大鼠前列腺组织P物质与慢性前列腺炎的相关性研究
发布时间:2018-04-27 18:36
本文选题:慢性前列腺炎 + 神经内分泌细胞 ; 参考:《安徽医科大学》2014年硕士论文
【摘要】:目的通过对神经内分泌细胞分泌物P物质的研究来探讨大鼠神经内分泌细胞与慢性前列腺炎之间的相关性。 方法选取40只2月龄的SPF级健康雄性Sprague-Dawley(SD)大鼠作为实验对象,天平秤重,体重均在220-275g之间,,然后按体重排序编号,完全随机分为两组:模型组和对照组,每组20只,均在无菌环境中常规饲养。其中模型组SD大鼠经10%的水合氯醛麻醉后,在无菌条件下经腹行去势手术,手术当天及术后连续2天给予青霉素5万U/(kg·d)肌肉注射,一周后观察伤口无感染,愈合良好,开始背部皮下注射17β-雌二醇0.25mg/kg,连续注射30天;对照组SD大鼠未行去势手术,而在同一时间于背部皮下连续注射0.9%生理盐水一月。在最后一次注射17β-雌二醇和生理盐水的24小时以内,及时在无菌条件下运用心脏穿刺采血法收集5ml全血,然后迅速取出前列腺组织,放置于D-Hanks液中,再仔细剔除前列腺包膜及周围脂肪、血管等组织,以获得较为纯净的前列腺组织,最后用4%的多聚甲醛固定前列腺组织,并放入-80℃冰箱中备用。其中一部分前列腺组织经过固定、石蜡包埋、脱水、透明、浸蜡和切片后,行HE染色,光镜观察,发现模型组前列腺间质充血水肿,腺腔细胞不均匀增生,腺泡腔内和周围见大量炎症细胞浸润;而对照组SD大鼠的前列腺间质无水肿,无炎症细胞浸润,腺泡上皮排列整齐,无增生现象。这表明SD大鼠慢性前列腺炎模型建造成功;而另一部分前列腺组织则经过上述的一系列处理后用于免疫组织化学实验;运用心脏穿刺法收集的全血经过离心后,取其上清液移入无菌瓶内,然后采用双抗体夹心ELISA法测定两组SD大鼠前列腺组织中P物质表达的差异,并观察此差异在统计学意义上是否具有显著性。 结果免疫组化结果:模型组中P物质染色阳性细胞数较对照组显著增加且大都围绕在前列腺管周围,而对照组散在存在少量的P物质染色阳性细胞,其中模型组及对照组中P物质的阳性表达率分别为85%(其中弱阳性3例,阳性8例,强阳性6例)和20%(其中弱阳性2例,阳性2例),进行组间比较,发现差异具有统计学意义(P0.01);ELISA结果:模型组中P物质的含量为(113.5±17.5)ng/ml;对照组中P物质的含量为(98.0±18.1)ng/ml,组间比较,差异在统计学上具有显著性(t=2.742,P=0.009)。结论本实验结果显示模型组SD大鼠血清及前列腺组织中的P物质表达明显高于对照组,这表明神经内分泌细胞与慢性前列腺炎之间存在着密切的关系,其中的作用机制可能为神经内分泌细胞通过其活性分泌产物P物质(SP)、嗜铬蛋白A(CgA)、神经元特异性烯醇化酶(NSE)及神经生长因子(NGF)等作用于前列腺上皮细胞及平滑肌细胞并影响其功能,从而引起一系列慢性前列腺炎样症状。因此神经内分泌细胞对前列腺的生长、分化和分泌功能等起着一定的调节作用,它可能参与慢性前列腺炎的病因和发病过程。
[Abstract]:Objective to investigate the relationship between neuroendocrine cells and chronic prostatitis by studying substance P from neuroendocrine cells. Methods Forty healthy SPF male Sprague-Dawley SD rats aged 2 months were selected as experimental subjects, weighing between 220 and 275 g, and then randomly divided into two groups: model group and control group, 20 rats in each group, and 20 rats in each group were randomly divided into two groups: model group and control group, each group was divided into two groups: model group (n = 20), control group (n = 20) and control group (n = 20). All of them were fed in aseptic environment. SD rats in the model group were anesthetized by 10% chloral hydrate, castrated by abdominal castration under aseptic conditions. Penicillin was injected intramuscularly on the day of operation and 2 days after operation. No infection was observed after one week, and the wound healed well. 17 尾 -estradiol 0.25 mg / kg was injected subcutaneously into the back for 30 days, while the control group did not undergo ovariectomized, but at the same time, 0.9% saline was injected subcutaneously on the back for one month. Within 24 hours of the last injection of 17 尾 -estradiol and normal saline, the whole blood of 5ml was collected by cardiac puncture in time under aseptic condition, and then the prostate tissue was quickly removed and placed in D-Hanks fluid. Then carefully remove the prostate capsule, surrounding fat, blood vessels and other tissues in order to obtain more pure prostate tissue, and finally fixed the prostate tissue with 4% paraformaldehyde, and put it in the refrigerator at -80 鈩
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