,分娩后则迅速降低。P4在妊娠末期,其血清浓度逐渐下降,至分娩前一天,下降到16.686ng/mL,至分娩时迅速下降为1.871ng/mL。PGE2、P4、E2、IL-1和IL-8在分娩期子宫颈的成熟和扩张过程中均起到重要的作用,其中P4对子宫颈的成熟起到抑制作用,但其在妊娠末期血清浓度的突然下降,则促进了子宫颈的成熟,因此对子宫颈的成熟具有重要的调控作用。试验二山羊分娩期宫颈组织与大网膜及脂肪冰冻切片的观察比较随机选取健康临近分娩的山羊4只进行饲养,在其第二产程期内对其实行剖腹产手术进行样品的采集;也可在分娩出一只胎儿后立刻实行手术采样。分别采取羊子宫颈前、中、后面段组织数块。大网膜及脂肪组织样品可从任一只健康的山羊身上采集。样品采集后,立刻置于装有液氮的液氮罐子中,待冰冻切片机开机预冷12h～24h后,开始切片染色。染色液为苏丹Ⅲ或油红。显微镜下可以观察到,大网膜及脂肪组织切片可见有大量的染成橘红色的脂肪细胞。子宫颈组织切片,在显微镜下亦可观察到有少量的被染成橘红色的脂滴或脂肪细胞。子宫颈组织中的脂滴及脂肪细胞既增加了子宫颈软化的程度,又给子宫颈的扩张提供必要的能量。试验三山羊分娩期宫颈组织电镜切片的制作及观察结果的分析随机选取健康临近分娩的山羊4只,在其第一产程结束后,第二产程开始时或者在第二产程期内,也可在山羊分娩出一只胎儿,宫颈扩张达到最大时,立刻进行手术取样,分别采取羊子宫颈前、中、后段组织数块,所取样品用3.5%的戊二醛进行前固定,然后用PBS水冲洗,再用1%锇酸进行后固定,PBS水冲洗,然后用梯度乙醇脱水,spurr树脂浸透包埋。然后进行切片、染色、观察。分娩期子宫颈电镜图片观察可见,血管中有大量的脂样物质,细胞内有大量的脂肪颗粒,细胞壁破裂,脂肪颗粒释放进入组织。子宫颈组织水肿,线粒体水肿破坏,内有异物生成。电镜切片血管中大量的脂样物质及细胞中脂肪颗粒的释放提示了子宫颈扩张能量的来源,而线粒体的水肿及破坏则说明子宫颈的软化成熟机制。更多还原

【Abstract】 Cervix is an important organ that maintains gravidity and carries out the delivery in the female animal, and its change has an important effect on entire gravidity process. Cervix is composed of epithelium, connective tissue (85%~90%), smooth muscle (10%~15%) and glandular organ etc. Connective tissue contains copious collagen fibers, a little elastic fibers and reticular fibers, and the changes of these fibers are concerned with cervix stiffness degree,cervix opening and shutting. The change of matrix constituent is concerned with cervix softening. The proportion of diverse component is different as advancing pregnancy, for example, the collagen fiber constitute primary element during the early period of gestation, so the cervix with certain elasticity and tenacity can maintain gravidity. A series of the cervical changes in anatomy and physiology appear to adapt delivery prior to parturition,such as the cervical softening,effacement and dilatation, decrease of cervical collagen and glucoseamine, and increase of moisture content and so on, so the process is called cervical ripening. Cervical ripening is the critical process determining the success or failure of inducing labour, the duration of the labour, the easiness of the dilivery.Cervical ripening is not only a active biochemistry process, but also a inflammatory reaction as there are a lot of media have participate in cervical ripening, such as progesterone, prostaglandin, relaxin, cytokine and NO etc., but it is a little known about the regulation to the process of cervical ripening at present. ExperimentⅠ:The change in the level of hormones and cytokines in peripheral blood during the labour in goatsThe serum levels of PGE2, P4, E2, IL-1 and IL-8 during the peripartum period of the goat are determined to study their role in the delivery process and their effect on the cervical ripening.36 seria of goats(n=8) during the peripartum and parturient period are sampled to determine the level of PGE2 by ELISA and P4, E2, IL-1 and IL-8 by RIA .The prepartum serum level of PGE2、E2、IL-1 and IL-8 are gradually rising, one day before parturition the numbers of the parts are 481.034pg/mL、60.304pg/mL、0.283ng/mL、0.240ng/mL, and then gradually decreasing to 268.382pg/mL、52.706 pg/mL、0.233ng/mL、0.189ng/mL after parturition. The level of PGE2 reaches the climax to 740.964pg/mL at the beginning of the parturition, and the level of E2 increases to the climax to 94.354pg/mL during the period of the parturition, and then decrease to the lowest level after parturition; The level of P4 is gradually decreasing during the later period of the parturition, at the day before parturition the level of P4 drop to 16.686ng/mL, and then decrease significantly to 1.871ng/mL.PGE2、P4、E2、IL-1 and IL-8 play important part in the process of the cervix ripening during the pregnant period. Although the P4 can inhibit the maturation, it has enhancement in the maturation of the cervix when P4 is significantly decreasing during the later period of the parturition, so it has an important regulatory function in the maturation of the cervix.ExperimentⅡ:The comparative observation of frozen sections of cervix in labour and nonpregnancy and greater omentum and adipose tissues in goats The healthy goats close to labour (n=8) are at random selected and fed , and the samples at the anterior、midst and posterior part of the cervix are collected on operation at the second stage of labor or collected immediately after the expulsion of a fetus. The samples of greater omentum and fatty tissue can be collected from one of the healthy goats.The samples should be put into the nitrogen container with liquid nitrogen after collection, and can be sliced 12 to 24 hours after the cryoultramicrotome work. The staining solution is SudanⅢor Oil red.A great many of orange-red-stained adipose cells in greater omentum and adipose tissues can be observed under microscope,and a few of orange-red-stained adipose droplets or cells in the cervical tissues are also observed which improve the degree of cervical softening and provide the essential energy for cervical opening. ExperimentⅢ: The preparation and analysis of transmission electron microscope sections of cervix in labour in goatsThe healthy goats close to labour (n=8) are at random selected and fed , and the samples at the anterior、midst and posterior part of the cervix are collected on operation during the second stage of labor or collected immediately after the expulsion of a fetus.The samples should be first fixed with 3.5% glutaraldehyde and washed with PBS, then fixed with osmic acid and washed with PBS, then dehydrated with different gradient of alcohol, finally soaked and embedded with spur resin, sliced, stained and observed.There exist a lot of substance like adipose cells in some blood vessel of cervix and many adipose droplets in cells of cervix in labor under transmission electron microscope, In addition, breakthrough of cell wall, release of adipose droplets into tissues, edema of cervical tissues, rupture of mitochondria and production of foreign body occur.The results above indict that the substance like adipose cells and adipose droplets are energy origin for cervical dilation and edema and rupture of mitochondria is one process for cervical softening and ripening.更多还原