多发性硬化导致的膀胱功能障碍以及雌激素对其治疗作用的研究

发布时间：2018-03-04 00:11

本文选题：多发性硬化　切入点：神经源性膀胱　出处：《郑州大学》2017年硕士论文　论文类型：学位论文

【摘要】：背景和目的多发性硬化导致的膀胱功能障碍是泌尿系统里面极为复杂的疾病。多发性硬化会给中枢神经系统带来多样的病理变化,与此同时会出现各个系统临床症状。对于泌尿系统,由于受损的泌尿系功能相关的神经病变部位以及受损伤程度不同也会导致泌尿系统功能也将出现各种变化,其中膀胱功能障碍主要表现为尿频、尿急、尿潴留、尿失禁等。目前对于多发性硬化这个疾病本身的研究较多,而关于多发性硬化神经源性膀胱研究则较少.治疗方法上面也是主要以控制症状,改善泌尿系统功能为主,比如间歇导尿、抗胆碱药物、肉毒素、辣椒素、手术等等,治疗效果并不理想,针对多发硬化的治疗如免疫抑制剂、免疫球蛋白注射、血浆置换以及β-干扰素等效果也不能让人们完全满意,因此需要一种新的同时针对多发性硬化和神经源性膀胱的治疗方法。近期有研究表明男女性别的差异会造成自身免疫性疾病发病差别,女性患者多发性硬化的患者是男性的2倍,女性怀孕后期多发性硬化发病率降低到原来的20%,有研究表明雌性激素对多发性硬化有防止疾病进一步恶化,促进神经修复的作用。另外还有报道证明雌激素受体在膀胱的分布,以及雌激素对女性膀胱结构和功能的影响,提示了雌激素在治疗多发性硬化神经源性膀胱的潜在可能。本实验通过实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的动物模型观察多发性硬化导致的膀胱功能障碍,并通过给予模型动物适量的雌激素处理,观察模型动物变化情况,初步验证雌激素在多发性硬化中神经源性膀胱的可能治疗作用。实验方法1建立EAE小鼠模型:8-13周的C57BL/6雌性小鼠,应用200ug mog35-55,与等体积含CFA量为10mg/ml的完全弗氏佐剂混匀,完全乳化成乳剂,皮下两点注射,于免疫当天及免疫后48小时腹腔注射400ng百日咳毒素诱导EAE小鼠模型。自免疫当天起,观察并记录大鼠体重变化、临床表现及神经功能评分。2分组以及给药:选取雌性C57BL6小鼠30只,随机分10只对照组单纯给予完全费氏佐剂以及百日咳,10只为EAE模型组,10只为雌激素处理组,即在完成免疫处理后每日给予雌激素(50μmol/L)100μl皮下注射。3标本采集:在试验23天时,将小鼠用10%水合氯醛麻醉后,取出膀胱组织,称重,放入液氮中保存,然后取多聚甲醛进行全身灌流,取出小鼠脊髓组织用于制作石蜡切片。4相关指标检测:Luxol fast blue(LFB)染色和苏木素-伊红(HE)染色观察脊髓炎性细胞浸润及脱髓鞘情况,用Western-Blot检测膀胱组织中雌激素受体蛋白表达变化情况。结果1 EAE模型组的小鼠:神经功能临床评分、体重、以及泌尿系统功能变化都具有典型的MS临床表现,HE染色表现出较明显相关的炎性细胞浸润。LFB染色表现出了明显的脊髓脱髓鞘现象。2在多发性硬化神经源性膀胱小鼠膀胱组织中,相对于对照组小鼠膀胱中雌激素受体明显降低,给予雌激素治疗后,雌激素受体表达略微上调。3在给予EAE小鼠雌激素治疗后,小鼠发病率降低,症状严重程度较未特殊处理的EAE小鼠轻,平均神经功能评分降低。结论1 EAE神经系统临床症状的评分与多发硬化性神经源膀胱的症状密切相关。2雌激素以及雌激素受体在多发硬化性神经源膀胱疾病中发生可能起到重要用。3在对EAE模型小鼠进行雌激素处理后,发病率以及症状严重程度相对于未经雌激素处理的模型组明显降低,表明雌激素对多发性硬化导致神经源性膀胱有明显的改善作用。
[Abstract]:Background and objective: multiple sclerosis induced bladder dysfunction is the urinary system which is extremely complicated diseases. Multiple sclerosis will bring various pathological changes to the central nervous system, at the same time the system will appear in various clinical symptoms. For the urinary system, urinary system dysfunction due to the related nerve lesion and damage degree will also cause urinary tract function will also appear various changes, including bladder dysfunction mainly for frequent micturition, urgency of urination, urinary retention, urinary incontinence and so on. At present there are many studies on multiple sclerosis of the disease itself, and on multiple sclerosis of neurogenic bladder research is limited. It is mainly for the treatment of the above control the symptoms, improve the function of urinary system, such as intermittent catheterization, anticholinergic drugs, botulinum toxin, capsaicin, surgery and so on, the treatment effect is not ideal, for many Hardening treatments such as immunosuppressants, immunoglobulin, plasma exchange and beta interferon effect cannot make people completely satisfied, so a new method of treatment for multiple sclerosis and neurogenic bladder. Recent studies have shown that gender differences will cause different incidence of autoimmune diseases women, in patients with multiple sclerosis patients is 2 times that of men, women in late pregnancy incidence of multiple sclerosis is reduced to 20% of the original, studies have shown that estrogen can prevent further deterioration of the disease, multiple sclerosis, promote nerve repair. There are also reported that estrogen receptor in bladder distribution, influence and estrogen