衰老雄性大鼠脂质过氧化对血清睾酮水平和bcl-2、caspase-3基因表达的相关研究
发布时间：2018-11-03 14:56
【摘要】： 背景与目的近年来，大量的研究表明，在衰老进程中机体内氧化与抗氧化系统之间的平衡发生变化，，表现为氧化作用增强、抗氧化能力减弱，而细胞代谢异常产生大量活性氧ROS如超过抗氧化体系的还原能力，使细胞处于氧化应激状态，将影响到细胞功能状态，甚至引起凋亡相关基因表达的改变，激活细胞凋亡程序。为了减少有氧代谢过程中ROS对细胞的损害，细胞内有一系列有效的抗氧化防御机制，包括清除ROS的抗氧化酶如超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)等和阻断过氧化链式反应的生育酚、谷胱甘肽和抗坏血酸等。通常细胞处于氧化与抗氧化平衡中而维持着正常的功能，一旦氧化与抗氧化作用失衡，将会影响到细胞的功能。在衰老过程中，中老年男性出现雄激素部分缺乏，由此引起一系列的临床症状，但由于老年男性受到老龄合并疾病因素的影响，本实验拟建立D-半乳糖大鼠亚急性衰老模型研究脂质过氧化对血清睾酮水平、睾丸间质细胞bcl-2和caspase-3基因表达情况以及睾丸间质细胞的超微结构的影响，探讨衰老过程中脂质过氧化对血清睾酮水平的影响。 材料和方法 动物：SD大鼠20只，2～3月龄，体重(200±20)g，随机分成正常对照组、D-半乳糖组，每组10只。所有大鼠同室分笼饲养，自然光照，自由饮水进食，室温，相对湿度50～60％。正常对照组每日腹部皮下注射生理盐水(0.2ml／100g)6周；D-半乳糖组每日腹部皮下注射经高压灭菌D-半乳糖(200mg／kg)6周。于第6周末最后一次注射后48h，大鼠腹腔注射戊巴比妥钠(45mg／kg)麻醉，剪开阴囊，分离睾丸周围组织，结扎睾丸动脉，迅速取下左侧睾丸，称重，匀浆，分别检测组织中SOD、MDA、GSH-PX的水平，并取部分睾丸组织经2.5％戊二醛液固定，常规制作超薄片，用透射电镜观察超微结构；开胸暴露心脏，迅速从心尖部进针取血2ml，低温离心分离血清，置于4℃冰箱中保存，用于放射免疫法检测血清T；再剪开左心室插入灌注针头至升主动脉，用止血钳固定，剪开右心房，快速灌注生理盐水(室温)100～150ml，待血液冲洗干净后(肝脏变白)，即灌注4％多聚甲醛(4℃)500ml。取下右侧睾丸后，用4％多聚固定过夜，常规石蜡包埋、切片，检测睾丸间质bcl-2、caspase-3基因表达。 结果与正常对照组相比，D-半乳糖组Leydig细胞中内质网、线粒体数量减少；(1)SOD活性：D-半乳糖(D)组(116±18.09)NU／mgprot显著低于对照(C)组(156±31.02)NU／mgprot)(p＜0.01)；(2)MDA含量：D组(1.77±0.41)nmol／mgprot明显高于C组(1.19±0.15)nmol／mgprot(p＜0.05)；(3)GSH-PX水平：D组(29.84±3.79)U／mgprot显著降低于C组(35.29±4.50)U／mgprot(p＜0.05)；(4)血清T：D组(0.69±0.26)ng／ml显著低于C组(2.58±0.73)ng／ml(p＜0.01)；(5)bcl-2表达：D组(35.1±3.6)％明显低于C组(49.6±7.4)％(p＜0.01)；(6)caspase-3表达：D组(11.3±1.8)％明显高于C组(9.1±1.3)％(p＜0.01)。 结论衰老大鼠睾丸组织脂质过氧化的变化影响Leydig细胞超微结构、血清T水平和Leydig细胞bcl-2、caspase-3基因表达。
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, a large number of studies have shown that the balance between oxidation and anti-oxidation systems in the body changes during the aging process, which shows that oxidation is enhanced and anti-oxidation ability is weakened. and the cell metabolism abnormality generates a large amount of reactive oxygen ROS such as the reduction ability of the antioxidant system, so that the cell is in an oxidative stress state, the cell function state is influenced, and even the change of the expression of the apoptosis-related gene is caused, and the apoptosis program is activated. In order to reduce the damage of ROS to cells during aerobic metabolism, there are a series of effective anti-oxidant enzymes in cells, including antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), etc. which can scavenge ROS, and tocopherol which blocks the peroxidation chain reaction. glutathione and ascorbic acid, etc. Usually the cells are in the oxidation and antioxidant balance and maintain normal functions, which will affect the function of the cells once the oxidation and anti-oxidation effects are out-of-balance. In the aging process, the androgen part deficiency in middle-aged and old-aged men resulted in a series of clinical symptoms, but because older men were affected by the aging factors, The effects of lipid peroxidation on serum testosterone levels, the expression of bcl-2 and caspase-3 genes and the ultrastructure of testicular interstitial cells were studied in this study. Materials and Methods: 20 SD rats, 2 ~ 3 months old, weighing (200 ~ 20) g, randomly divided into normal pairs Group D-galactose group, 10 rats in each group. All rats were fed with cage, natural lighting and free drinking water. Food, room temperature, relative humidity 50-60%. Normal control group subcutaneous injection of normal saline (0.2ml/ 100g) for 6 weeks; D-galactose group daily abdominal subcutaneous injection through high-pressure sterilization D-galactose (200mg/ kg) for 6 weeks. After the last injection of the 6th week, 48h after the last injection of the rats, rats were anesthetized by intraperitoneal