色氨酸对应激小鼠抗氧化能力和行为的影响
发布时间：2018-10-14 09:12
【摘要】：本文研究了色氨酸对应激小鼠氧化还原状态、免疫功能和行为的影响，通过灌胃不同剂量色氨酸(Trp)，观察其对小鼠下丘脑单胺类神经递质、氧化还原状态、免疫功能及行为和相关基因等调节作用，探讨其作用机制。 选用无特定病原体(SPF)健康昆明种雄性小鼠，体重为25-30g。环磷酰胺应激模型：实验期24天，采用笼养，12h光照，自由饮水与采食，操作符合相关动物实验法规。80只小鼠预饲一周后按照体重随机分为4组，每组20只：空白组(NC组)，应激模型组(CTX组)，应激模型-低剂量色氨酸组(CTX-L组)，应激模型-高剂量色氨酸组(CTX-H组)。预饲一周后，NC组1-3日每天腹腔注射生理盐水10mg/kg·bw，4-17日每天灌胃生理盐水10mg/kg·bw；CTX组1-3日每天腹腔注射浓度100mg/kg·bw的环磷酰胺，4-17日每天灌胃生理盐水10mg/kg·bw；CTX-L组1-3日每天腹腔注射浓度100mg/kg·bw的环磷酰胺，4-17日每天灌胃浓度为100mg/kg·bw的Trp；CTX-H组1-3日每天腹腔注射浓度100mg/kg·bw的环磷酰胺，4-17日每天灌胃浓度为200mg/kg·bw的Trp。可的松应激模型：60只SPF级雄性昆明种小鼠预饲一周后按照体重随机分为4组，每组15只：空白对照组(C组)，应激模型组(M组)，应激模型-低剂量色氨酸组(M-L组)，应激模型-高剂量色氨酸组(M-H组)。预饲一周后，C组及M组小鼠灌胃生理盐水10mg/kg·bw，M-L组灌胃100mg/kg·bw的色氨酸，M-H组灌胃200mg/kg·bw的色氨酸，，每日一次，连续14日。第8日给药后30min按10mg/kg·bw给予空白对照组小鼠腹腔注射生理盐水，其余各组均注射100mg/kg·bw氢化可的松琥珀酸钠。 实验结果：(1)环磷酰胺应激小鼠体质量增加速度、胸腺指数较对照组小鼠，显著下降(P0.05)，色氨酸有助于其体重和胸腺指数恢复，其中低剂量色氨酸效果更显著(P0.05)；(2)应激可导致小鼠血浆和组织氧化还原状态失衡，抗氧化能力下降，灌胃色氨酸有助于提高其抗氧化能力，根据不同组织，效果有所不同；高剂量色氨酸对于提高免疫能力效果更显著；(3)应激小鼠脑内5-羟色胺(Trp)含量下降，去甲肾上腺素(NE)水平上升，学习记忆能力和空间搜索能力显著下降(P0.05)；灌胃色氨酸使5-羟色胺和去甲肾上腺素水平得到改善，学习记忆能力和空间搜索能力得以恢复，低剂量色氨酸效果更好；(4)Real-time PCR结果显示，环磷酰胺应激小鼠下丘脑吲哚胺2,3-双加氧酶(IDO)表达上调，而肠道IDO表达量无显著性差异(P0.05)。可的松应激小鼠下丘脑内色氨酸羟化酶-2(TPH-2)表达显著上调(P0.05)，5-HT_(2A)、5-HT_7表达显著下调(P0.05)，从而导致小鼠认知能力下降。通过灌胃色氨酸可改善上述基因的表达异常。 结论：色氨酸有助于提高小鼠血浆和组织的抗氧化水平，且在一定程度上增强了机体免疫功能。适量色氨酸可以有效改善应激小鼠相关行为学评分，其机制可能与下丘脑5-HT和NE的含量有关，并通过TPH-2、5-HT_(2A)、5-HT_7基因的表达量进行调控。
[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.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R151.2
本文编号：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.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R151.2
【参考文献】
相关期刊论文 前10条
1 张建东,侯一平,宋焱峰,张朗,王德贵;大豆磷脂酰胆碱提高幼年大鼠学习记忆能力及海马突触素的表达[J];第四军医大学学报;2005年09期
2 王大慧,韦萍,欧阳平凯;D-色氨酸研究进展[J];化工进展;2002年02期
3 畅洪f;梁吉春;石任兵;程可建;孙文燕;;枳术宽中胶囊对嗅球损毁大鼠抑郁症模型的抗抑郁作用研究[J];北京中医药大学学报;2009年08期
4 耿义群,苏敏,徐小虎;硒-谷胱甘肽过氧化物酶在脑抗氧化损伤中的研究进展[J];脑与神经疾病杂志;2004年03期
5 赵弋清,罗霞,陈东辉,余梦瑶,杨志荣;不同剂量环磷酰胺诱导正常小鼠免疫抑制的对比研究[J];免疫学杂志;2005年S1期
6 万福生，南国华;高苯丙氨酸血症大鼠脑内单胺类递质的研究[J];生物化学与生物物理进展;1995年04期
7 杨秀丽;张文高;郑广娟;;幼龄ApoE基因敲除小鼠认知功能障碍的行为学观察[J];世界中西医结合杂志;2008年01期
8 崔芹 ,崔山;色氨酸营养研究进展[J];中国饲料;2003年15期
9 刘能保,李辉,刘向前,孙臣友,陈少容,张敏海,刘少纯,汪薇曦;慢性复合应激增强大鼠空间学习和记忆能力[J];生理学报;2004年05期
10 代国杰;朱建津;高琳琳;单立娟;;褪黑素对小鼠抗氧化作用的影响[J];食品工业科技;2010年03期
本文编号：2270023
本文链接：https://www.wllwen.com/yixuelunwen/yufangyixuelunwen/2270023.html
