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老年大鼠脊髓腰骶段ANB表达及相关特性

发布时间:2018-03-03 14:18

  本文选题:老年 切入点:脊髓腰骶段 出处:《河北医科大学》2012年硕士论文 论文类型:学位论文


【摘要】:脊髓腰骶段对调控机体盆底肌、大肠和生殖器官等起着重要作用。在机体衰老的过程中,脊髓腰骶段呈现出与年龄相关的衰老性变化,同时还会表现出肠、膀胱和生殖器官等盆腔脏器的器质性与功能性改变。一氧化氮(Nitric oxide,NO)是一种在外周和中枢神经系统中起着重要信号转导作用的神经信息物质。研究显示盆腔脏器的功能异常可能与老年脊髓腰骶段的NO表达变化有关。有研究报道在衰老的过程中脊髓腰骶段出现年龄相关的还原型尼克酰胺腺嘌呤二核苷酸磷酸黄递酶阳性体(Age-relatednicotinamide adenine dinucleotide phosphate-diaphorase positive bodies,ANB)的表达。 随着机体的衰老,老年男性经常出现排尿功能的障碍,这可能与排尿器官或参与排尿反射的神经调节的衰老性改变有关。参与机体排尿反射神经调节的低位中枢位于脊髓的腰骶段,脊髓腰骶段出现的ANB可能影响了机体的排尿反射的神经调节功能。在衰老的过程中,体内雄激素-睾酮(Testosterone,T)逐渐减少,ANB的表达变化是否与体内T的减少有关目前尚不清楚。脊髓腰骶段5-羟色胺(5-hydroxytryptamine,5-HT)能和多巴胺(Dopamine,DA)能的信息传递在控制排尿的反射中起着重要的作用,除NO特性之外,ANB是否还具有其它神经化学成分,如5-HT\DA值得进一步研究。 目的:观察老年大鼠脊髓腰骶段ANB的表达以及丙酸睾酮(Testosterone propionate,TP)对其表达的影响,探讨ANB是否与5-TH、TH共存。 方法:选取健康雄性Wistar大鼠,将实验动物分成6月龄组(6Mon)、21月龄(21Mon)组和24月龄(24Mon)组。 将21Mon组和24Mon组分别采取皮下和滴鼻两种给药方式,即21月龄皮下给药(21Mon-sc.TP)、21月龄滴鼻给药(21Mon-int.TP)、24月龄皮下给药(24Mon-sc.TP)和24月龄滴鼻给药(24Mon-int.TP)组。分别设定各自给药方式的对照(21Mon-sc.con、21Mon-int.con、24Mon-sc.con、24Mon-int.con)组。每组各5只。皮下给药组通过背部皮下注射睾酮制剂-TP;滴鼻给药组通过鼻腔给予TP。给药剂量为2.0mg/kg.d,于5:00~6:00PM期间给药,每天1次。对照组分别按上述方式给予芝麻油作为安慰剂。连续给药3个月(即21Mon组从18个月开始给药,24Mon组从21个月开始给药)。 大鼠以4%多聚甲醛经心灌注固定,取脊髓腰骶段30%蔗糖保护后,行冰冻切片。相应切片以NADPH-d组织化学染色法显示ANB的表达、以NADPH-d组织化学复合5-HT或TH免疫组织化学双标记法观察ANB与5-HT、TH是否共存。 结果: 1. NADPH-d组织化学染色的结果: (1)切片的一般观察:大鼠脊髓腰骶段的NOS阳性神经元被染成蓝色,细胞呈高尔基样。NOS阳性神经元胞体以中等大小为主,深染或淡染,呈梭形、三角形或圆形;有2-4个突起,胞核呈空泡状不着色。在21Mon组和24Mon组中,位于脊髓腰骶段的SDH、LT、ILN、DCN、CC等部位可见ANB的出现,呈现球形或不规则形,深染边缘锐利且密度高。 (2)ANB的数量变化:经观察,在6Mon组未见ANB的出现;21Mon和24Mon组在SDH、LT、ILN、DCN、CC处均可见ANB,并且在24Mon组的数目多于21Mon组。经统计,21Mon-sc.con组与21Mon-int.con组无明显区别(P0.05);24Mon-sc.con组和24Mon-int.con组无明显区别(P0.05)。与21Mon-int.con组相比,,ANB在24Mon-int.con组的SDH、LT、ILN、DCN和CC数目分别增长了40%(P0.05)、65%(P0.05)、69%(P0.05)、88%(P0.05)和39%(P0.05),有统计学意义。 (3)丙酸睾酮对ANB的影响:①皮下给予TP对ANB的影响:与21Mon-sc.con组相比,ANB在21Mon-sc.TP组的SDH、LT、ILN、DCN和CC的数目分别减少了40%(P0.05)、34%(P0.05)、40%(P0.05)、60%(P0.05)和82%(P0.05),有统计学意义。与24Mon-sc.con组相比,在24Mon-sc.TP组的SDH、LT、ILN和DCN的数目分别减少了5%、13%、3%和6%,在CC增加了8%。除后者外,所观察部位存在ANB数目下降的趋势。②鼻腔给予TP对ANB的影响:与21Mon-int.con组相比,ANB在21Mon-int.TP组的SDH、LT、ILN、DCN和CC的数目分别减少了20%(P0.05)、22%(P0.05)、32%(P0.05)、38%(P0.05)和44%(P0.05),有统计学意义。与24Mon-int.con组相比,ANB在24Mon-int.TP组的ILN、DCN和CC分别减少6%、10%和10%。部分核团ANB数目有下降的趋势,但没有统计学意义。(4)21Mon组皮下和鼻腔给予TP对ANB影响的区别:21Mon组大鼠滴鼻给药和皮下给药后都可以使ANB数目下降,两种给药途径对ANB数量的影响无显著区别。 2. NADPH-d组织化学复合5-HT/TH免疫组织化学:ANB未见与5-HT或TH免疫反应产物的共存。 小结:1.脊髓腰骶段ANB的出现与衰老有关,随着机体的老化ANB的数目逐渐增多。2.脊髓腰骶段ANB的出现可能与体内衰老过程中睾酮的减少存在一定的关系,补充丙酸睾酮可以减少衰老过程中ANB的出现。3.鼻腔给予丙酸睾酮与皮下给予丙酸睾酮对ANB的影响效果相同。4. ANB与5-HT和DA能神经元未见共存。