injection of Pentosamine (45mg/ kg), the scrotum was cut off, the surrounding tissues of the testis were separated, the testis artery was ligated, the left testis of the left side was rapidly removed, weighed and homogenized to detect the SO in the tissues respectively. D, MDA, GSH-PX levels were measured, and some testis tissues were fixed by 2.5% glutaraldehyde solution, and ultra-thin slices were fabricated by using transmission electron microscope, the ultrastructure was observed by transmission electron microscope, the heart was exposed to the open chest, the blood was rapidly extracted from the apical part into the needle, and the serum was centrifuged at low temperature. The serum was placed in a refrigerator at 4 鈩
本文编号：2308124
[Abstract]:BACKGROUND & OBJECTIVE: In recent years, a large number of studies have shown that the balance between oxidation and anti-oxidation systems in the body changes during the aging process, which shows that oxidation is enhanced and anti-oxidation ability is weakened. and the cell metabolism abnormality generates a large amount of reactive oxygen ROS such as the reduction ability of the antioxidant system, so that the cell is in an oxidative stress state, the cell function state is influenced, and even the change of the expression of the apoptosis-related gene is caused, and the apoptosis program is activated. In order to reduce the damage of ROS to cells during aerobic metabolism, there are a series of effective anti-oxidant enzymes in cells, including antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), etc. which can scavenge ROS, and tocopherol which blocks the peroxidation chain reaction. glutathione and ascorbic acid, etc. Usually the cells are in the oxidation and antioxidant balance and maintain normal functions, which will affect the function of the cells once the oxidation and anti-oxidation effects are out-of-balance. In the aging process, the androgen part deficiency in middle-aged and old-aged men resulted in a series of clinical symptoms, but because older men were affected by the aging factors, The effects of lipid peroxidation on serum testosterone levels, the expression of bcl-2 and caspase-3 genes and the ultrastructure of testicular interstitial cells were studied in this study. Materials and Methods: 20 SD rats, 2 ~ 3 months old, weighing (200 ~ 20) g, randomly divided into normal pairs Group D-galactose group, 10 rats in each group. All rats were fed with cage, natural lighting and free drinking water. Food, room temperature, relative humidity 50-60%. Normal control group subcutaneous injection of normal saline (0.2ml/ 100g) for 6 weeks; D-galactose group daily abdominal subcutaneous injection through high-pressure sterilization D-galactose (200mg/ kg) for 6 weeks. After the last injection of the 6th week, 48h after the last injection of the rats, rats were anesthetized by intraperitoneal injection of Pentosamine (45mg/ kg), the scrotum was cut off, the surrounding tissues of the testis were separated, the testis artery was ligated, the left testis of the left side was rapidly removed, weighed and homogenized to detect the SO in the tissues respectively. D, MDA, GSH-PX levels were measured, and some testis tissues were fixed by 2.5% glutaraldehyde solution, and ultra-thin slices were fabricated by using transmission electron microscope, the ultrastructure was observed by transmission electron microscope, the heart was exposed to the open chest, the blood was rapidly extracted from the apical part into the needle, and the serum was centrifuged at low temperature. The serum was placed in a refrigerator at 4 鈩
本文编号：2308124
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