色氨酸對(duì)應(yīng)激小鼠抗氧化能力和行為的影響
發(fā)布時(shí)間：2018-10-14 09:12
【摘要】：本文研究了色氨酸對(duì)應(yīng)激小鼠氧化還原狀態(tài)、免疫功能和行為的影響，通過灌胃不同劑量色氨酸(Trp)，觀察其對(duì)小鼠下丘腦單胺類神經(jīng)遞質(zhì)、氧化還原狀態(tài)、免疫功能及行為和相關(guān)基因等調(diào)節(jié)作用，探討其作用機(jī)制。 選用無特定病原體(SPF)健康昆明種雄性小鼠，體重為25-30g。環(huán)磷酰胺應(yīng)激模型：實(shí)驗(yàn)期24天，采用籠養(yǎng)，12h光照，自由飲水與采食，操作符合相關(guān)動(dòng)物實(shí)驗(yàn)法規(guī)。80只小鼠預(yù)飼一周后按照體重隨機(jī)分為4組，每組20只：空白組(NC組)，應(yīng)激模型組(CTX組)，應(yīng)激模型-低劑量色氨酸組(CTX-L組)，應(yīng)激模型-高劑量色氨酸組(CTX-H組)。預(yù)飼一周后，NC組1-3日每天腹腔注射生理鹽水10mg/kg·bw，4-17日每天灌胃生理鹽水10mg/kg·bw；CTX組1-3日每天腹腔注射濃度100mg/kg·bw的環(huán)磷酰胺，4-17日每天灌胃生理鹽水10mg/kg·bw；CTX-L組1-3日每天腹腔注射濃度100mg/kg·bw的環(huán)磷酰胺，4-17日每天灌胃濃度為100mg/kg·bw的Trp；CTX-H組1-3日每天腹腔注射濃度100mg/kg·bw的環(huán)磷酰胺，4-17日每天灌胃濃度為200mg/kg·bw的Trp�？傻乃蓱�(yīng)激模型：60只SPF級(jí)雄性昆明種小鼠預(yù)飼一周后按照體重隨機(jī)分為4組，每組15只：空白對(duì)照組(C組)，應(yīng)激模型組(M組)，應(yīng)激模型-低劑量色氨酸組(M-L組)，應(yīng)激模型-高劑量色氨酸組(M-H組)。預(yù)飼一周后，C組及M組小鼠灌胃生理鹽水10mg/kg·bw，M-L組灌胃100mg/kg·bw的色氨酸，M-H組灌胃200mg/kg·bw的色氨酸，，每日一次，連續(xù)14日。第8日給藥后30min按10mg/kg·bw給予空白對(duì)照組小鼠腹腔注射生理鹽水，其余各組均注射100mg/kg·bw氫化可的松琥珀酸鈉。 實(shí)驗(yàn)結(jié)果：(1)環(huán)磷酰胺應(yīng)激小鼠體質(zhì)量增加速度、胸腺指數(shù)較對(duì)照組小鼠，顯著下降(P0.05)，色氨酸有助于其體重和胸腺指數(shù)恢復(fù)，其中低劑量色氨酸效果更顯著(P0.05)；(2)應(yīng)激可導(dǎo)致小鼠血漿和組織氧化還原狀態(tài)失衡，抗氧化能力下降，灌胃色氨酸有助于提高其抗氧化能力，根據(jù)不同組織，效果有所不同；高劑量色氨酸對(duì)于提高免疫能力效果更顯著；(3)應(yīng)激小鼠腦內(nèi)5-羥色胺(Trp)含量下降，去甲腎上腺素(NE)水平上升，學(xué)習(xí)記憶能力和空間搜索能力顯著下降(P0.05)；灌胃色氨酸使5-羥色胺和去甲腎上腺素水平得到改善，學(xué)習(xí)記憶能力和空間搜索能力得以恢復(fù)，低劑量色氨酸效果更好；(4)Real-time PCR結(jié)果顯示，環(huán)磷酰胺應(yīng)激小鼠下丘腦吲哚胺2,3-雙加氧酶(IDO)表達(dá)上調(diào)，而腸道IDO表達(dá)量無顯著性差異(P0.05)�？傻乃蓱�(yīng)激小鼠下丘腦內(nèi)色氨酸羥化酶-2(TPH-2)表達(dá)顯著上調(diào)(P0.05)，5-HT_(2A)、5-HT_7表達(dá)顯著下調(diào)(P0.05)，從而導(dǎo)致小鼠認(rèn)知能力下降。通過灌胃色氨酸可改善上述基因的表達(dá)異常。 結(jié)論：色氨酸有助于提高小鼠血漿和組織的抗氧化水平，且在一定程度上增強(qiáng)了機(jī)體免疫功能。適量色氨酸可以有效改善應(yīng)激小鼠相關(guān)行為學(xué)評(píng)分，其機(jī)制可能與下丘腦5-HT和NE的含量有關(guān)，并通過TPH-2、5-HT_(2A)、5-HT_7基因的表達(dá)量進(jìn)行調(diào)控。
[Abstract]:The effects of tryptophan on redox state, immune function and behavior in stressed mice were studied. The effects of tryptophan (Trp), on monoamine neurotransmitters and redox state in hypothalamus of mice were observed by intragastric administration of tryptophan (Trp),. Immune function, behavior and related genes were regulated and its mechanism was discussed. Healthy Kunming male mice without specific pathogen (SPF) were selected and their body weight was 25-30 g. Cyclophosphamide stress model: the experimental period was 24 days, the mice were treated with cage, 12h light, drinking water and feeding freely, and the operation was in accordance with the relevant animal experimental regulations. 80 mice were randomly divided into 4 groups according to their body weight after one week of prefeeding. There were 20 rats in each group: blank group (NC group), stress model group (CTX group), stress-low dose tryptophan group (CTX-L group), stress-high dose tryptophan group (CTX-H group). After one week of prefeeding, NC group received intraperitoneal injection of normal saline 10mg/kg bw,4-17 daily on 1-3 days, 10mg/kg bw;CTX group 1 to 3 days after intraperitoneal injection of cyclophosphamide with 100mg/kg bw concentration, and normal saline 10mg/kg bw; daily on 4-17 days. CTX-L group was intraperitoneally injected with cyclophosphamide of 100mg/kg bw concentration on 1-3 days, Trp;CTX-H group with 100mg/kg bw concentration on 4-17 days, Trp;CTX-H group with daily 100mg/kg bw concentration of cyclophosphamide on 1-3 days and Trp. with 200mg/kg bw concentration daily on 4-17 days. Cortisone stress model: 60 male Kunming mice of SPF grade were randomly divided into 4 groups according to their body weight