[Abstract]:The lumbosacral spinal cord on the regulation of pelvic floor muscle, intestine and reproductive organs plays an important role in the process of aging. In the lumbosacral spinal cord showed age-related changes associated with age, but also showed intestinal, organic and functional bladder and reproductive organs and other pelvic organs. The change of nitric oxide (Nitric oxide NO) is a kind of peripheral and central nervous system plays an important role in signal transduction of neural information material. Research shows that pelvic organ dysfunction can in elderly patients with lumbosacral spinal cord NO expression change. Studies have reported in the process of aging the lumbosacral spinal cord of age-related reduced nicotinamide adenine dinucleotide phosphate diaphorase positive body (Age-relatednicotinamide adenine dinucleotide phosphate-diaphorase positive bodies, ANB) expression.
With the aging of the body, often elderly male urination disorders, which may regulate aging and urinary organs or involved in the micturition reflex nerve changes. In the central regulation of low body micturition reflex nerve of lumbosacral spinal cord in the lumbosacral spinal cord of the ANB may affect the body's micturition reflex the nerve regulation function. In the process of aging in vivo androgen testosterone (Testosterone, T) decreased gradually, the expression of ANB and T in whether reduction is unclear. The lumbosacral spinal cord (5-hydroxytryptamine, 5-HT) 5- serotonin and dopamine (Dopamine, DA) to the information transmission in reflex control urination plays an important role in NO characteristics, ANB also has other neurochemical components, such as 5-HTDA deserves further study.
Objective: To observe the expression of ANB in the lumbosacral region of spinal cord in aged rats and the effect of Testosterone propionate (TP) on its expression, and to explore whether ANB coexists with 5-TH and TH.
Methods: healthy male Wistar rats were selected and the experimental animals were divided into 6 month old groups (6Mon), 21 month old (21Mon) group and 24 month old (24Mon) group.
The 21Mon group and 24Mon group respectively by subcutaneous and intranasal two medication, 21 month old subcutaneous administration (21Mon-sc.TP), 21 month old (21Mon-int.TP) administered intranasally administered subcutaneously (24Mon-sc.TP, 24 month old) and 24 month old (24Mon-int.TP) administered intranasally administered group. Each set the control respectively (21Mon-sc.con 21Mon-int.con, 24Mon-sc.con, 24Mon-int.con) group. Each group had 5 rats. Subcutaneous administration group by subcutaneous injection of testosterone preparations -TP; intranasal administration group by intranasal administration of TP. dose of 2.0mg/kg.d, 5:00~6:00PM during the administration, 1 times a day. The control group were given according to the above sesame oil as placebo. For 3 months (21Mon group administered 24Mon group from 18 months to 21 months from the beginning of administration).
The rats in 4% paraformaldehyde through heart perfusion fixation, the lumbosacral spinal cord of 30% sucrose protection, frozen sections. The corresponding sections in NADPH-d staining showed that the expression of ANB, NADPH-d or TH composite 5-HT histochemical immunohistochemical double labeling method was used to observe the ANB and 5-HT, TH is not coexist.
Result锛

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