on female bladder structure and function, the potential of estrogen in the treatment of multiple sclerosis of neurogenic bladder. Through this experiment, experimental autoimmune Encephalomyelitis (experimental autoimmune, encephalomyelitis, EAE) of the animal model to observe multiple sclerosis caused by bladder dysfunction, and estrogen treated animal model to observe the changes of the amount, animal model, preliminary verification of the possible therapeutic effect of estrogen on multiple sclerosis in neurogenic bladder. 1 experimental methods to establish EAE mice model: C57BL/6 female mice for 8-13 weeks, 200ug MOG35-55, and the volume of CFA content for complete Freund's adjuvant 10mg/ml mix, completely emulsion, subcutaneous injection points, in a mouse model of EAE 48 hours 400ng intraperitoneal injection of pertussis toxin induced immunity after immunization. Immunization day and since that day, observe and record the changes of body weight of rats, the function of clinical manifestations and neurological assessment.2 grouping and administration: 30 female C57BL6 mice were selected and randomly divided into 10 control group received only completely Fisheri Adjuvant and pertussis, 10 for EAE group, 10 for the estrogen treatment group, in complete immunization after treatment given daily doses of estrogen (50 mol/L) 100 L subcutaneous injection of.3 in the test specimen collection: 23 days, mice were anaesthetized with 10% chloral hydrate after removal of bladder tissue, weighing, preservation in liquid nitrogen, and then take the paraformaldehyde systemic perfusion, remove the mouse spinal cord tissue for the production of related indexes of paraffin section and.4 fast blue (LFB): Luxol staining and hematoxylin eosin (HE) staining of spinal cord inflammatory cell infiltration and demyelination situation, with the expression of estrogen receptor protein to detect the change of bladder Western-Blot. Results: 1 EAE mice model group clinical nerve function score, body weight, and urinary system function MS has typical clinical manifestations, HE staining showed obvious correlation of inflammatory cell infiltration showed.LFB staining An obvious demyelination of spinal.2 in multiple sclerosis of neurogenic bladder and bladder tissue in mice, compared with estrogen receptor mice significantly decreased in bladder control, estrogen treatment, estrogen receptor expression slightly upregulated the expression of.3 on EAE mice were given estrogen after treatment, mice reduced the incidence, severity of symptoms is not special EAE mice treated by light, the average neurological function score decreased. Conclusion 1 EAE neurological clinical symptoms score and multiple sclerosis of neurogenic bladder symptoms of.2 is closely related to estrogen and estrogen receptor in multiple sclerosis of neurogenic bladder diseases may play an important.3 in estrogen treatment on EAE model mice, the incidence and severity of symptoms compared with the untreated model group, estrogen treatment significantly decreased, suggesting that estrogen causes neurogenic bladder in multiple sclerosis Cysteine has an obvious improvement.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R744.51;R694.5

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