發(fā)布時間：2018-06-13 23:00

  本文選題：下頜前移矯治器 + 阻塞性睡眠呼吸暫停低通氣綜合征　； 參考：《河北醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的:通過建立雄性新西蘭兔阻塞性睡眠呼吸暫停低通氣綜合征(obstructive sleep apnea-hypopnea syndrome,OSAHS)的動物模型,了解OSAHS對生殖系統(tǒng)的影響;采用下頜前移矯治器( Mandibular advancement device, MAD)治療OSAHS,研究MAD治療OSAHS對雄兔睪丸光電鏡結(jié)構(gòu)及附睪尾中精子的影響。為臨床研究MAD治療OSAHS對男性生殖能力的影響提供實驗室數(shù)據(jù),為預(yù)防兒童OSAHS可能導(dǎo)致的不育提供理論依據(jù)。 方法:30只雄性新西蘭兔隨機分為3組:阻塞性睡眠呼吸暫停低通氣綜合征組(OSAHS組)、下頜前移矯治器治療組(MAD組)和正常對照組,每組10只。OSAHS組和MAD組實驗動物均以1%戊巴比妥鈉行全身麻醉,于軟腭中間距軟硬腭交界0.5cm處,粘膜下肌層注射聚丙烯酰胺水凝膠2ml,動物出現(xiàn)明顯打鼾和間斷的呼吸暫停,注射后仰臥位頭顱側(cè)位片顯示軟腭后緣上1/4點、1/2點、3/4點處上氣道寬度較對照組明顯狹窄,動脈血氣分析發(fā)現(xiàn)注射后動物睡眠發(fā)生呼吸暫停時血氧飽和度較對照組降低了22%。以上說明OSAHS動物模型建立成功。MAD組配戴下頜前移矯治器引導(dǎo)下頜向前。 三組動物每天以5~6ml/kg經(jīng)口灌注10%水合氯醛后,仰臥位睡眠4-6小時/天,持續(xù)8周。仰臥位睡眠8周后,三組動物在1%戊巴比妥鈉全麻下取材,①暴露右側(cè)睪丸于睪丸縱軸、橫軸相交處切取1mm3大小的睪丸組織立即放入預(yù)冷的4%戊二醛中(此操作在30秒內(nèi)完成),常規(guī)透射電鏡標(biāo)本處理,Epon812包埋,Hitach-7500透射電鏡下觀察。②取左側(cè)睪丸放入4%多聚甲醛中固定,常規(guī)石蠟包埋,HE染色,AX-80萬能顯微鏡下觀察睪丸組織學(xué)改變。同時,取腭咽部凝膠注射部位組織,固定于4%多聚甲醛中,觀察其組織學(xué)結(jié)構(gòu)。③同一麻醉狀態(tài)下,取兔附睪尾組織,置于5ml 37℃生理鹽水中盡可能剪碎,混勻后用200目濾網(wǎng)過濾,對濾液進(jìn)行適當(dāng)稀釋后進(jìn)行精子計數(shù)、精子活力、精子活率及精子畸形率的檢測。 對上氣道間隙、血氣分析及附睪尾中精子計數(shù)、精子活力、精子活率及精子畸形率的數(shù)據(jù)結(jié)果應(yīng)用SPSS13.0統(tǒng)計軟件進(jìn)行統(tǒng)計學(xué)分析,所有數(shù)據(jù)以Mean±SD表示,單因素比較采用方差分析,多組間均數(shù)兩兩比較用最小顯著性(least significant difference, LSD)檢驗,相關(guān)分析采用直線回歸,檢驗水準(zhǔn)為α=0.05。 結(jié)果:仰臥位上氣道間隙寬度分析:OSAHS組軟腭1/4點1/2點和3/4點后上氣道間隙明顯小于MAD組和對照組,差異有統(tǒng)計學(xué)意義(P0.05)。MAD組較對照組變窄,但無顯著性差異(P0.05)。血氣分析:OSAHS組較MAD組和對照組的血氧飽和度氧分壓和pH值均顯著降低,差別有統(tǒng)計學(xué)意義(P0.05)。二氧化碳分壓明顯高于MAD組和對照組,差別有統(tǒng)計學(xué)意義(P0.05)。而MAD組和對照組以上各項指標(biāo)相比均無統(tǒng)計學(xué)意義(P0.05)。 注射部位組織學(xué)觀察可發(fā)現(xiàn):肉眼觀察凝膠注射部位,白色透明狀,質(zhì)地均勻。其外有一層薄而透明的包膜,白色凝膠狀物質(zhì)地較韌且富有彈性,局限于軟腭軟組織中,邊界清楚,未見向周圍組織擴散。常規(guī)HE切片光鏡下觀察見凝膠被染成藍(lán)色,呈交聯(lián)狀,與周圍組織界限清楚,凝膠表面有一層排列整齊的膠原纖維和彈性纖維形成的薄層包膜,沒有引起周圍組織營養(yǎng)不良及壞死,對周圍組織未見明顯損傷。 肉眼觀察三組動物睪丸外形及大小無明顯改變。光鏡下可以觀察到OSAHS組動物睪丸生精細(xì)胞的排列較紊亂,生精小管內(nèi)管腔內(nèi)容物空虛。生精上皮的層次紊亂,生精小管中可見有細(xì)胞呈圓形,胞漿紅染,核染色質(zhì)聚集成團(tuán)塊狀。MAD組生精細(xì)胞的排列基本正常,部分生精上皮層次紊亂,生精小管中偶見胞漿紅染的圓形細(xì)胞。正常對照組生精細(xì)胞排列整齊,內(nèi)容物充實,層次分明,細(xì)胞未見水腫。 透射電鏡觀察發(fā)現(xiàn),OSAHS組生精細(xì)胞中部分線粒體空泡化,稍有腫脹,滑面內(nèi)質(zhì)網(wǎng)及粗面內(nèi)質(zhì)網(wǎng)擴張,部分粗面內(nèi)質(zhì)網(wǎng)脫顆粒,細(xì)胞核膜缺損。MAD組細(xì)胞未見明顯腫脹及空化,內(nèi)質(zhì)網(wǎng)輕度擴張、脫顆粒。細(xì)胞核膜變化不明顯。對照組細(xì)胞未見以上變化。 附睪尾精子觀察發(fā)現(xiàn)OSAHS組動物精子計數(shù)、精子活力、精子活率較MAD組、對照組明顯降低,精子畸形率明顯升高,有統(tǒng)計學(xué)意義(P 0.05);MAD組動物精子計數(shù)、精子活力、精子活率較正常對照組降低,精子畸形率升高,但無統(tǒng)計學(xué)意義(P 0.05);將血氣分析中血氧飽和度與附睪尾中精子畸形率進(jìn)行相關(guān)分析發(fā)現(xiàn)二者呈負(fù)相關(guān)。 結(jié)論: 下頜前移矯治器治療OSAHS,能有效減緩缺氧對睪丸中生精細(xì)胞的損傷,有效提高精子質(zhì)量。
[Abstract]:Objective: to understand the effect of OSAHS on the reproductive system by establishing the animal model of obstructive sleep apnea-hypopnea syndrome (OSAHS) in male New Zealand rabbits, and to study the testicular light of male rabbits by using the mandibular forward moving appliance (Mandibular advancement device, MAD) for the OSAHS. The effects of electron microscopic structure and sperm in the epididymal tail provide laboratory data for the clinical study of the effects of the MAD treatment of OSAHS on male reproductive ability, and provide a theoretical basis for preventing the possible infertility caused by OSAHS in children.
