本文選題：老年　切入點(diǎn)：脊髓腰骶段　出處：《河北醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：脊髓腰骶段對調(diào)控機(jī)體盆底肌、大腸和生殖器官等起著重要作用。在機(jī)體衰老的過程中，脊髓腰骶段呈現(xiàn)出與年齡相關(guān)的衰老性變化，同時還會表現(xiàn)出腸、膀胱和生殖器官等盆腔臟器的器質(zhì)性與功能性改變。一氧化氮（Nitric oxide，NO）是一種在外周和中樞神經(jīng)系統(tǒng)中起著重要信號轉(zhuǎn)導(dǎo)作用的神經(jīng)信息物質(zhì)。研究顯示盆腔臟器的功能異�？赡芘c老年脊髓腰骶段的NO表達(dá)變化有關(guān)。有研究報(bào)道在衰老的過程中脊髓腰骶段出現(xiàn)年齡相關(guān)的還原型尼克酰胺腺嘌呤二核苷酸磷酸黃遞酶陽性體（Age-relatednicotinamide adenine dinucleotide phosphate-diaphorase positive bodies，ANB）的表達(dá)。 隨著機(jī)體的衰老，老年男性經(jīng)常出現(xiàn)排尿功能的障礙，這可能與排尿器官或參與排尿反射的神經(jīng)調(diào)節(jié)的衰老性改變有關(guān)。參與機(jī)體排尿反射神經(jīng)調(diào)節(jié)的低位中樞位于脊髓的腰骶段，脊髓腰骶段出現(xiàn)的ANB可能影響了機(jī)體的排尿反射的神經(jīng)調(diào)節(jié)功能。在衰老的過程中，體內(nèi)雄激素-睪酮（Testosterone，T）逐漸減少，ANB的表達(dá)變化是否與體內(nèi)T的減少有關(guān)目前尚不清楚。脊髓腰骶段5-羥色胺（5-hydroxytryptamine，5-HT）能和多巴胺（Dopamine，DA）能的信息傳遞在控制排尿的反射中起著重要的作用，除NO特性之外，ANB是否還具有其它神經(jīng)化學(xué)成分，如5-HT\DA值得進(jìn)一步研究。 目的：觀察老年大鼠脊髓腰骶段ANB的表達(dá)以及丙酸睪酮（Testosterone propionate，TP）對其表達(dá)的影響，探討ANB是否與5-TH、TH共存。 方法：選取健康雄性Wistar大鼠，將實(shí)驗(yàn)動物分成6月齡組（6Mon）、21月齡（21Mon）組和24月齡（24Mon）組。 將21Mon組和24Mon組分別采取皮下和滴鼻兩種給藥方式，即21月齡皮下給藥（21Mon-sc.TP）、21月齡滴鼻給藥（21Mon-int.TP）、24月齡皮下給藥（24Mon-sc.TP）和24月齡滴鼻給藥（24Mon-int.TP）組。分別設(shè)定各自給藥方式的對照（21Mon-sc.con、21Mon-int.con、24Mon-sc.con、24Mon-int.con）組。每組各5只。皮下給藥組通過背部皮下注射睪酮制劑-TP；滴鼻給藥組通過鼻腔給予TP。給藥劑量為2.0mg/kg.d，于5:00~6:00PM期間給藥，每天1次。對照組分別按上述方式給予芝麻油作為安慰劑。連續(xù)給藥3個月（即21Mon組從18個月開始給藥，24Mon組從21個月開始給藥）。 大鼠以4%多聚甲醛經(jīng)心灌注固定，取脊髓腰骶段30%蔗糖保護(hù)后，行冰凍切片。相應(yīng)切片以NADPH-d組織化學(xué)染色法顯示ANB的表達(dá)、以NADPH-d組織化學(xué)復(fù)合5-HT或TH免疫組織化學(xué)雙標(biāo)記法觀察ANB與5-HT、TH是否共存。 