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1、东北农业大学 硕士学位论文 子宫内膜炎患牛子宫肥大细胞的研究 姓名：刘丽 申请学位级别：硕士 专业：临床兽医学 指导教师：王国卿 20100530 摘 要 I 摘 要 子宫内膜炎是奶牛产后多发的繁殖障碍性疾病，目前对于奶牛子宫内膜炎的研究主要集 中于致病菌分离鉴定、药敏试验和治疗等方面，而对奶牛子宫局部免疫研究较少。肥大细胞 （MC）作为免疫系统中重要效应细胞，对器官局部功能和免疫水平具有重要的调节作用， 研究表明子宫 MC 在发情周期和妊娠过程中均呈规律性变化，说明其是子宫内膜局部状态转 变的重要调节因素之一。而有关奶牛子宫内膜炎时子宫 MC 的结构和功能的研究，还未见报 道。为此，本试验以
2、产后急性化脓性子宫内膜炎患牛为研究对象，通过对血细胞、激素浓度、 血液生化、子宫内膜组胺、SP 和 VIP 含量、HRH1和 HRH2 mRNA 表达量等的检测，及子宫 内膜 MC 显微和超微结构观察，探讨子宫 MC 结构和功能与子宫内膜炎的关系，及 MC 在子 宫局部免疫中的作用。结果表明： 1与健康对照组相比子宫内膜炎组患牛血液白细胞和单核细胞显著增加，而分叶核中 性粒细胞显著减少，表明患牛正处于急性炎症期。 2子宫内膜炎组患牛子宫内膜孕酮浓度为 0.760.28ng/mL，显著高于对照组 0.520.15 ng/mL（p0.05） 。表明子宫收缩受抑制，子宫血液循环障碍，子宫防御机能下降
3、。 3在电镜下，子宫内膜炎组患牛子宫内膜 MC 脱颗粒，并出现空泡。表明子宫内膜炎 引起子宫内膜 MC 释放介质，参与子宫局部防御。 4子宫内膜炎组患牛组胺含量达 80.3054.002ng/mL，显著高于对照组 39.2044.278 ng/mL（p 0.01） ；HRH1 、HRH2 mRNA 表达量为 0.796 0.011、0.926 0.024，也显著高于 对照组 0.6640.004、0.642 0.018（p 0.01） ；而试验组 HRH1 与 HRH2 mRNA 表达量比值显 著降低（0.850.046） ，显著低于对照组 1.0340.002。表明子宫内膜炎时 MC 大量释
4、放组胺， 参与子宫局部炎症反应，HRH1与 HRH2 mRNA 表达量比例失衡，使子宫免疫水平降低。 5子宫内膜炎患组牛子宫内膜、血清中 SP 含量显著升高（p0.01） ，达 1258.06128.88 ng/mL、409.4981.26 ng/mL，而对照组为 308.129.72 ng/mL、59.455.21 ng/mL。表明子宫 内膜炎时神经肽刺激 MC 释放大量组胺，调节子宫免疫。而子宫内膜、血清中 VIP 含量为 615.7370.50 ng/mL、 7.470.37 ng/mL， 显著低于对照组 1667.34153.41 ng/mL、 101.3727.70 ng/mL（p
5、0.01） 。表明子宫内膜炎时，可能存在植物神经功能紊乱。 综上所述，子宫内膜炎时子宫内膜 MC 脱颗粒，释放组胺等介质，参与子宫局部炎症反 应，影响组胺受体表达，引起子宫内膜局部免疫失衡；同时血清和子宫内膜神经肽 VIP 和 SP 浓度变化显著，表明存在植物神经紊乱，VIP 和 SP 在影响子宫局部血液循环的同时，也参与 调解 MC 介质的释放；子宫内膜炎时还表现孕酮水平高和雌激素水平低，子宫收缩受抑制， 子宫内恶漏积聚，利用病菌繁殖。因此 MC 通过释放组胺参与子宫局部免疫，并与神经肽和 卵巢激素密切相关，而在子宫内膜炎中起重要作用；但 MC 介质释放的调解机制有待于进一 步研究。 东北农
6、业大学农学硕士学位论文 II 关键词 奶牛；子宫内膜炎；肥大细胞；结构；组胺；神经肽 Abstract III The Studies Of Uterus MC In Cow Endometeritis Abstract Endometritis is a postnatal reproductive disorder disease in cow. At present, endometritis research for cow has focused on intrauterine flora isolated and appraisement, endometritis medicin
7、e manufacture and treatment, rather on the local immunity of cows less. Mast cells as important effector cells in the immune system, on the organ local function and immunity levels have important regulatory immune. The results show that uterine mast cells in estrous cycle and during pregnancy showed
8、 a regular change. Description mast cells are an important adjustment factor on the endometrial local state change. But when the cow endometritis mast cell structure and function, has not been reported. To this end, the present study of postpartum patients with acute purulent endometritis cows for t
9、he study, by blood cells, hormone levels, blood biochemistry, endometrial histamine, SP and VIP content, HRH1 and HRH2 mRNA expression detection, and endometrial MC microstructure and ultrastructure, research endometritis having an effect on mast cells structure and function, and mast cells in the u
10、terus local immunity role. The results showed as follows: 1. Compared with the healthy control group suffering from cows endometritis group of blood leukocytes and mononuclear cells were significantly increased, while the segmented nuclear neutrophils significantly decreased. This result shows that
11、cows are suffering from acute inflammation. 2. Cows endometritis risk of endometrial progesterone concentration of 0.76 0.28ng/mL, significantly higher than 0.52 0.15 ng / mL (p 0.05). Show that high levels of progesterone and low levels of estrogen, the inhibition of uterine contraction, uterine bl
12、ood circulation disorder, metabolism slows down, the defense function of the uterus decreased. 3. In the electron microscope, endometritis group suffering from cows endometrial MC degranulation and vacuole. This result shows that endometritis endometrium MC release media, participation in the uterus
13、 of local defense. 4. Endometritis cows suffering from histamine content of 80.305 4.002ng/mL, significantly higher than 39.204 4.278 ng / mL (p 0.01), HRH1 and HRH2 mRNA expression level of 0.796 0.011 and 0.926 0.024, also significantly higher than 0.664 0.004 and 0.642 0.018 (p 0.01). But the exp
14、erimental group HRH1 and HRH2 mRNA expression ratio was significantly lower (0.85 0.046), significantly lower than the control group 1.034 0.002. Show that when endometritis a 东北农业大学农学硕士学位论文 IV large number of MC release histamine to participate in the uterus of local inflammatory reaction, histamin
15、e receptor HRH1 and HRH2 mRNA expression imbalance to reduce the level of uterine immunity. 5. Suffering from cows endometritis endometrium, serum SP levels were significantly higher (p 0.01), up to 1258.06 128.88 ng / mL and 409.49 81.26 ng / mL, while the control group was 308.12 9.72 ng / mL and
16、59.45 5.21 ng / mL. That endometritis neuropeptide stimulation MC release large amounts of histamine stimulation, regulation of uterine immunity. The endometrium serum VIP content of 615.73 70.50 ng / mL and 7.47 0.37 ng / mL, significantly lower than the control group 1667.34 153.41 ng / mL and 101.37 27.70 ng / mL (p0.01). That endometritis may exist aut
