褪黑素对慢性间歇缺氧大鼠海马神经元凋亡相关基因bcl-2、bax表达及氧化应激的影响
发布时间：2018-11-09 08:48
【摘要】：研究目的 OSAHS是以反复发生上气道完全或部分阻塞导致呼吸暂停或低通气,其突出的病理生理过程是夜间反复发作性低氧/复氧。反复发作性低氧/复氧-即慢性间歇缺氧可以导致多器官损害,神经系统损害尤其常见,例如记忆力下降,认知功能受损等。大量海马组织神经元凋亡是OSAHS患者学习和认知功能损害的关键,活性氧在慢性间歇缺氧对海马神经元长期损害中起关键作用。 持续气道正压通气(CPAP)是目前治疗OSAHS中疗效最确切的一种方法,但仍有部分患者白天嗜睡改善不明显,有部分患者不能耐受呼吸机而放弃治疗,尤其是轻中度睡眠呼吸暂停患者顺从性差。 慢性间歇缺氧是OSAHS患者最重要的病理生理特征,因此用OSAHS动物模型探讨慢性间歇缺氧对靶器官损害的机制及相应的干预方法目前已成为临床研究的热点。为此,我们选用慢性间歇缺氧大鼠模型研究OSAHS患者存在的缺氧/再复氧病理生理过程,同时应用抗氧化剂褪黑素进行治疗,观察慢性间歇缺氧海马组织中氧化应激指标MDA、SOD及bcl-2,bax蛋白的表达及海马神经元凋亡情况。探讨抗氧化剂MT在CIH大鼠模型中的治疗作用及其相关作用机制。 方法 将30只2月龄SD健康雄性大鼠随机分为常氧+2%的乙生理盐水腹腔注射对照组(A, n=10);慢性间歇缺氧+2%的醇理盐水腹腔注射组(B, n=10);慢性间歇缺氧+2%的乙醇黑素腹腔注射组(C, n=10)。采用化学比色法检测海马组织化应激指标超:超氧化物歧化酶(SOD)、丙二醛(MDA)水平;用免疫组化学SP法检测海马组织凋亡指标:bax,bcl-2蛋白的表达水平;采用TUNEL法检测海马组织神经元凋亡率。 结果 MT治疗组海马组织中MDA水平[(0.6760±0.0687)nmol/mg蛋白]显著低于正常对照组[(1.1±0.16)nmol/mg蛋白]和CIH组[ (1.7±0.17)nmol/mg蛋白)],各组大鼠海马组织中MDA水平,组间比较差异有统计学意(F=12.876,P0.01)。MT治疗组大鼠海马组织中SOD活性[(46.92±5.66)U/mg蛋白明显高于CIH组([32.288±2.984)U/mg蛋白],但低于正常对照组[(64.28±5.29)U/mg蛋白],各组间比较均有统计学差异(F=11.2525,P0.01)。MT治疗组的bcl-2蛋白的阳性表达细胞灰度值(0.220±0.029)较CIH组(0.0968±0.0173)及正常组(0.142±0.0178)明显增高F=64.138,P0.0差异有统计学意义。MT治疗组组的bax蛋白的阳性表达灰度值(0.135±0.00688)较CIH组(0.222±0.0256)及正常对照组降低(0.176±0.012),F=34.759,P0.05),差异有统计学意义。MT治疗组的凋亡细胞阳性表达细胞灰度值(0.1930±0.0202)较CIH组(0.335±0.0532)明显降低,但较正常对照(0.0675±0.0125)高,(F=64.138 P0.05)差异有统计学意义 结论 褪黑素对慢性间歇缺氧大鼠海马神经元凋亡有明显的抑制及抗氧化应激作用,bcl-2表达降低和bax表达增加可能是其凋亡机制之一
[Abstract]:Objective OSAHS is caused by apnea or hypopnea caused by repeated complete or partial obstruction of upper airway. The prominent pathophysiological process of OSAHS is repeated paroxysmal hypoxia / reoxygenation at night. Recurrent hypoxic / reoxygenation-chronic intermittent hypoxia can lead to multiple organ damage, especially in the nervous system, such as memory loss, cognitive impairment and so on. A large number of hippocampal neurons apoptosis is the key to learning and cognitive impairment in patients with OSAHS. Reactive oxygen species (Ros) play a key role in the long-term damage of hippocampal neurons caused by chronic intermittent hypoxia. Continuous positive airway pressure ventilation (CPAP) is the most effective method in the treatment of OSAHS, but there are still some patients who are unable to tolerate ventilator and give up treatment. Especially mild to moderate sleep apnea patients with poor compliance. Chronic intermittent hypoxia is the most important pathophysiological feature in patients with OSAHS. Therefore, the mechanism of chronic intermittent hypoxia on target organ damage and the corresponding intervention methods have become the focus of clinical research. Therefore, we selected the chronic intermittent hypoxia rat model to study the pathophysiological process of hypoxia / reoxygenation in patients with OSAHS, and treated with antioxidant melatonin, and observed the oxidative stress index MDA, in the hippocampus of chronic intermittent hypoxia. Expression of SOD and bcl-2,bax protein and apoptosis of hippocampal neurons. To investigate the therapeutic effect of antioxidant MT in CIH rat model and its related mechanism. Methods Thirty 2-month-old SD healthy male rats were randomly divided into 2% normoxic saline group (A, n = 10), chronic intermittent hypoxia group (B, n = 10), chronic intermittent hypoxia group (B, n = 10) and chronic intermittent hypoxia group (n = 2). Chronic intermittent hypoxia 2% ethanol melanin intraperitoneal injection group (C 10). The levels of superoxide dismutase (SOD),) malondialdehyde (MDA) and apoptosis index (bax,bcl-2 protein) in hippocampus were detected by chemical colorimetry and immunohistochemistry (SP method) respectively. The apoptosis rate of hippocampal neurons was detected by TUNEL method. Results the level of MDA [(0.6760 卤0.0687) nmol/mg protein] in hippocampus in MT group was significantly lower than that in normal control group [(1.1 卤0.16) nmol/mg protein] and CIH group [(1.7 卤0.17) nmol/mg protein]. The level of MDA in hippocampal tissue of rats in each group was significantly higher than that in the control group (F = 12.876, P < 0.05). The activity of SOD in hippocampus of P0.01). MT group [(46.92 卤5.66) U/mg protein] was significantly higher than that of CIH group ([32.288 卤2.984) U/mg protein], but lower than that of normal control group [(64.28 卤5.29) U/mg protein]. There were statistical differences among the three groups (F = 11.2525, F = 11.2525). Compared with CIH group (0.0968 卤0.0173) and normal group (0.142 卤0.0178), the gray value of bcl-2 protein positive cells in P0.01). MT treatment group (0.220 卤0.029) was significantly higher than that in CIH group (0.142 卤0.0178). The expression of bax protein in MT group (0.135 卤0.00688) was significantly lower than that in CIH group (0.222 卤0.0256) and normal control group (0.176 卤0.012). The gray value of apoptotic cells in MT group (0.1930 卤0.0202) was significantly lower than that in CIH group (0.335 卤0.0532), but higher than that in normal control group (0.0675 卤0.0125). Conclusion melatonin has obvious inhibitory effect on apoptosis of hippocampal neurons in chronic intermittent hypoxia rats and antioxidant stress. The decrease of bcl-2 expression and the increase of bax expression may be one of the mechanisms of apoptosis.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R766.43
