嗎啡依賴及戒斷大鼠中樞CCK-8基因表達的變化
發(fā)布時間：2019-05-07 04:33
【摘要】： 目的:藥物依賴(drug dependence)是一種慢性、復發(fā)性的腦疾病,可引起大腦發(fā)生病理性變化。在我國濫用的毒品主要是阿片類物質(zhì),阿片類物質(zhì)依賴包括身體依賴(表現(xiàn)為停藥后的戒斷癥狀)和精神依賴(表現(xiàn)為滿足感和重新用藥的渴求)。與阿片依賴及耐受相關(guān)的因素有內(nèi)源性阿片肽、阿片受體及其受體后信號轉(zhuǎn)導系統(tǒng)功能狀態(tài)的適應性改變等。膽囊收縮素(cholecystokinin,CCK)最早發(fā)現(xiàn)于腸道并作為胃腸激素調(diào)節(jié)膽囊收縮和胰腺的生長分泌,后來的研究發(fā)現(xiàn)CCK廣泛存在于中樞和周圍神經(jīng)系統(tǒng),作為神經(jīng)遞質(zhì)或調(diào)質(zhì)發(fā)揮重要作用。CCK-8是目前已知的作用最強的內(nèi)源性抗阿片肽,已有研究表明,CCK-8參與慢性嗎啡依賴及戒斷,但作用機制尚不清楚。本實驗通過觀察慢性嗎啡依賴及戒斷大鼠額葉皮質(zhì)、尾殼核、海馬CCK-8基因表達的變化,以探討CCK-8在嗎啡依賴及戒斷反應中可能的作用。 方法:健康雄性Wistar大鼠36只,體重180～200 g,隨機分組,每組12只,實驗分為:生理鹽水對照組(NS組)、嗎啡依賴組(MOR組)、納絡(luò)酮催促戒斷組(NAL組)。以劑量遞增法(10mg·kg-1, 20mg·kg-1, 30mg·kg-1,40mg·kg-1, 50mg·kg-1)連續(xù)5天皮下注射嗎啡建立嗎啡依賴模型,以納絡(luò)酮5mg·kg-1單次腹腔注射建立嗎啡依賴催促戒斷模型,觀察納絡(luò)酮催促戒斷癥狀并評價造模是否成功;納絡(luò)酮戒斷后1小時處死大鼠取額葉皮質(zhì)、尾殼核、海馬,每組取6只應用反轉(zhuǎn)錄聚合酶鏈反應(RT-PCR)檢測CCK-8 mRNA含量;每組另外6只應用免疫組化(IHC)檢測額葉皮質(zhì)、尾殼核、海馬CCK-8蛋白含量。用SPSS 11.5統(tǒng)計軟件對實驗數(shù)據(jù)進行分析,以P0.05為差異有顯著性。 結(jié)果: 1.嗎啡依賴動物模型的建立NS組和MOR組大鼠體重分別下降1.85±0.62g和2.63±0.8g,二者無顯著差異,而NAL組大鼠體重下降(13.67±2.5g)較MOR組明顯增加(P0.01);NS組和MOR組大鼠無一例發(fā)生跳躍反應,NAL組在注射納絡(luò)酮后的15min內(nèi)跳躍次數(shù)達到33.7±3.99次,與MOR組相比明顯增加(P0.01);注射納洛酮后,NAL組大鼠均出現(xiàn)濕狗樣震顫、不同程度的激惹、咬牙、流涎、腹瀉以及扭體,戒斷癥狀分值明顯高于NS和MOR組(P0.01),其中NS組和MOR組大鼠評分均為0分,而NAL組大鼠的評分達到32.7±3.62。表明成功建立了嗎啡身體依賴和納絡(luò)酮催促戒斷動物模型。 2.嗎啡依賴及戒斷大鼠額葉皮質(zhì)、尾殼核、海馬CCK-8 mRNA含量的變化 MOR組大鼠尾殼核CCK-8 mRNA含量(0.609±0.030)較NS組(0.458±0.062)顯著增高(P0.01),NAL組CCK-8 mRNA含量(0.437±0.059)較MOR組顯著降低(P0.01),NAL組CCK-8 mRNA含量與NS組之間無明顯差異(P0.05);額葉皮質(zhì)、海馬CCK-8 mRNA含量在各組之間無明顯差異(P0.05)。 3.嗎啡依賴及戒斷大鼠額葉皮質(zhì)、尾殼核、海馬CCK-8蛋白含量的變化 MOR組大鼠尾殼核CCK-8蛋白含量(1.300±0.067)較NS組(0.956±0.058)顯著增高(P0.01),NAL組CCK-8蛋白含量(0.956±0.084)較MOR組顯著降低(P0.01),NAL組CCK-8蛋白含量與NS組之間無明顯差異(P0.05);額葉皮質(zhì)、海馬CCK-8蛋白含量在各組之間無明顯差異(P0.05)。 結(jié)論: 1.慢性嗎啡依賴大鼠尾殼核CCK-8基因表達升高,納絡(luò)酮能夠翻轉(zhuǎn)此效應。 2.慢性嗎啡依賴及納絡(luò)酮催促戒斷大鼠,額葉皮質(zhì)、海馬CCK-8基因表達無變化。
[Abstract]:Objective: Drug dependence is a chronic and recurrent brain disease that can cause pathological changes in the brain. The drug abuse in our country is mainly opioids, and opioid dependence includes physical dependence (manifested as withdrawal symptoms after withdrawal) and mental dependence (manifested as a craving for satisfaction and remedication). The factors associated with opioid dependence and tolerance have endogenous opioid peptides, opioid receptors, and adaptive changes in the functional state of the signal transduction system after its receptor. Cholecystokinin (CCK) was first found in the intestinal tract and used as a gastrointestinal hormone to regulate the contraction of the gallbladder and the growth and secretion of the pancreas. CCK-8 is the most potent endogenous anti-opioid peptide currently known, and it has been shown that CCK-8 is involved in chronic morphine dependence and withdrawal, but the mechanism of action is not clear. The changes of the expression of CCK-8 gene in the frontal cortex, caudate nucleus and hippocampus of rats with chronic morphine dependence and withdrawal were observed to explore the possible role of CCK-8 in morphine dependence and withdrawal. Methods:36 healthy male Wistar rats were randomly divided into two groups: the normal saline control group (NS group), the morphine dependent group (MOR group) and the naloxone urging withdrawal group (NAL). The morphine dependence model was established by subcutaneous injection of morphine for 5 days in a dose-escalation method (10 mg 路 kg-1,20 mg 路 kg-1,30 mg 路 kg-1,40 mg 路 kg-1,50 mg 路 kg-1). The model was used to observe the effect of naloxone on the withdrawal symptoms and to evaluate the success of the model. One hour after the withdrawal of naloxone, the rats were sacrificed to take the cortex