Methods: 30 male New Zealand rabbits were randomly divided into 3 groups: obstructive sleep apnea hypopnea syndrome group (group OSAHS), mandibular anterior shift appliance group (group MAD) and normal control group, 10.OSAHS groups and MAD groups in each group were treated with 1% pentobarbital sodium, at the middle distance from the soft palate to the soft and hard palate at 0.5cm, and the submucous muscle. After injection of polyacrylamide hydrogel 2ml, the animals had obvious snoring and intermittent apnea, and the lateral position of the supine position after injection showed 1/4 point in the posterior margin of the soft palate, 1/2 point, and the upper airway width at the 3/4 point was obviously narrower than the control group. The arterial blood gas analysis found that the blood oxygen saturation was lower when the apnea was suspended in the animal after injection. Lower 22%. above indicated that the OSAHS animal model was successfully established and the.MAD group wore the mandibular advancement appliance to guide the mandible forward.
After three groups of animals were perfused with 10% chloral chloral with 5~6ml/kg every day, sleeping in the supine position was 4-6 hours per day for 8 weeks. After sleeping in the supine position for 8 weeks, the three groups of animals were harvested under 1% pentobarbital sodium general anesthesia, and the right testicles were exposed to the longitudinal axis of the testis, and the 1mm3 size of the testis was cut into the precooled 4% glutaraldehyde immediately. In 30 seconds), the routine transmission electron microscope specimen treatment, Epon812 embedding and Hitach-7500 transmission electron microscopy were observed. 2. The left testis was fixed in 4% polyformaldehyde, conventional paraffin embedding, HE staining, and AX-80 omnipotent microscope observed the histological changes of the testis. At the same time, the tissue of the injection site of the palatopharynx gel was taken in 4% polyformaldehyde. Under the same anesthetic state, the rabbit epididymal tail tissue was taken to be cut in 5ml 37 degrees centigrade saline, and then filtered with 200 mesh filter. After proper dilution, the sperm count, sperm motility, sperm motility and sperm abnormality rate were measured.
The data of the upper airway space, the blood gas analysis and the sperm count in the epididymis, sperm motility, sperm motility and sperm abnormality were statistically analyzed with SPSS13.0 software. All the data were expressed in Mean + SD, and the single factor was compared with the variance analysis. The minimum significant (least significant differ) was compared between the multiple groups (least significant differ) Ence (LSD) test, correlation analysis using linear regression, the test level is =0.05.
Results: the width of the upper airway gap in the supine position: the 1/4 point of the soft palate in the OSAHS group? 1/2 point and the 3/4 point in the upper airway gap was significantly smaller than that in the MAD group and the control group; the difference was statistically significant (P0.05) in the.MAD group, but there was no significant difference (P0.05). The blood gas analysis of the OSAHS group was compared with the oxygen saturation, the oxygen partial pressure and the pH value of the MAD and the control groups. The difference was statistically significant (P0.05). The partial pressure of carbon dioxide was significantly higher than that of the MAD group and the control group, and the difference was statistically significant (P0.05), but there was no significant difference between the MAD group and the control group (P0.05).
