间歇缺氧大鼠模型的实验研究研究一：氧化应激、炎症反应的机制研究研究二：对甲状腺的相关影响

发布时间：2018-06-05 11:10

本文选题：阻塞性睡眠呼吸暂停低通气综合征 + 间歇缺氧　；参考：《兰州大学》2014年硕士论文

【摘要】：目的通过模拟阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者睡眠过程中反复发生的间歇低氧／再氧合过程,建立间歇缺氧模型,探讨间歇缺氧大鼠体内氧化应激、炎症反应的机制。方法将16只雄性SD大鼠进行称重后,随机分为两组,间歇缺氧(IH)组和对照(NC)组,每组8只,将其放入间歇缺氧模拟舱内,向舱内循环充入氮气、氧气和压缩空气,循环周期为120s,输入纯氮气30秒(模拟舱中氧浓度逐渐下降至6-7%),静息10秒(模拟舱中氧浓度维持在6-7%),输入纯氧气20秒(模拟舱中氧浓度逐渐上升至20-21%),输入压缩空气60秒(模拟舱中氧浓度波动在20.8±0.3%)。实验时间设计为每天8小时(9am-17pm),每周7天,持续4周,共28天。将对照组大鼠于每天相同时间置于相同规格的动物饲养舱内,并通入相同流量的空气。第29天处死大鼠后,用生化法测定大鼠血清MDA. SOD.GSH-PX水平,用放免法测定大鼠血清IL-6和TNF-a水平。结果1.血清MDA水平：正常对照组为5.09±0.51nmol/ml,IH组为5.74±0.38nmol/ml。与对照组比较,IH组大鼠血清MDA水平升高,且具有统计学差异(P=0.0110.05)。2.血清SOD水平：正常对照组为67.21±6.68U/ml,IH组56.70±6.34U/ml。与对照组相比,IH组大鼠血请SOD水平降低,且具有统计学差异(P=0.006＜0.05)。3.血清GSH-PX水平：正常对照组为811.50±14.90U/ml,IH组779.16±30.27U/ml。与对照组相比,IH组大鼠血清GSH-PX水平降低,且具有统计学差异(P=0.017＜0.05)。4.血清IL-6水平：正常对照组为166.04±30.92pg/ml,IH组为211.01±33.24pg/ml.与对照组比较,IH组大鼠血清IL-6水平升高,且具有统计学差异(P=0.014＜0.05)。4.血清TNF-a水平：正常对照组为59.11±10.07pg/ml,IH组74.22±7.68pg/ml.与对照组相比,IH组大鼠血请TNF-a水平升高,且具有统计学差异(P=0.005＜0.05)。结论1.间歇缺氧条件下能导致大鼠血清MDA水平升高,而血清SOD和GSH水平降低,提示间歇缺氧大鼠体内存在氧化应激状态,间歇缺氧可以导致大鼠体内抗氧化能力的下降；2.间歇缺氧条件下能导致大鼠血清TNF-a和IL-6水平升高,提示间歇缺氧大鼠体内存在炎症反应。目的通过模拟阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者睡眠过程中反复发生的间歇低氧／再氧合过程,建立间歇缺氧模型,探讨间歇缺氧对大鼠甲状腺激素、甲状腺超微结构及对大鼠甲状腺Peroxiredoxin5表达的影响。方法将16只雄性SD大鼠进行称重后,随机分为两组,间歇缺氧(IH)组和对照(NC)组,每组8只,将其放入间歇缺氧模拟舱内,向舱内循环充入氮气、氧气和压缩空气,循环周期为120s,输入纯氮气30秒(模拟舱中氧浓度逐渐下降至6-7%),静息10秒(模拟舱中氧浓度维持在6-7%),输入纯氧气20秒(模拟舱中氧浓度逐渐上升至20-21%),输入压缩空气60秒(模拟舱中氧浓度波动在20.8±0.3%)。实验时间设计为每天8小时(9am-17pm),每周7天,持续4周,共28天。将对照组大鼠于每天相同时间置于相同规格的动物饲养舱内,并通入相同流量的空气。第29天处死大鼠后,用放免法测定大鼠血清FT3、FT4、TSH水平,于电镜下观察大鼠甲状腺超微结构的改变,用免疫组化方法测定大鼠甲状腺中Peroxiredoxin5表达。结果1.血清FT3水平：正常对照组为0.40±0.17ng/dl,IH组为0.46±0.19ng/dl。与对照组比较,IH组大鼠血清FT3水平升高,但不具有统计学差异(P=0.5210.05)。2.血清FT4水平：正常对照组为4.20±0.92ng/dl,IH组2.98±0.64ng/dl。与对照组相比,IH组大鼠血清FT4水平降低,且具有统计学差异(P=0.0090.05)。3.血清TSH水平：正常对照组为12.34±1.89uIU/ml,IH组9.36±1.17uIU/ml。与对照组相比,IH组大鼠血清TSH水平降低,且具有统计学差异(P=0.0030.05)。4.IH组大鼠甲状腺超微结构：整体表现为缺氧改变。5.甲状腺PRDX5的阳性染色主要位于甲状腺细胞的胞浆中,大鼠甲状腺Peroxiredoxin5表达：正常对照组的平均光密度为0.22±0.01,CIH组的平均光密度为0.30±0.01。与对照组比较,IH组大鼠甲状腺Peroxiredoxin5表达升高且差异具有统计学意义(P=0.0020.05)。结论1.间歇缺氧条件下能导致大鼠血清FT4、TSH降低,提示了间歇缺氧可以引起大鼠下丘脑-垂体-甲状腺轴及甲状腺本身功能的下降或异常,从而导致甲状腺激素水平发生改变；2.间歇缺氧条件下能导致大鼠甲状腺超微结构发生缺氧改变；3.间歇缺氧条件下能导致大鼠甲状腺中PRDX5阳性表达升高,提示了间歇缺氧可使大鼠甲状腺抗氧化能力增加,间接提示了间歇缺氧大鼠甲状腺存在氧化应激状态。
[Abstract]:Objective to simulate intermittent hypoxia / reoxygenation during the sleep process of patients with obstructive sleep apnea hypopnea syndrome (OSAHS), to establish an intermittent hypoxia model, and to explore the mechanism of oxidative stress and inflammation in intermittent hypoxia rats.