of the frontal lobe, the nucleus of the caudate shell and the hippocampus, and 6 of each group were used to detect the CCK-8 mRNA by reverse transcription polymerase chain reaction (RT-PCR). The content of the frontal cortex, the caudate nucleus and the CCK-8 protein in the frontal cortex, the caudate shell and the hippocampus were detected by immunohistochemistry (IHC) in each group. The data of the experimental data were analyzed by SPSS 11.5. The difference between the data and the experimental data was 0.05. The writing. Results:1. The weight of rats in NS group and MOR group decreased 1.85%, 0.62g and 2.63-0.8g, respectively. There was no significant difference between the two groups. In a case of a jump reaction, the number of hops in the NAL group was 33.7-3.99 times within 15 minutes after injection of naloxone, and compared with that of the MOR group (P0.01); after injection of naloxone, the rats of the nal group had a wet dog-like tremor, a different degree of agitation, a bite, a flow, The scores of salivation, diarrhea and torsion and withdrawal symptoms were significantly higher than that of NS and MOR (P0.01). The scores of rats in NS group and MOR group were 0, while the score of the rats in the NAL group was 3. 2.7 Figure 3.62. The successful establishment of morphine body-dependent and naloxone 2. Morphine-dependent and withdrawal rat frontal cortex, caudate nucleus, and hippocampus CC The content of CCK-8 mRNA in the rat tail shell of the modified MOR group (0.609-0.030) was significantly higher than that of the NS group (0.458-0.062) (P0.01), and the content of CCK-8 mRNA in the NAL group (0.437-0.059) was higher than that in the NS group (P 0.01). The levels of CCK-8 mRNA and CCK-8 mRNA in the frontal cortex and hippocampus in the frontal cortex and hippocampus were significantly lower than that in the NS group (P0.05). 3. There was no significant difference between morphine dependence and the frontal cortex of the withdrawal rats (P0.05). The content of CCK-8 (1.300-0.067) in the tail-shell nucleus and the content of the CCK-8 protein in the hippocampus was significantly higher than that in the NS group (0.956-0.058) (P0.01), and the content of the CCK-8 protein in the NAL group (0.956-0.067). There was no significant difference between the content of CCK-8 and the NS group (P <0.01), and there was no significant difference between the content of CCK-8 and the NS group (P0.05). white content There was no significant difference between the groups (P0.05). Conclusion:1. Chronic morphine dependence is large the expression of CCK-8 gene in the rat tail shell was increased, and the effect of naloxone could be reversed.2. Chronic morphine dependence and
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：D919
本文編號：2470776
[Abstract]:Objective: Drug dependence is a chronic and recurrent brain disease that can cause pathological changes in the brain. The drug abuse in our country is mainly opioids, and opioid dependence includes physical dependence (manifested as withdrawal symptoms after withdrawal) and mental dependence (manifested as a craving for satisfaction and remedication). The factors associated with opioid dependence and tolerance have endogenous opioid peptides, opioid receptors, and adaptive changes in the functional state of the signal transduction system after its receptor. Cholecystokinin (CCK) was first found in the intestinal tract and used as a gastrointestinal hormone to regulate the contraction of the gallbladder and the growth and secretion of the pancreas. CCK-8 is the most potent endogenous anti-opioid peptide currently known, and it has been shown that CCK-8 is involved in chronic morphine dependence and withdrawal, but