本文選題：地塞米松　切入點(diǎn)：腭裂　出處：《吉林大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景：腭裂是我們?cè)谂R床工作中最常見(jiàn)的顱面先天畸形之一，現(xiàn)如今已被認(rèn)識(shí)到可能是由多種母親的某些基因及所生活的環(huán)境因素所導(dǎo)致的，在世界各地每出生500至2500名新生兒中就有1名患兒出現(xiàn)唇腭裂，致病率達(dá)到0,2%-0.04%，這給患兒在日常的進(jìn)食、與人交流等基本生活方面帶來(lái)障礙，還會(huì)在患兒成長(zhǎng)過(guò)程中帶來(lái)極大的心理影響。在近20年的研究報(bào)告顯示地塞米松是一種導(dǎo)致胚胎畸形較強(qiáng)的藥物，但究竟是一次性大劑量給藥產(chǎn)生畸形率高還是小劑量多次給藥所產(chǎn)生的畸形率高并沒(méi)有明確的解釋。許多國(guó)內(nèi)外學(xué)者在研究中發(fā)現(xiàn)地塞米松主要作用于腭突間充質(zhì)細(xì)胞，在腭突發(fā)育過(guò)程中可抑制EPM細(xì)胞的增殖，但細(xì)胞的凋亡是否也與腭裂的形成存在某種關(guān)系尚不清楚，所以本實(shí)驗(yàn)通過(guò)建立動(dòng)物模型觀察不同劑量地塞米松誘導(dǎo)小鼠致畸率的高低及與細(xì)胞凋亡的關(guān)系。 實(shí)驗(yàn)?zāi)康模禾接懶∈箅窳雅c不同劑量地塞米松的關(guān)系及在腭裂形成過(guò)程中與BMP-2和細(xì)胞凋亡的相關(guān)性。 模型的建立和方法：隨機(jī)選取生活環(huán)境相近并且健康的近交系小鼠，雌雄比例按2:1于晚18:00合籠飼養(yǎng)，所有小鼠均在光照及黑暗時(shí)間12h的同樣環(huán)境下飼養(yǎng)。并于次日早7:30檢查雌性小鼠的陰栓，見(jiàn)陰栓之日起為孕0d（0GD），并依次稱體重作標(biāo)記，于10GD上午7:30再依次稱體重，若體重顯著增加2g以上者可認(rèn)為妊娠陽(yáng)性。并取妊娠陽(yáng)性小鼠隨機(jī)選取45只分為三組，每組15只放于同籠飼養(yǎng)并做好標(biāo)記分別為小劑量組于妊娠后11.5d、12.5d、13.5d連續(xù)三天腹腔注射6mg/kg地塞米松；大劑量組于妊娠后11.5d腹腔內(nèi)一次性大劑量注射50mg/kg地塞米松；空白對(duì)照組孕鼠在正常情況下飼養(yǎng)，不給予任何特殊處理。所有小組均在14.5d以脫臼法依次處死，每組小孕鼠均在處死后立即放置于冰塊上，并剖腹取出胎鼠，并觀察各組孕鼠的上顎的大體形態(tài)及常規(guī)HE染色病理切片、免疫組化、透射電鏡掃描。 結(jié)果：各組小鼠在飼養(yǎng)過(guò)程中均未出現(xiàn)疾病或死亡現(xiàn)象。經(jīng)過(guò)肉眼觀察可得出小劑量組共懷孕175只孕鼠，其中發(fā)生腭裂胎鼠72只，未發(fā)生腭裂胎鼠103只，平均體重是0.31克，腭裂發(fā)生率是41.14%；大劑量組共懷孕103只，其中發(fā)生腭裂的胎鼠63只，未發(fā)生腭裂的胎鼠40只，平均體重是0.43克，腭裂發(fā)生率是61.17%；空白對(duì)照組共懷孕139只，其中發(fā)生腭裂的胎鼠是7只，平均體重是0.54克，腭裂發(fā)生率是5.04%，三組小鼠稱重后之間進(jìn)行統(tǒng)計(jì)學(xué)分析小鼠平均體重和腭裂發(fā)生率P 0.05，有統(tǒng)計(jì)學(xué)差異。經(jīng)HE染色光鏡下可看到腭裂胎鼠的上腭正中位置有一條縱行走向邊緣不規(guī)整的裂隙，分析后可見(jiàn)大劑量組腭裂發(fā)生率明顯高于小劑量組；經(jīng)過(guò)TUNEL染色觀察腭裂小鼠腭突出現(xiàn)大量凋亡細(xì)胞，經(jīng)免疫組化觀察三組BMP-2變化可看出不同劑量之間并沒(méi)有明顯差異，但表達(dá)情況均較空白對(duì)照組表達(dá)減少，并且在透射電鏡下可見(jiàn)腭裂胎鼠的頭部的腭突間充質(zhì)細(xì)胞形態(tài)及細(xì)胞器結(jié)構(gòu)與空白對(duì)照組明顯不同。 結(jié)論： 1.一次大劑量注射地塞米松較多次少量注射地塞米松更易導(dǎo)致腭裂的發(fā)生。 2.劑量依賴性的腭裂發(fā)生率變化與腭突間充質(zhì)細(xì)胞的凋亡水平密切相關(guān) 3.在地塞米松引起的腭裂胎鼠腭突處，BMP-2的表達(dá)較正常發(fā)育的小鼠明顯減少。 4.不同劑量地塞米松引起的腭裂腭突的BMP-2的表達(dá)未見(jiàn)明顯差異。
[Abstract]:Background: cleft palate is the most common in the clinical work of the craniofacial congenital malformation of now has been recognized to be caused by a variety of genes and mother living environment factors, born in the world every 500 to 2500 births in 1 children with cleft lip and palate appear, pathogenicity the rate of up to 0,2%-0.04%, this to the children in the daily diet, and communicate the basic life difficult, but also bring great psychological impact on children in the growth process. In the nearly 20 years of research report that dexamethasone is a cause of fetal abnormalities strong drugs, but whether it is a single large dose of administration produce deformity rate high or low dose of medicine repeatedly to the deformity rate produced by high and there is no clear explanation. Many domestic and foreign scholars have found that dexamethasone acts mainly on palatal mesenchymal cells in the study, in the palatal hair The proliferation of EPM cells can be inhibited during the course of growth. However, whether the cell apoptosis is related to the formation of cleft palate is not clear. Therefore, the animal model was established to observe the relationship between the teratogenic rate and the apoptosis of mice induced by different doses of dexamethasone.
Objective: To investigate the relationship between cleft palate and different doses of dexamethasone and the correlation between BMP-2 and apoptosis during the formation of cleft palate.
