论文标题:糖尿病大鼠脑干nNOS免疫阳性神经元的变化
Changes of Neuronal Nitric Oxide Synthase Immunopositive Neurons in Brain Stem of Diabetic Rats
论文作者 林宏生
论文导师 沈伟哉,论文学位 硕士,论文专业 人体解剖学
论文单位 暨南大学,点击次数 89,论文页数 50页File Size2723k
2003-04-01论文网 http://www.lw23.com/lunwen_970545162/ 糖尿病,脑干,被盖背外侧核,蓝斑下核,中央灰质背侧核,神经元型一氧化氮合酶,一氧化氮,大鼠
diabetes mellitus;brain stem;laterodorsal tegmental nucleus,subcoeruleus nucleus;dorsal central gray nucleus;neuronal nitric oxide synthase;nitric oxide;rat
目的：观察糖尿病大鼠脑桥被盖背外侧核、蓝斑下核、中央灰质背侧核nNOS免疫阳性神经元的的变化，探讨NO对糖尿病及其中枢神经系统慢性病变的作用机制。 方法：SD大鼠36只，随机分为2组，(1)对照组；(2)糖尿病组：用链脲佐菌素诱导建立胰岛素依赖型糖尿病大鼠模型。建模后2、7、12周末处死、取材。ABC免疫细胞化学方法观察脑桥被盖背外侧核、蓝斑下核、中央灰质背侧核nNOS免疫阳性神经元的变化，并用体视学方法进行定量分析，同时观察血糖、血清NO含量。 结果：(1)糖尿病大鼠脑桥被盖背外侧核nNOS免疫阳性神经元的数量在2周、7周、12周均较对照组明显增多(P＜0.01)并基本保持稳定；其平均光密度(AOD)、体积分光密度(VIOD)在2周、7周、12周均较对照组明显升高(P＜0.01)。(2)糖尿病大鼠蓝斑下核nNOS免疫阳性神经元的数量在2周、7周、12周均较对照组明显增多(P＜0.01)；其AOD、VIOD虽然在7周、12周较2周略下降，但均明显高于对照组(P＜0.01)。(3)糖尿病大鼠中央灰质背侧核nNOS免疫阳性神经元在2周、7周、12周亦均较对照组明显增多(P＜0.01)；其AOD、VIOD在2周、7周、12周的变化与蓝斑下核相似，明显高于对照组(P＜0.01)。(4)糖尿病大鼠血清NO含量在糖尿病2周明显升高(P＜0.01)，在糖尿病7周、12周下降至正常水平。 结论：(1)糖尿病大鼠脑干内nNOS免疫阳性神经元与外周血清NO的变化存在不平行现象，可能是由于外周eNOS催化生成NO出现障碍所致。(2)糖尿病大鼠脑干nNOS免疫阳性神经元的数量以及nNOS免疫阳性物质的含量显著增多，提示增多的NO可能通过其对神经细胞的毒性作用造成脑组织内神经细胞结构和功能的破坏。(3)糖尿病大鼠脑桥被盖背外侧核、蓝斑下核nNOS免疫阳性神经元的数量以及nNOS免疫阳性物质含量的显著增多可能与糖尿病晚期睡眠障碍有关。(4)糖尿病大鼠中央灰质背侧核和蓝斑下核nNOS免疫阳性神经元的数量以及nNOS免疫阳性物质含量的显著增多可能与糖尿病晚期感觉过敏或感觉丧失有关。
Objective: To investigate the role of nitric oxide in diabetes mellitus and diabetic neuropathy in central nervous system by observing the changes of neuronal nitric oxide synthase (nNOS) immunopositive neurons in laterodorsal tegmental nucleus (LDT), subcoeruleus nucleus (SubC), dorsal central gray nucleus (DCG) and analyzing quantitatively with image analyzer in diabetic rats.Methods: 36 SD rats were divided randomly into control and diabetic groups. The diabetic group was first established STZ-induced diabetic rats, and then was sacrificed and the materials were drawn at the end of 2, 7 and 12 weeks. The number of nNOS immunopositive neurons in LDT, SubC and DCG was counted and analysed quantitatively by image analyzer. The changes of serum glucose and serum nitric oxide at every period were also analysed.Results: (1) The number of nNOS immunopositive neurons of LDT of the diabetic group in 2 weeks, 7 weeks and 12 weeks was significantly increased by comparison with that of the control group and kept relatively stable (P<0.01). The average optical density (AOD) and volum integrated optical density (VIOD) of the diabetic group in 2 weeks, 7 weeks and 12 weeks were more than that of the control group (P<0.01). (2) The number of nNOS immunopositive neurons of SubC of the diabetic group in 2 weeks, 7 weeks and 12 weeks was significantly increased (P<0.01), when compared with that of the control group. The AOD and VIOD of the diabetic group in course of the experiment were decreased slightly, however, significantly increased in comparison with that of the control group (P<0.01). (3) The number of nNOS immunopositive neurons of DCG of the diabetic group in 2 weeks, 7 weeks and 12 weeks was significantly increased when compared with that of the control group (P<0.01). The AOD and VIOD of diabetic group were significantly increased when performing an act of comparison with that of the control group (P<0.01). (4) The concentration of nitric oxide in serum of the diabetic group increased significantly in 2 weeks (P<0.01), then decreased to normal level from 7 weeks. Conclusions: (1) The number of nNOS immunopositive neurons and the content ofnNOS immunopositive substance in LDT, SubC and DCG were different from the content of nitric oxide in peripheral serum. It may be related to the lack of nitric oxide by catalysis of eNOS in peripheral serum. (2) The number of nNOS immunopositive neurons and the content of nNOS immunopositive substance in LDT, SubC and DCG were significantly increased. It suggested that the increased nitric oxide might be result in neurocyte toxic effect of nitric oxide that could destroy cellular architecture and function. (3) The number of nNOS immunopositive neurons and the content of nNOS immunopositive substance in LDT and SubC were significantly increased. It suggested that the increased nitric oxide might be related to sleep disorder that often occurred in diabetic late stage. (4) The number of nNOS immunopositive neurons and the content of nNOS immunopositive substance in DCG and SubC were significantly increased. It may be associated with sensory disturbance such as hyperesthesia and sensory deprivation in diabetic late stage.
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