BNP及NPR-A在慢性非细菌性前列腺炎大鼠模型L6-S1神经节中的表达及其意义

发布时间：2018-02-24 23:27

本文关键词： 慢性非细菌性前列腺炎完全弗氏佐剂 BNP NPR-A 疼痛　出处：《南昌大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：建立稳定的慢性非细菌性前列腺炎（CNP）SD大鼠模型。通过检测假手术对照组及CNP大鼠模型探讨脑钠尿肽（BNP）及脑钠尿肽受体(NPR-A)在慢性非细菌性前列腺炎(CNP)大鼠脊髓背角神经节L6-S1（DRG）中的表达情况，观察BNP及NPR-A的表达变化，以及CNP引起的慢性病理性疼痛与BNP、NPR-A之间的关系。方法： 1.本实验在南昌大学第一附属医院泌尿外科研究所内完成。选择清洁型SD大鼠100只（全部为雄性），适应性饲养1周后，随机分为5组分别为：对照组,向前列腺内注射生理盐水0.1ml;实验组：3d造模组、7d造模组、10d造模组，14d造模组，向前列腺内注射完全弗氏佐剂（CFA）0.1ml构建慢性非细菌性前列腺炎大鼠模型，以上每组均为20只。 2.根据不同的造模时间点分别提取对照组及各个实验组SD大鼠的脊髓背角L6-S1段神经节。 3.将其中各组10只的L6-S1神经节段组织中提取RNA，，进行逆转录成cDNA、再行实时荧光定量PCR，观察和计算在对照组及造模后各个时间点中BNP及NPR-A表达及其差异，研究BNP、NPR-A在慢性非细菌性前列腺炎中的表达、意义及其相关性。 4.应用免疫组化SP方法检测另外各组10只的L6-S1神经节中BNP、NPR-A中的表达及其差异，研究BNP、NPR-A在慢性非细菌性前列腺炎中的表达、意义及其相关性。结果： 1.荧光定量PCR所显示的结果是BNP在实验造模组的表达量比较对照组均为上调。3d造模组、7d造模组、10d造模组，14d造模组比较对照组分别是1.53±0.28倍，2.10±0.32倍，1.85±0.26倍，1.77±0.35倍。荧光定量PCR所显示的结果是NPR-A在实验造模组的表达量比较对照组均为上调。3d造模组、7d造模组、10d造模组，14d造模组比较对照组分别是1.47±0.21倍，2.05±0.26倍，1.98±0.31倍，1.67±0.23倍。其中7d造模组、10d造模组和14d造模组与对照组比较差异，有统计学意义（P＜0.05）。 2.免疫组化方法所显示的结果是BNP及NPR-A在神经节中的神经纤维上着色，染色呈红色，并且在实验造模组的染色强度高于对照组。BNP及NPR-A的平均光密度值（OD值）在实验造模组的表达量比较对照组均为上调。其中7d、10d和14d造模组与对照组比较差异具有统计学意义（*P＜0.05）。结论：利用完全弗氏佐剂（CFA）前列腺注射法建立稳定的CNP大鼠模型。BNP及NPR-A在SD大鼠L6-S1神经节中均有表达，而且造模后形成前列腺炎症后其表达均上调。在CNP导致的疼痛机制中，BNP及NPR-A有可能参与其中，以及BNP/NPR-A信号通路在CNP中可能被激活。
[Abstract]:Objective:. A stable rat model of chronic nonbacterial prostatitis was established. To investigate the effect of brain natriuretic peptide (BNP) and brain natriuretic peptide receptor (NPR-A) on the spinal cord of rats with chronic nonbacterial prostatitis (CNP) by detecting the sham operation control group and the CNP rat model. The expression of L6-S1 DRG in hornular ganglion, The expression of BNP and NPR-A and the relationship between chronic pathological pain induced by CNP and NPR-A were observed. Methods:. 1. The experiment was carried out in the Institute of Urology, the first affiliated Hospital of Nanchang University. 100 clean SD rats (all male rats) were randomly divided into 5 groups: control group after one week of adaptive feeding. The rats in the experimental group were injected with normal saline 0.1 ml into the prostate, the experimental group was divided into two groups: the control group (n = 7), the control group (n = 10), the control group (n = 14), and the control group (n = 20) were injected with complete Freund's adjuvant CFAA (0.1 ml) into the prostate to establish the model of chronic non-bacterial prostatitis (n = 20). 2. L6-S1 segment ganglion of spinal dorsal horn was extracted from SD rats of control group and experimental group according to different time points. 3. RNAs were extracted from L6-S1 nerve segments of 10 rats in each group, and then reverse transcripted into cDNAs. The expression and difference of BNP and NPR-A were observed and calculated in the control group and at each time point after modeling. To study the expression, significance and correlation of BNPN NPR-A in chronic nonbacterial prostatitis. 4. Immunohistochemical SP method was used to detect the expression and difference of BNPNPR-A in L6-S1 ganglion, and to study the expression and significance of BNPNPR-A in chronic non-bacterial prostatitis. Results:. 1. The results of fluorescence quantitative PCR showed that the expression of BNP in the experimental model group was higher than that in the control group. The expression of BNP in the control group was 1.53 卤0.28 times 2.10 卤0.32 times 1.85 卤0.26 times 1.77 卤0.35 times, respectively. The results of PCR showed that the expression of NPR-A in the experimental model group was higher than that in the control group on the 7th day and the 7th day in the control group. The comparison of the control group was 1.47 卤0.21 times and 2.05 卤0.26 times and 1.98 卤0.31 times and 1.67 卤0.23 times respectively, and the control group was 1.47 卤0.21 times and 1.98 卤0.31 times and 1.67 卤0.23 times respectively. The difference between the control group and the control group was observed. There was statistical significance (P < 0.05). 2. The results of immunohistochemistry showed that BNP and NPR-A were stained on the nerve fibers in the ganglion, and the staining was red. The staining intensity of the experimental model group was higher than that of the control group and the average optical density value (OD value of NPR-A) in the experimental model group, the expression level of the experimental model group was higher than that of the control group, and there was significant difference between the model group and the control group on the 10th and 14th days after 7 days (P < 0.05). Conclusion:. The stable CNP rat model. BNP and NPR-A were expressed in the L6-S1 ganglion of SD rats by using complete Freund's adjuvant CFA-prostatic injection. The expression of BNP and NPR-A may be involved in the pain mechanism induced by CNP, and the BNP/NPR-A signaling pathway may be activated in CNP.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R697.33

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