The establishment of the model and method: randomly selected similar living environment and healthy inbred mice, male and female ratio of 2:1 in the late 18:00 cages, all the mice were raised in the same environment light and dark time 12h. And on the next day at 7:30 in the morning to check the vaginal plug in female mice, vaginal plug date pregnant 0d (0GD), and in turn the body for markers in 10GD at 7:30 in the morning and then weighed, if the body weight increased significantly more than 2G can be considered positive and positive pregnancy. Pregnant mice were randomly selected 45 rabbits were divided into three groups, 15 rats in each group to put in the same cage and make mark were small dose to pregnant 11.5d, 12.5d, 13.5d for three consecutive days intraperitoneal injection of 6mg/kg dexamethasone; high dose group in pregnancy after 11.5d intraperitoneal single dose injection of 50mg/kg dexamethasone; control group of pregnant rats reared under normal conditions, without any special treatment. All groups were killed in the order of 14.5d at the time of dislocations. Every group of small pregnant mice was placed on ice on the ice immediately after they were executed, and the fetuses were taken out of the abdominal cavity. The gross morphology of the upper jaw and routine HE staining pathological sections were observed, and immunohistochemistry and transmission electron microscope scanning were used.
Results: the mice in the breeding process were not found in the phenomenon of disease or death was observed by naked eyes. The small dose group were pregnant and 175 pregnant rats, the occurrence of cleft palate 72 rats, 103 rats without cleft palate, the average weight is 0.31 grams, the incidence of cleft palate is 41.14%; the large dose group were pregnant 103, the occurrence of fetal cleft palate 63 rats, without the occurrence of cleft palate in fetal rats was 40, the average weight is 0.43 grams, the incidence of cleft palate was 61.17%; control group were pregnant and 139, which occurred in cleft palate of fetal rats is 7, the average weight is 0.54 grams, the incidence of cleft palate was 5.04%, three mice weighing statistical analysis of incidence rate of P 0.05 and the average weight of the mice after cleft palate, there were significant differences. After HE staining under light microscope can see the fetal palate cleft palate with a longitudinal center position toward the edge of irregular fracture analysis, visible after high dose group of cleft palate The incidence rate was significantly higher than the low dose group; after TUNEL staining of palatal cleft palate mice appear a large number of apoptotic cells by immunohistochemical observation of three groups of BMP-2 changes can be seen and there is no significant difference between different dose, but the expression was compared with blank control group was decreased, and the transmission electron microscope of cleft palate mice head palate process of mesenchymal cell morphology and cell structure and blank control group were significantly different.
Conclusion:
1. a large dose of dexamethasone injected with dexamethasone a few times is more likely to cause cleft palate.
The changes in the incidence of 2. dose dependent cleft palate are closely related to the apoptosis level of the palatine interprocess mesenchymal cells
3. in the palatine process of the cleft palate mouse induced by dexamethasone, the expression of BMP-2 was significantly lower than that in the normal developing mice.
4. there was no significant difference in the expression of BMP-2 in cleft palate and palate process caused by different doses of dexamethasone.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R782.22

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