The histological observation of the injection site showed that the injection site of the gels was observed by the naked eye, white transparent and uniform, and there was a thin and transparent coating on the outside, and the white gelatinous substance was toughened and elastic, limited to soft palate soft tissue, the boundary was clear, and the surrounding group was not spread. The gel was stained blue under conventional HE microscope. The color is cross linked, clearly defined with the surrounding tissue. The surface of the gel has a layer of thin layers of collagen fibers and elastic fibers. It does not cause malnutrition and necrosis of the surrounding tissue and has no obvious damage to the surrounding tissue.
There was no obvious change in the shape and size of the testicles in the three groups of animals. Under the light microscope, the arrangement of spermatogenic cells in the testis of the OSAHS group was disordered. The contents of the lumen in the seminiferous tubules were empty. The level of the spermatogenic epithelium was disordered. The cells in the seminiferous tubules were round, the cytoplasm red dye, and the nuclear chromatin aggregated and aggregated in group.MAD. The arrangement of the cells was basically normal, some of the spermatogenic epithelium was in disorder, and the circular cells of the cytoplasm red dye were seen in the seminiferous tubules. The normal control group was arranged neatly, the contents were full, the level was distinct, and the cells did not have edema.
Transmission electron microscopy showed that some mitochondria were vacuolated in the spermatogenic cells of OSAHS group, slightly swelling, the endoplasmic reticulum and endoplasmic reticulum dilated, some rough endoplasmic reticulum degranulation, and no obvious swelling and cavitation in the.MAD group of the cell nuclear membrane defect, the endoplasmic reticulum was slightly dilated and degranulation. The cell nuclear membrane was not changed obviously. The cells of the control group were not seen. The cells of the control group did not see the cells. The above changes.
The observation of epididymal sperm found that the sperm count, sperm motility and sperm viability of the OSAHS group were lower than that of the MAD group, the control group was significantly lower, the sperm malformation rate was significantly increased (P 0.05); the sperm count, sperm motility, sperm viability of the MAD group were lower than the normal control group, and the sperm abnormality rate increased, but there was no statistical significance (P 0.05); the blood was not statistically significant (P 0.05). Correlation analysis between blood oxygen saturation and sperm deformity rate in epididymal tail showed that there was a negative correlation between the two parameters.
Conclusion:
Treatment of OSAHS with mandibular advancement appliance can effectively alleviate the damage of hypoxia to spermatogenic cells in testis and effectively improve sperm quality.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R766

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8 王軼娜;慢性間歇低氧通過TNF-α調(diào)控fractalkine在OSAHS所致肝損傷中的作用[D];中南大學(xué);2012年

9 王宇;阻塞性睡眠呼吸暫停低通氣綜合征的一期多平面手術(shù)治療與血清標(biāo)記物變化的研究[D];中國人民解放軍軍醫(yī)進(jìn)修學(xué)院;2010年

10 于興娟;啄治法治療兒童鼾眠的臨床療效評價研究[D];山東中醫(yī)藥大學(xué);2010年

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本文編號：2015843