Methods 16 male SD rats were weighed. They were randomly divided into two groups, intermittent hypoxia (IH) group and control (NC) group, 8 in each group. They were put into the intermittent hypoxia simulated cabin and filled the cabin with nitrogen, oxygen and compressed air, the cycle period was 120s, and the pure nitrogen gas was entered for 30 seconds (the oxygen concentration in the simulated cabin gradually dropped to 6-7%) and resting for 10 seconds (simulation). The oxygen concentration in the cabin was maintained at 6-7%), the pure oxygen was input for 20 seconds (the oxygen concentration in the simulated cabin was gradually increased to 20-21%), and the compressed air was input for 60 seconds (the oxygen concentration in the simulated cabin was 20.8 + 0.3%). The experimental time was designed to be 8 hours a day (9am-17pm), 7 days a week, and 4 weeks for 28 days. The control group was placed in the same specification at the same time of the day. The animals were fed with the same flow of air. After twenty-ninth days of death, the rat serum MDA. SOD.GSH-PX level was measured by biochemical method, and the serum levels of IL-6 and TNF-a were measured by radioimmunoassay.
Results 1. serum MDA level: the normal control group was 5.09 + 0.51nmol/ml, and the IH group was 5.74 + 0.38nmol/ml. compared with the control group. The serum MDA level of the IH group was higher, and the serum SOD level of.2. was statistically different (P=0.0110.05).2. serum SOD level: the normal control group was 67.21 + 6.68U/ml, and the IH group 56.70 + 6.34U/ml. was compared with the control group. Decrease, and have statistical difference (P=0.006 < 0.05).3. serum GSH-PX level: normal control group was 811.50 + 14.90U/ml, IH group 779.16 + 30.27U/ml. compared with control group, IH group rats serum GSH-PX level decreased, and there was statistical difference (P=0.017 < 0.05).4. serum IL-6 level: normal control group was 166.04 + 30.92pg/ml, 211.01 group was 211.01 Compared with the control group, the serum IL-6 level of the rats in the IH group was increased, and the serum TNF-a level of.4. was statistically different (P=0.014 < 0.05).4. serum TNF-a level: the normal control group was 59.11 + 10.07pg/ml, and the IH group was 74.22 + 7.68pg/ml. compared with the control group, the TNF-a level of the IH group was higher, and there was a statistical difference (P=0.005 < 0.05).
Conclusion 1. intermittent hypoxia can lead to the increase of serum MDA level in rats, and the decrease of serum SOD and GSH levels, suggesting that there is oxidative stress in rats with intermittent hypoxia. Intermittent hypoxia can lead to the decrease of antioxidant capacity in rats. 2. intermittent hypoxia can lead to the increase of serum level of TNF-a and IL-6 in rats, suggesting intermittent deficiency. There are inflammatory reactions in the rats.
Objective to simulate intermittent hypoxia / reoxygenation during the sleep process of obstructive sleep apnea hypopnea syndrome (OSAHS), and to establish an intermittent hypoxia model to explore the effect of intermittent hypoxia on thyroid hormone, thyroid ultrastructure and the expression of thyroid gland Peroxiredoxin5 in rats.