本文编号：2319929
[Abstract]:Objective OSAHS is caused by apnea or hypopnea caused by repeated complete or partial obstruction of upper airway. The prominent pathophysiological process of OSAHS is repeated paroxysmal hypoxia / reoxygenation at night. Recurrent hypoxic / reoxygenation-chronic intermittent hypoxia can lead to multiple organ damage, especially in the nervous system, such as memory loss, cognitive impairment and so on. A large number of hippocampal neurons apoptosis is the key to learning and cognitive impairment in patients with OSAHS. Reactive oxygen species (Ros) play a key role in the long-term damage of hippocampal neurons caused by chronic intermittent hypoxia. Continuous positive airway pressure ventilation (CPAP) is the most effective method in the treatment of OSAHS, but there are still some patients who are unable to tolerate ventilator and give up treatment. Especially mild to moderate sleep apnea patients with poor compliance. Chronic intermittent hypoxia is the most important pathophysiological feature in patients with OSAHS. Therefore, the mechanism of chronic intermittent hypoxia on target organ damage and the corresponding intervention methods have become the focus of clinical research. Therefore, we selected the chronic intermittent hypoxia rat model to study the pathophysiological process of hypoxia / reoxygenation in patients with OSAHS, and treated with antioxidant melatonin, and observed the oxidative stress index MDA, in the hippocampus of chronic intermittent hypoxia. Expression of SOD and bcl-2,bax protein and apoptosis of hippocampal neurons. To investigate the therapeutic effect of antioxidant MT in CIH rat model and its related mechanism. Methods Thirty 2-month-old SD healthy male rats were randomly divided into 2% normoxic saline group (A, n = 10), chronic intermittent hypoxia group (B, n = 10), chronic intermittent hypoxia group (B, n = 10) and chronic intermittent hypoxia group (n = 2). Chronic intermittent hypoxia 2% ethanol melanin intraperitoneal injection group (C 10). The levels of superoxide dismutase (SOD),) malondialdehyde (MDA) and apoptosis index (bax,bcl-2 protein) in hippocampus were detected by chemical colorimetry and immunohistochemistry (SP method) respectively. The apoptosis rate of hippocampal neurons was detected by TUNEL method. Results the level of MDA [(0.6760 卤0.0687) nmol/mg protein] in hippocampus in MT group was significantly lower than that in normal control group [(1.1 卤0.16) nmol/mg protein] and CIH group [(1.7 卤0.17) nmol/mg protein]. The level of MDA in hippocampal tissue of rats in each group was significantly higher than that in the control group (F = 12.876, P < 0.05). The activity of SOD in hippocampus of P0.01). MT group [(46.92 卤5.66) U/mg protein] was significantly higher than that of CIH group ([32.288 卤2.984) U/mg protein], but lower than that of normal control group [(64.28 卤5.29) U/mg protein]. There were statistical differences among the three groups (F = 11.2525, F = 11.2525). Compared with CIH group (0.0968 卤0.0173) and normal group (0.142 卤0.0178), the gray value of bcl-2 protein positive cells in P0.01). MT treatment group (0.220 卤0.029) was significantly higher than that in CIH group (0.142 卤0.0178). The expression of bax protein in MT group (0.135 卤0.00688) was significantly lower than that in CIH group (0.222 卤0.0256) and normal control group (0.176 卤0.012). The gray value of apoptotic cells in MT group (0.1930 卤0.0202) was significantly lower than that in CIH group (0.335 卤0.0532), but higher than that in normal control group (0.0675 卤0.0125). Conclusion melatonin has obvious inhibitory effect on apoptosis of hippocampal neurons in chronic intermittent hypoxia rats and antioxidant stress. The decrease of bcl-2 expression and the increase of bax expression may be one of the mechanisms of apoptosis.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R766.43
【参考文献】
相关期刊论文 前1条
1 刘辉国;张志锋;张珍祥;高永平;牛汝楫;;间歇缺氧大鼠海马神经元凋亡及其机制[J];中华结核和呼吸杂志;2007年05期
，本文编号：2319929
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