the mechanism of action is not clear. The changes of the expression of CCK-8 gene in the frontal cortex, caudate nucleus and hippocampus of rats with chronic morphine dependence and withdrawal were observed to explore the possible role of CCK-8 in morphine dependence and withdrawal. Methods:36 healthy male Wistar rats were randomly divided into two groups: the normal saline control group (NS group), the morphine dependent group (MOR group) and the naloxone urging withdrawal group (NAL). The morphine dependence model was established by subcutaneous injection of morphine for 5 days in a dose-escalation method (10 mg 路 kg-1,20 mg 路 kg-1,30 mg 路 kg-1,40 mg 路 kg-1,50 mg 路 kg-1). The model was used to observe the effect of naloxone on the withdrawal symptoms and to evaluate the success of the model. One hour after the withdrawal of naloxone, the rats were sacrificed to take the cortex of the frontal lobe, the nucleus of the caudate shell and the hippocampus, and 6 of each group were used to detect the CCK-8 mRNA by reverse transcription polymerase chain reaction (RT-PCR). The content of the frontal cortex, the caudate nucleus and the CCK-8 protein in the frontal cortex, the caudate shell and the hippocampus were detected by immunohistochemistry (IHC) in each group. The data of the experimental data were analyzed by SPSS 11.5. The difference between the data and the experimental data was 0.05. The writing. Results:1. The weight of rats in NS group and MOR group decreased 1.85%, 0.62g and 2.63-0.8g, respectively. There was no significant difference between the two groups. In a case of a jump reaction, the number of hops in the NAL group was 33.7-3.99 times within 15 minutes after injection of naloxone, and compared with that of the MOR group (P0.01); after injection of naloxone, the rats of the nal group had a wet dog-like tremor, a different degree of agitation, a bite, a flow, The scores of salivation, diarrhea and torsion and withdrawal symptoms were significantly higher than that of NS and MOR (P0.01). The scores of rats in NS group and MOR group were 0, while the score of the rats in the NAL group was 3. 2.7 Figure 3.62. The successful establishment of morphine body-dependent and naloxone 2. Morphine-dependent and withdrawal rat frontal cortex, caudate nucleus, and hippocampus CC The content of CCK-8 mRNA in the rat tail shell of the modified MOR group (0.609-0.030) was significantly higher than that of the NS group (0.458-0.062) (P0.01), and the content of CCK-8 mRNA in the NAL group (0.437-0.059) was higher than that in the NS group (P 0.01). The levels of CCK-8 mRNA and CCK-8 mRNA in the frontal cortex and hippocampus in the frontal cortex and hippocampus were significantly lower than that in the NS group (P0.05). 3. There was no significant difference between morphine dependence and the frontal cortex of the withdrawal rats (P0.05). The content of CCK-8 (1.300-0.067) in the tail-shell nucleus and the content of the CCK-8 protein in the hippocampus was significantly higher than that in the NS group (0.956-0.058) (P0.01), and the content of the CCK-8 protein in the NAL group (0.956-0.067). There was no significant difference between the content of CCK-8 and the NS group (P <0.01), and there was no significant difference between the content of CCK-8 and the NS group (P0.05). white content There was no significant difference between the groups (P0.05). Conclusion:1. Chronic morphine dependence is large the expression of CCK-8 gene in the rat tail shell was increased, and the effect of naloxone could be reversed.2. Chronic morphine dependence and
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：D919
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