Methods 16 male SD rats were weighed. They were randomly divided into two groups, intermittent hypoxia (IH) group and control (NC) group, 8 in each group. They were put into the intermittent hypoxia simulated cabin and filled the cabin with nitrogen, oxygen and compressed air, the cycle period was 120s, and the pure nitrogen gas was entered for 30 seconds (the oxygen concentration in the simulated cabin gradually dropped to 6-7%) and resting for 10 seconds (simulation). The oxygen concentration in the cabin was maintained at 6-7%), the pure oxygen was input for 20 seconds (the oxygen concentration in the simulated cabin was gradually increased to 20-21%), and the compressed air was input for 60 seconds (the oxygen concentration in the simulated cabin was 20.8 + 0.3%). The experimental time was designed to be 8 hours a day (9am-17pm), 7 days a week, and 4 weeks for 28 days. The control group was placed in the same specification at the same time of the day. The animals were fed with the same flow of air. After twenty-ninth days of death, the rat serum FT3, FT4, TSH levels were measured by radioimmunoassay. The ultrastructural changes of the thyroid gland were observed under the electron microscope, and the expression of Peroxiredoxin5 in the thyroid gland was measured by immunohistochemistry.
Results 1. serum FT3 level: the normal control group was 0.40 + 0.17ng/dl, and the IH group was 0.46 + 0.19ng/dl. compared with the control group. The serum FT3 level of the IH group was higher, but there was no statistical difference (P=0.5210.05).2. serum FT4 level: the normal control group was 4.20 + 0.92ng/dl, and the IH group 2.98 + 0.64ng/dl. was compared with the control group. Decrease, and have statistical difference (P=0.0090.05).3. serum TSH level: the normal control group was 12.34 + 1.89uIU/ml, the IH group 9.36 + 1.17uIU/ml. and the control group compared with the control group, the serum TSH level of the IH group decreased, and there was statistical difference (P=0.0030.05).4.IH group rat thyroid ultrastructure: the overall performance was the hypoxia to change.5. thyroid PRDX5. The positive staining was mainly located in the cytoplasm of thyroid cells. The expression of Peroxiredoxin5 in the thyroid gland was 0.22 + 0.01 in the normal control group, and the average light density in the CIH group was 0.30 + 0.01. compared with the control group. The thyroid Peroxiredoxin5 expression in the IH group was higher and the difference was statistically significant (P=0.0020.05).
Conclusion 1. intermittent hypoxia can lead to the decrease of FT4 and TSH in rat serum, suggesting that intermittent hypoxia can cause the decrease or abnormal function of hypothalamus pituitary thyroid axis and thyroid itself, which leads to the change of thyroid hormone level, and 2. intermittent hypoxia can lead to hypoxic ultrastructural anoxia in rats. 3. intermittent hypoxia can lead to the increase of PRDX5 positive expression in the thyroid gland of rats, suggesting that intermittent hypoxia can increase the antioxidant capacity of thyroid in rats and indirectly suggest that there is oxidative stress in the thyroid gland of rats with intermittent hypoxia.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R766

【参考文献】

间歇缺氧大鼠模型的实验研究 研究一：氧化应激、炎症反应的机制研究 研究二：对甲状腺的相关影响

间歇缺氧大鼠模型的实验研究研究一：氧化应激、炎症反应的机制研究研究二：对甲状腺的相关影响