結(jié)果： 1. NADPH-d組織化學(xué)染色的結(jié)果： （1）切片的一般觀察：大鼠脊髓腰骶段的NOS陽性神經(jīng)元被染成藍(lán)色，細(xì)胞呈高爾基樣。NOS陽性神經(jīng)元胞體以中等大小為主，深染或淡染，呈梭形、三角形或圓形；有2-4個突起，胞核呈空泡狀不著色。在21Mon組和24Mon組中，位于脊髓腰骶段的SDH、LT、ILN、DCN、CC等部位可見ANB的出現(xiàn)，呈現(xiàn)球形或不規(guī)則形，深染邊緣銳利且密度高。 （2）ANB的數(shù)量變化：經(jīng)觀察，在6Mon組未見ANB的出現(xiàn)；21Mon和24Mon組在SDH、LT、ILN、DCN、CC處均可見ANB，并且在24Mon組的數(shù)目多于21Mon組。經(jīng)統(tǒng)計(jì)，21Mon-sc.con組與21Mon-int.con組無明顯區(qū)別（P0.05）；24Mon-sc.con組和24Mon-int.con組無明顯區(qū)別（P0.05）。與21Mon-int.con組相比，，ANB在24Mon-int.con組的SDH、LT、ILN、DCN和CC數(shù)目分別增長了40%（P0.05）、65%（P0.05）、69%（P0.05）、88%（P0.05）和39%（P0.05），有統(tǒng)計(jì)學(xué)意義。 （3）丙酸睪酮對ANB的影響：①皮下給予TP對ANB的影響：與21Mon-sc.con組相比，ANB在21Mon-sc.TP組的SDH、LT、ILN、DCN和CC的數(shù)目分別減少了40%（P0.05）、34%（P0.05）、40%（P0.05）、60%（P0.05）和82%（P0.05），有統(tǒng)計(jì)學(xué)意義。與24Mon-sc.con組相比，在24Mon-sc.TP組的SDH、LT、ILN和DCN的數(shù)目分別減少了5%、13%、3%和6%，在CC增加了8%。除后者外，所觀察部位存在ANB數(shù)目下降的趨勢。②鼻腔給予TP對ANB的影響：與21Mon-int.con組相比，ANB在21Mon-int.TP組的SDH、LT、ILN、DCN和CC的數(shù)目分別減少了20%（P0.05）、22%（P0.05）、32%（P0.05）、38%（P0.05）和44%（P0.05），有統(tǒng)計(jì)學(xué)意義。與24Mon-int.con組相比，ANB在24Mon-int.TP組的ILN、DCN和CC分別減少6%、10%和10%。部分核團(tuán)ANB數(shù)目有下降的趨勢，但沒有統(tǒng)計(jì)學(xué)意義。（4）21Mon組皮下和鼻腔給予TP對ANB影響的區(qū)別：21Mon組大鼠滴鼻給藥和皮下給藥后都可以使ANB數(shù)目下降，兩種給藥途徑對ANB數(shù)量的影響無顯著區(qū)別。 2. NADPH-d組織化學(xué)復(fù)合5-HT/TH免疫組織化學(xué)：ANB未見與5-HT或TH免疫反應(yīng)產(chǎn)物的共存。 小結(jié)：1.脊髓腰骶段ANB的出現(xiàn)與衰老有關(guān)，隨著機(jī)體的老化ANB的數(shù)目逐漸增多。2.脊髓腰骶段ANB的出現(xiàn)可能與體內(nèi)衰老過程中睪酮的減少存在一定的關(guān)系，補(bǔ)充丙酸睪酮可以減少衰老過程中ANB的出現(xiàn)。3.鼻腔給予丙酸睪酮與皮下給予丙酸睪酮對ANB的影響效果相同。4. ANB與5-HT和DA能神經(jīng)元未見共存。
[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錛

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