摘要
背景:脑干孤束核是内脏痛信息向高位脑中枢传递的一个关键的中继站。形态学证据表明,大鼠孤束核中分布有大量的神经激肽B受体(neurokininBreceptors,NKR)。然而,孤束核中表达NKR的神经元是否参与内脏痛信息的传入与整合目前仍不清楚。目的:阐明孤束核中表达NKR的神经元在脑肠通路中传递内脏痛信息中的作用。设计:随机的对照实验性研究。地点和对象:12只SD大鼠购自第四军医大学实验动物中心。大鼠均为雄性,体质量220～250g,用随机数字表法分为实验组和对照组,每组各6只。干预:6只雄性大鼠吸入麻醉后,将2mL50g/L的乙酸经导管注入大鼠升结肠内,20s后再注入5mL生理盐水冲洗结肠。对照组大鼠则注射等体积的磷酸缓冲液(0.01mol/L)。24h后大鼠开胸经心脏灌流40g/L的甲醛。灌注毕后取出鼠脑修块,然后将含有孤束核的脑块在300g/L的蔗糖溶液中浸泡过夜,冰冻切片机切片。用抗FOS抗体和抗NKR抗体行免疫组化双重标记染色。免疫组化由第一作者完成,动物模型构建由第二作者完成,第三及第四作者负责免疫组化结果评估。主要观察指标:孤束核中NKR和FOS蛋白的共表达。结果:孤束核各个亚核中均有大量的NKR免疫阳性的神经元,但在内侧亚核和联合亚核中尤其浓密。阳性产物分布在细胞膜和树突上;胃肠道给予甲醛刺激后,表达FOS蛋?
BACKGROUND:The nucleus tractus solitarius(NTS) is one of the key relay stations of various visceral primary afferent information.Morphological evidence has shown that neurokinin B receptors(NKR) is densely distributed in the rat NTS.However,it still remains unclear whether the neurons expressing NKR in NTS are involved in the integration or modulation of visceral pain transmission. OBJECTIVE:To elucidate the possible involvement of NKR-positive neurons in NTS in the transmission or integration of visceral pain in the brain-gut pathway.DESIGN:Randomized controlled trial. SETTING and MATERIALS:Twelve male Sprague-Dawley rats weighing 220-250 g were randomized into experimental group(n=6) and control group(n=6) with table of random permutations.INTERVENTIONS:After the rats were anesthetized with methoxyflurane gas inhalation,a canule was quickly inserted into the ascending colon of the rats in the experimental group,through which 2 mL 50 g/L acetic acid was injected followed 20 s later by 5 mL 9 g/L saline to remove acetic acid from the colon.In the control group,2 mL 0.01 mol/L PBS were used.The animals were all intracardially perfused with 40 g/L paraformaldehyde 24 later,and the brain tissue blocks containing NTS were trimmed,immersed in 300 g/L sucrose solution overnight,and sliced on a freezing microtome.Double immunolabeling with mouse antisera against Fos and rabbit antisera against NKR was employed. MAIN OUTCOME MEASURES:Observations were made of the distributions of NKR and Fos proteins, and their co-expression in NTS.RESULTS:A great number of NKR-like immunoreactive(LI) neurons were identified in all the subnuclei of NTS,which were much more dense in the medial and commissural subnuclei,with the DAB-immunoreactive products confined on the cell membranes and the dendrites.Neurons expressing Fos protein following formalin irritation were predominantly distributed in the medial and commissural subnuclei of NTS.The characteristic neurons co-expressing NKR and Fos protein possessed black nuclei and brownish cel
出处
《中国临床康复》
CSCD
2004年第7期1358-1360,T004,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(39970377)~~