after one week of prefeeding. There were 15 rats in each group: blank control group (C group), stress model group (M group), stress model-low dose tryptophan group (M-L group), stress model-high dose tryptophan group (M-H group). After one week of prefeeding, mice in group C and group M were fed with tryptophan of 100mg/kg bw in 10mg/kg bw,M-L group and 200mg/kg bw in M-H group, once a day for 14 days. After the 8th day of administration, 30min was injected intraperitoneally with normal saline according to 10mg/kg bw in blank control group, and other groups were injected with 100mg/kg bw hydrocortisone sodium succinate. The results showed that: (1) the weight of mice under cyclophosphamide stress increased and the thymus index decreased significantly compared with the control group (P0.05). Tryptophan contributed to the recovery of body weight and thymus index, and the effect of low dose tryptophan was more significant (P0.05). (2) stress could lead to the imbalance of redox state in plasma and tissue, the decrease of antioxidant capacity, and the increase of antioxidative ability by oral administration of tryptophan, which had different effects according to different tissues. The effect of high dose tryptophan on improving immune ability was more significant. (3) the content of 5-hydroxytryptamine (Trp) decreased, the level of norepinephrine (NE) increased, the ability of learning and memory and the ability of spatial search decreased significantly in stress mice (P0.05). Oral administration of tryptophan improved serotonin and norepinephrine levels, improved learning and memory ability and spatial search ability, and improved the effect of low dose tryptophan. (4) Real-time PCR results showed that, The expression of (IDO) in hypothalamus was up-regulated in cyclophosphamide stress mice, but there was no significant difference in the expression of IDO in intestinal tract (P0.05). The expression of tryptophan hydroxylase 2 (TPH-2) in hypothalamus of cortisone stressed mice was significantly up-regulated (P0.05), the expression of 5-HT2A and 5-HT_7 were significantly down-regulated (P0.05), which resulted in the decrease of cognitive ability of mice. The abnormal expression of these genes could be improved by oral administration of tryptophan. Conclusion: tryptophan can improve the level of antioxidation in plasma and tissue of mice, and enhance the immune function to some extent. Proper amount of tryptophan can effectively improve the related behavioral score of stress mice, and its mechanism may be related to the content of 5-HT and NE in hypothalamus and regulated by the expression of TPH-2,5-HT_ (2A) and 5-HT_7 genes.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R151.2
本文編號(hào)：2270023
[Abstract]:The effects of tryptophan on redox state, immune function and behavior in stressed mice were studied. The effects of tryptophan (Trp), on monoamine neurotransmitters and redox state in hypothalamus of mice were observed by intragastric administration of tryptophan (Trp),. Immune function, behavior and related genes were regulated and its mechanism was discussed. Healthy Kunming male mice without specific pathogen (SPF) were selected and their body weight was 25-30 g. Cyclophosphamide stress model: the experimental period was 24 days, the mice were treated with cage, 12h light, drinking water and feeding freely, and the operation was in accordance with the relevant animal experimental regulations. 