束缚应激对大鼠内脏感觉影响的研究

发布时间：2018-05-01 20:09

本文选题：急性束缚应激 + 慢性束缚应激　；参考：《华中科技大学》2006年博士论文

【摘要】： 第一部分急性和慢性束缚应激对大鼠内脏敏感性和神经内分泌的影响目的：了解急性和慢性束缚应激对大鼠内脏敏感性以及神经内分泌反应的影响特点和持续时间。方法：成年SD大鼠随机分为对照组(没有束缚应激)，急性部分束缚应激组(单次2h的束缚应激)和慢性部分束缚应激组(连续束缚应激3天，每天2h)，，通过腹壁回撤反射(AWR)评分评估应激前后大鼠内脏对结直肠扩张(CRD)的敏感性；并通过放免法检测应激前后不同时间点大鼠血浆促肾上腺皮质激素(ACTH)以及皮质酮(CORT)的水平。结果：(1)在扩张压力20，40mmHg时，急、慢性应激组的AWR评分均显著高于基础水平(0d)(P＜0.05)，但在应激后第7天(7d)AWR评分显著下降(P＜0.05)。(2)急性应激组血浆ACTH和CORT水平显著高于正常对照组(P＜0.05)，但是应激7日后，激素浓度下降至基础水平。(3)慢性应激组血浆ACTH和CORT水平在应激组也显著高于对照组，且激素水平在束缚7日后仍然维持在高水平(P＜0.05)。结论：急性和慢性束缚应激都可以导致大鼠内脏敏感性增高，但作用可能是短暂的。急性束缚应激短暂显著增加血浆ACTH和CORT的水平，而慢性束缚应激可能长期增加激素水平。神经内分泌激素的改变可以部分解释束缚应激导致内脏高敏性的原因。第二部分急性和慢性束缚应激对外周5-羟色胺能系统的影响目的：研究急性和慢性束缚应激对大鼠结肠5-HT合成的限速酶、5-HT转运体、5-HT1A和5-HT4受体的影响。方法：成年SD大鼠随机分为对照组(没有接受部分束缚应激(PRS))，急性部分束缚应激组(A-PRS)，急性部分束缚应激后恢复7日组(AR-PRS)，慢性部分束缚应激组(C-PRS)，慢性束缚应激后恢复7日组(CR-PRS)。采用RT-PCR半定量方法分别检测以上各组大鼠近端以及远端结肠组织中的色氨酸羟化酶(TpH)，5-HT转运体(5-HTT)，5-HT1A以及5-HT4受体的mRNA表达。结果：(1)与对照组相比，C-PRS组以及CR-PRS组大鼠的TpH以及5-HTT mRNA显著降低(C-PRS：0.69±0.13 vs.1.23±0.36 and 0.69±0.18 vs.1.20±0.35，P＜0.05；CR-PRS：0.70±0.12 vs.1.23±0.36 and 0.56±0.15 vs.1.20±0.35，P＜0.05)(2)急性束缚应激组和急性束缚应激后恢复7日组的TpH以及5-HTT的mRNA与正常组相比无显著变化(P＞0.05)。(2)5-HT1A和5-HT4受体的mRNA在A-PRS和C-PRS组均显著高于对照组(P＜0.05)。并且在恢复7日后，5-HT1A和5-HT4受体的mRNA仍维持在较高水平(P＜0.05)。结论：慢性束缚应激抑制肠道5-HT合成、代谢的分子，而急性束缚应激无显著影响；急性和慢性束缚应激均可以显著增加肠道5-HT受体的表达。急、慢性束缚应激对胃肠道功能失调的作用有不同机制。第三部分替加色罗对急性束缚应激导致内脏敏感性和脊髓c-fos表达的影响目的：研究替加色罗对急性束缚应激导致的内脏高敏感性和对脊髓相应节段c-fos表达的影响。方法：本实验分为两个部分。第一部分：雄性SD大鼠被随机分为正常对照组(假性束缚应激组)，束缚应激后腹腔注射替加色罗0.3mg／kg、0.6mg／kg、1.2mg／kg组，或束缚应激后腹腔注射溶酶0.3ml／kg、0.6ml／kg、1.2ml／kg对照组。通过观察结直肠扩张时(20，40，60，80mmHg)腹壁回撤反射评分，评估大鼠的内脏敏感性。第二部分：雄性SD大鼠随机分为束缚应激组，束缚应激后注射替加色罗1.2mg／kg组或溶媒1.2ml／kg组，以及正常对照组(假性束缚应激组)，每组大鼠均接受重复的结直肠扩张(60mmHg)2h。实验结束后取大鼠胸-腰段脊髓(T13，L1)和腰-骶段脊髓(L6，S1)，通过免疫组化方法检测大鼠脊髓c-fos表达。结果：(1)各剂量溶酶对照应激组的AWR评分在均显著高于正常对照组(P＜0.05)；替加色罗应激组AWR评分在各个剂量均显著低于相应剂量的溶酶应激对照组，并且呈量-效依赖性改变(P＜0.05)。(2)与正常对照组相比，束缚应激组或溶媒组可以显著增加结直肠扩张后胸腰段、以及腰骶段脊髓背角或侧柱的c-fos表达(P＜0.05)，注射替加色罗后会显著降低结直肠扩张后胸腰段、以及腰骶段脊髓背角或侧柱的c-fos表达(P＜0.05)。结论：替加色罗可以量-效依赖性降低急性应激导致的内脏感觉过敏；并且可以显著降低应激后结直肠扩张导致的胸腰段、以及腰骶段脊髓内脏输入区域的c-fos表达。
[Abstract]:Part one the effects of acute and chronic restraint stress on visceral sensitivity and neuroendocrine in rats
Objective: to understand the effects of acute and chronic restraint stress on the visceral sensitivity and the duration of neuroendocrine response in rats. Methods: adult SD rats were randomly divided into control group (without restraint stress), acute partial binding stress group (single 2H restraint stress) and chronic partial restraint stress group (3 days of continuous restraint stress, each). 2H), the abdominal wall retracement reflex (AWR) score was used to evaluate the sensitivity of the viscera to colorectal dilatation (CRD) before and after stress, and the level of plasma adrenocorticotropin (ACTH) and corticosterone (CORT) in rats at different time points before and after stress was detected by radioimmunoassay. Results: (1) AWR in the dilated pressure 20,40mmHg, acute, chronic stress group AWR The scores were significantly higher than the basic level (0d < 0.05) (P < 0.05), but the score of AWR was significantly decreased (P < 0.05) at seventh days after stress (P < 0.05). (2) the level of plasma ACTH and CORT in the acute stress group was significantly higher than that in the normal control group (P < 0.05), but the hormone concentration decreased to the base level after 7 days of stress. (3) the plasma ACTH and CORT levels in the chronic stress group were also shown in the stress group. It was higher than the control group, and the level of hormone remained at a high level after 7 days (P < 0.05). Conclusion: both acute and chronic restraint stress can lead to increased visceral sensitivity in rats, but the effect may be transient. Acute restraint stress temporarily increases the level of plasma ACTH and CORT, and chronic restraint stress may increase for a long time. The change of neuroendocrine hormone can partly explain the cause of visceral hypersensitivity induced by restraint stress.
The second part is the effect of acute and chronic restraint stress on peripheral 5- serotonin system.
