口腔矫治器治疗OSAHS对雄性新西兰兔生殖系统的影响

发布时间：2018-06-13 23:00

本文选题：下颌前移矫治器 + 阻塞性睡眠呼吸暂停低通气综合征　；参考：《河北医科大学》2010年硕士论文

【摘要】： 目的:通过建立雄性新西兰兔阻塞性睡眠呼吸暂停低通气综合征(obstructive sleep apnea-hypopnea syndrome,OSAHS)的动物模型,了解OSAHS对生殖系统的影响;采用下颌前移矫治器( Mandibular advancement device, MAD)治疗OSAHS,研究MAD治疗OSAHS对雄兔睾丸光电镜结构及附睾尾中精子的影响。为临床研究MAD治疗OSAHS对男性生殖能力的影响提供实验室数据,为预防儿童OSAHS可能导致的不育提供理论依据。方法:30只雄性新西兰兔随机分为3组:阻塞性睡眠呼吸暂停低通气综合征组(OSAHS组)、下颌前移矫治器治疗组(MAD组)和正常对照组,每组10只。OSAHS组和MAD组实验动物均以1%戊巴比妥钠行全身麻醉,于软腭中间距软硬腭交界0.5cm处,粘膜下肌层注射聚丙烯酰胺水凝胶2ml,动物出现明显打鼾和间断的呼吸暂停,注射后仰卧位头颅侧位片显示软腭后缘上1/4点、1/2点、3/4点处上气道宽度较对照组明显狭窄,动脉血气分析发现注射后动物睡眠发生呼吸暂停时血氧饱和度较对照组降低了22%。以上说明OSAHS动物模型建立成功。MAD组配戴下颌前移矫治器引导下颌向前。三组动物每天以5~6ml/kg经口灌注10%水合氯醛后,仰卧位睡眠4-6小时/天,持续8周。仰卧位睡眠8周后,三组动物在1%戊巴比妥钠全麻下取材,①暴露右侧睾丸于睾丸纵轴、横轴相交处切取1mm3大小的睾丸组织立即放入预冷的4%戊二醛中(此操作在30秒内完成),常规透射电镜标本处理,Epon812包埋,Hitach-7500透射电镜下观察。②取左侧睾丸放入4%多聚甲醛中固定,常规石蜡包埋,HE染色,AX-80万能显微镜下观察睾丸组织学改变。同时,取腭咽部凝胶注射部位组织,固定于4%多聚甲醛中,观察其组织学结构。③同一麻醉状态下,取兔附睾尾组织,置于5ml 37℃生理盐水中尽可能剪碎,混匀后用200目滤网过滤,对滤液进行适当稀释后进行精子计数、精子活力、精子活率及精子畸形率的检测。对上气道间隙、血气分析及附睾尾中精子计数、精子活力、精子活率及精子畸形率的数据结果应用SPSS13.0统计软件进行统计学分析,所有数据以Mean±SD表示,单因素比较采用方差分析,多组间均数两两比较用最小显著性(least significant difference, LSD)检验,相关分析采用直线回归,检验水准为α=0.05。结果:仰卧位上气道间隙宽度分析:OSAHS组软腭1/4点1/2点和3/4点后上气道间隙明显小于MAD组和对照组,差异有统计学意义(P0.05)。MAD组较对照组变窄,但无显著性差异(P0.05)。血气分析:OSAHS组较MAD组和对照组的血氧饱和度氧分压和pH值均显著降低,差别有统计学意义(P0.05)。二氧化碳分压明显高于MAD组和对照组,差别有统计学意义(P0.05)。而MAD组和对照组以上各项指标相比均无统计学意义(P0.05)。注射部位组织学观察可发现:肉眼观察凝胶注射部位,白色透明状,质地均匀。其外有一层薄而透明的包膜,白色凝胶状物质地较韧且富有弹性,局限于软腭软组织中,边界清楚,未见向周围组织扩散。常规HE切片光镜下观察见凝胶被染成蓝色,呈交联状,与周围组织界限清楚,凝胶表面有一层排列整齐的胶原纤维和弹性纤维形成的薄层包膜,没有引起周围组织营养不良及坏死,对周围组织未见明显损伤。肉眼观察三组动物睾丸外形及大小无明显改变。光镜下可以观察到OSAHS组动物睾丸生精细胞的排列较紊乱,生精小管内管腔内容物空虚。生精上皮的层次紊乱,生精小管中可见有细胞呈圆形,胞浆红染,核染色质聚集成团块状。MAD组生精细胞的排列基本正常,部分生精上皮层次紊乱,生精小管中偶见胞浆红染的圆形细胞。正常对照组生精细胞排列整齐,内容物充实,层次分明,细胞未见水肿。透射电镜观察发现,OSAHS组生精细胞中部分线粒体空泡化,稍有肿胀,滑面内质网及粗面内质网扩张,部分粗面内质网脱颗粒,细胞核膜缺损。MAD组细胞未见明显肿胀及空化,内质网轻度扩张、脱颗粒。细胞核膜变化不明显。对照组细胞未见以上变化。附睾尾精子观察发现OSAHS组动物精子计数、精子活力、精子活率较MAD组、对照组明显降低,精子畸形率明显升高,有统计学意义(P 0.05);MAD组动物精子计数、精子活力、精子活率较正常对照组降低,精子畸形率升高,但无统计学意义(P 0.05);将血气分析中血氧饱和度与附睾尾中精子畸形率进行相关分析发现二者呈负相关。结论: 下颌前移矫治器治疗OSAHS,能有效减缓缺氧对睾丸中生精细胞的损伤,有效提高精子质量。
[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.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R766

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