80 mice were randomly divided into 4 groups according to their body weight after one week of prefeeding. There were 20 rats in each group: blank group (NC group), stress model group (CTX group), stress-low dose tryptophan group (CTX-L group), stress-high dose tryptophan group (CTX-H group). After one week of prefeeding, NC group received intraperitoneal injection of normal saline 10mg/kg bw,4-17 daily on 1-3 days, 10mg/kg bw;CTX group 1 to 3 days after intraperitoneal injection of cyclophosphamide with 100mg/kg bw concentration, and normal saline 10mg/kg bw; daily on 4-17 days. CTX-L group was intraperitoneally injected with cyclophosphamide of 100mg/kg bw concentration on 1-3 days, Trp;CTX-H group with 100mg/kg bw concentration on 4-17 days, Trp;CTX-H group with daily 100mg/kg bw concentration of cyclophosphamide on 1-3 days and Trp. with 200mg/kg bw concentration daily on 4-17 days. Cortisone stress model: 60 male Kunming mice of SPF grade were randomly divided into 4 groups according to their body weight after one week of prefeeding. There were 15 rats in each group: blank control group (C group), stress model group (M group), stress model-low dose tryptophan group (M-L group), stress model-high dose tryptophan group (M-H group). After one week of prefeeding, mice in group C and group M were fed with tryptophan of 100mg/kg bw in 10mg/kg bw,M-L group and 200mg/kg bw in M-H group, once a day for 14 days. After the 8th day of administration, 30min was injected intraperitoneally with normal saline according to 10mg/kg bw in blank control group, and other groups were injected with 100mg/kg bw hydrocortisone sodium succinate. The results showed that: (1) the weight of mice under cyclophosphamide stress increased and the thymus index decreased significantly compared with the control group (P0.05). Tryptophan contributed to the recovery of body weight and thymus index, and the effect of low dose tryptophan was more significant (P0.05). (2) stress could lead to the imbalance of redox state in plasma and tissue, the decrease of antioxidant capacity, and the increase of antioxidative ability by oral administration of tryptophan, which had different effects according to different tissues. The effect of high dose tryptophan on improving immune ability was more significant. (3) the content of 5-hydroxytryptamine (Trp) decreased, the level of norepinephrine (NE) increased, the ability of learning and memory and the ability of spatial search decreased significantly in stress mice (P0.05). Oral administration of tryptophan improved serotonin and norepinephrine levels, improved learning and memory ability and spatial search ability, and improved the effect of low dose tryptophan. (4) Real-time PCR results showed that, The expression of (IDO) in hypothalamus was up-regulated in cyclophosphamide stress mice, but there was no significant difference in the expression of IDO in intestinal tract (P0.05). The expression of tryptophan hydroxylase 2 (TPH-2) in hypothalamus of cortisone stressed mice was significantly up-regulated (P0.05), the expression of 5-HT2A and 5-HT_7 were significantly down-regulated (P0.05), which resulted in the decrease of cognitive ability of mice. The abnormal expression of these genes could be improved by oral administration of tryptophan. Conclusion: tryptophan can improve the level of antioxidation in plasma and tissue of mice, and enhance the immune function to some extent. Proper amount of tryptophan can effectively improve the related behavioral score of stress mice, and its mechanism may be related to the content of 5-HT and NE in hypothalamus and regulated by the expression of TPH-2,5-HT_ (2A) and 5-HT_7 genes.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R151.2
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