Objective: To study the effect of acute and chronic restraint stress on the speed limit enzyme, 5-HT transporter, 5-HT1A and 5-HT4 receptor in the colon 5-HT synthesis in rats. Methods: adult SD rats were randomly divided into control group (without partial binding stress (PRS)), acute partial binding stress group (A-PRS), 7 days after acute partial restraint stress (AR-PRS), chronic part Shackle stress group (C-PRS) and 7 days after chronic restraint stress (CR-PRS). RT-PCR semi quantitative method was used to detect tryptophan hydroxylase (TpH) in proximal and distal colon tissues, 5-HT transporter (5-HTT), 5-HT1A, and mRNA expression of 5-HT4 receptor. Results: (1) compared with control group, C-PRS group and CR-PRS group The TpH and 5-HTT mRNA in rats were significantly decreased (C-PRS:0.69 + 0.13 vs.1.23 + 0.36 and 0.69 + 0.18 vs.1.20 + 0.35, P < 0.05, CR-PRS:0.70 + 0.12 vs.1.23 + 0.36 and 0.56 + 0.15 vs.1.20 0.35, 0.05) The changes (P > 0.05). (2) the mRNA of 5-HT1A and 5-HT4 receptors in A-PRS and C-PRS groups were significantly higher than those in the control group (P < 0.05). And the mRNA of 5-HT1A and 5-HT4 receptors remained at a higher level after 7 days (P < 0.05). Conclusion: chronic restraint stress inhibits intestinal 5-HT synthesis, metabolic molecules, and acute restraint stress has no significant effect; acute restraint stress has no significant influence; And chronic restraint stress can significantly increase the expression of 5-HT receptor in the intestinal tract. Acute, chronic restraint stress has different mechanisms on gastrointestinal dysfunction.
The third part is the effect of tegacrome on visceral sensitivity and c-fos expression in spinal cord after acute restraint stress.
Objective: To study the effect of Tegal on the sensibility of visceral Gao Min and the expression of c-fos in the corresponding segments of the spinal cord caused by acute restraint stress. Methods: this experiment was divided into two parts. The first part: male SD rats were randomly divided into normal control group (pseudorestraint stress group), Tegal 0.3mg / kg, 0.6mg / k after binding and intraperitoneal injection. G, 1.2mg / kg group, or intraperitoneal injection of enzyme 0.3ml / kg, 0.6ml / kg, 1.2ml / kg control group after restraint stress. By observing the reflex score of abdominal wall retracement in colorectal dilatation (20,40,60,80mmHg), the visceral sensitivity of rats was evaluated. The second part: male SD rats were randomly divided into bundle binding stress group, and Tegal stress after restraint stress. Group or solvent 1.2ml / kg group, and normal control group (pseudorestraint stress group), rats in each group received repeated colorectal dilatation (60mmHg) 2h. experiment to take the thoracic and lumbar spinal cord (T13, L1) and lumbar sacral spinal cord (L6, S1) after the end of the 2h. experiment. The results of immunohistochemistry were used to detect the expression of c-fos in the spinal cord of rats. Results: (1) each dose of enzyme control stress group The AWR score was significantly higher than that in the normal control group (P < 0.05), and the AWR score in the tegroline stress group was significantly lower than that in the corresponding enzyme stress control group, and the dose effect dependence was changed (P < 0.05). (2) compared with the normal control group, the restraint stress group or the solvent group could significantly increase the thoracic and lumbar segments after the colorectal distention, C-fos expression in the dorsal horn or lateral column of the lumbosacral spinal cord (P < 0.05), after injection of tegroroxo, the c-fos expression in the thoracic and lumbar segments after colorectal dilatation, as well as the c-fos expression of the dorsal horn or lateral column of the lumbosacral segment (P < 0.05). Conclusion: teanolo can reduce the visceral hypersensitivity caused by acute stress and can be significantly reduced. C-fos expression in the thoracolumbar spine and the visceral input area of the lumbosacral spinal cord after low stress.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R363

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