本文選題：急性束縛應(yīng)激 + 慢性束縛應(yīng)激��； 參考：《華中科技大學(xué)》2006年博士論文

【摘要】： 第一部分急性和慢性束縛應(yīng)激對(duì)大鼠內(nèi)臟敏感性和神經(jīng)內(nèi)分泌的影響 目的：了解急性和慢性束縛應(yīng)激對(duì)大鼠內(nèi)臟敏感性以及神經(jīng)內(nèi)分泌反應(yīng)的影響特點(diǎn)和持續(xù)時(shí)間。方法：成年SD大鼠隨機(jī)分為對(duì)照組(沒有束縛應(yīng)激)，急性部分束縛應(yīng)激組(單次2h的束縛應(yīng)激)和慢性部分束縛應(yīng)激組(連續(xù)束縛應(yīng)激3天，每天2h)，，通過(guò)腹壁回撤反射(AWR)評(píng)分評(píng)估應(yīng)激前后大鼠內(nèi)臟對(duì)結(jié)直腸擴(kuò)張(CRD)的敏感性；并通過(guò)放免法檢測(cè)應(yīng)激前后不同時(shí)間點(diǎn)大鼠血漿促腎上腺皮質(zhì)激素(ACTH)以及皮質(zhì)酮(CORT)的水平。結(jié)果：(1)在擴(kuò)張壓力20，40mmHg時(shí)，急、慢性應(yīng)激組的AWR評(píng)分均顯著高于基礎(chǔ)水平(0d)(P＜0.05)，但在應(yīng)激后第7天(7d)AWR評(píng)分顯著下降(P＜0.05)。(2)急性應(yīng)激組血漿ACTH和CORT水平顯著高于正常對(duì)照組(P＜0.05)，但是應(yīng)激7日后，激素濃度下降至基礎(chǔ)水平。(3)慢性應(yīng)激組血漿ACTH和CORT水平在應(yīng)激組也顯著高于對(duì)照組，且激素水平在束縛7日后仍然維持在高水平(P＜0.05)。結(jié)論：急性和慢性束縛應(yīng)激都可以導(dǎo)致大鼠內(nèi)臟敏感性增高，但作用可能是短暫的。急性束縛應(yīng)激短暫顯著增加血漿ACTH和CORT的水平，而慢性束縛應(yīng)激可能長(zhǎng)期增加激素水平。神經(jīng)內(nèi)分泌激素的改變可以部分解釋束縛應(yīng)激導(dǎo)致內(nèi)臟高敏性的原因。 第二部分急性和慢性束縛應(yīng)激對(duì)外周5-羥色胺能系統(tǒng)的影響 目的：研究急性和慢性束縛應(yīng)激對(duì)大鼠結(jié)腸5-HT合成的限速酶、5-HT轉(zhuǎn)運(yùn)體、5-HT1A和5-HT4受體的影響。方法：成年SD大鼠隨機(jī)分為對(duì)照組(沒有接受部分束縛應(yīng)激(PRS))，急性部分束縛應(yīng)激組(A-PRS)，急性部分束縛應(yīng)激后恢復(fù)7日組(AR-PRS)，慢性部分束縛應(yīng)激組(C-PRS)，慢性束縛應(yīng)激后恢復(fù)7日組(CR-PRS)。采用RT-PCR半定量方法分別檢測(cè)以上各組大鼠近端以及遠(yuǎn)端結(jié)腸組織中的色氨酸羥化酶(TpH)，5-HT轉(zhuǎn)運(yùn)體(5-HTT)，5-HT1A以及5-HT4受體的mRNA表達(dá)。結(jié)果：(1)與對(duì)照組相比，C-PRS組以及CR-PRS組大鼠的TpH以及5-HTT mRNA顯著降低(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)急性束縛應(yīng)激組和急性束縛應(yīng)激后恢復(fù)7日組的TpH以及5-HTT的mRNA與正常組相比無(wú)顯著變化(P＞0.05)。(2)5-HT1A和5-HT4受體的mRNA在A-PRS和C-PRS組均顯著高于對(duì)照組(P＜0.05)。并且在恢復(fù)7日后，5-HT1A和5-HT4受體的mRNA仍維持在較高水平(P＜0.05)。結(jié)論：慢性束縛應(yīng)激抑制腸道5-HT合成、代謝的分子，而急性束縛應(yīng)激無(wú)顯著影響；急性和慢性束縛應(yīng)激均可以顯著增加腸道5-HT受體的表達(dá)。急、慢性束縛應(yīng)激對(duì)胃腸道功能失調(diào)的作用有不同機(jī)制。 第三部分替加色羅對(duì)急性束縛應(yīng)激導(dǎo)致內(nèi)臟敏感性和脊髓c-fos表達(dá)的影響 目的：研究替加色羅對(duì)急性束縛應(yīng)激導(dǎo)致的內(nèi)臟高敏感性和對(duì)脊髓相應(yīng)節(jié)段c-fos表達(dá)的影響。方法：本實(shí)驗(yàn)分為兩個(gè)部分。第一部分：雄性SD大鼠被隨機(jī)分為正常對(duì)照組(假性束縛應(yīng)激組)，束縛應(yīng)激后腹腔注射替加色羅0.3mg／kg、0.6mg／kg、1.2mg／kg組，或束縛應(yīng)激后腹腔注射溶酶0.3ml／kg、0.6ml／kg、1.2ml／kg對(duì)照組。通過(guò)觀察結(jié)直腸擴(kuò)張時(shí)(20，40，60，80mmHg)腹壁回撤反射評(píng)分，評(píng)估大鼠的內(nèi)臟敏感性。第二部分：雄性SD大鼠隨機(jī)分為束縛應(yīng)激組，束縛應(yīng)激后注射替加色羅1.2mg／kg組或溶媒1.2ml／kg組，以及正常對(duì)照組(假性束縛應(yīng)激組)，每組大鼠均接受重復(fù)的結(jié)直腸擴(kuò)張(60mmHg)2h。實(shí)驗(yàn)結(jié)束后取大鼠胸-腰段脊髓(T13，L1)和腰-骶段脊髓(L6，S1)，通過(guò)免疫組化方法檢測(cè)大鼠脊髓c-fos表達(dá)。結(jié)果：(1)各劑量溶酶對(duì)照應(yīng)激組的AWR評(píng)分在均顯著高于正常對(duì)照組(P＜0.05)；替加色羅應(yīng)激組AWR評(píng)分在各個(gè)劑量均顯著低于相應(yīng)劑量的溶酶應(yīng)激對(duì)照組，并且呈量-效依賴性改變(P＜0.05)。(2)與正常對(duì)照組相比，束縛應(yīng)激組或溶媒組可以顯著增加結(jié)直腸擴(kuò)張后胸腰段、以及腰骶段脊髓背角或側(cè)柱的c-fos表達(dá)(P＜0.05)，注射替加色羅后會(huì)顯著降低結(jié)直腸擴(kuò)張后胸腰段、以及腰骶段脊髓背角或側(cè)柱的c-fos表達(dá)(P＜0.05)。結(jié)論：替加色羅可以量-效依賴性降低急性應(yīng)激導(dǎo)致的內(nèi)臟感覺過(guò)敏；并且可以顯著降低應(yīng)激后結(jié)直腸擴(kuò)張導(dǎo)致的胸腰段、以及腰骶段脊髓內(nèi)臟輸入?yún)^(qū)域的c-fos表達(dá)。
[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.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類號(hào)】：R363

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