衰老雄性大鼠脂質(zhì)過(guò)氧化對(duì)血清睪酮水平和bcl-2、caspase-3基因表達(dá)的相關(guān)研究
發(fā)布時(shí)間：2018-11-03 14:56
【摘要】： 背景與目的近年來(lái)，大量的研究表明，在衰老進(jìn)程中機(jī)體內(nèi)氧化與抗氧化系統(tǒng)之間的平衡發(fā)生變化，，表現(xiàn)為氧化作用增強(qiáng)、抗氧化能力減弱，而細(xì)胞代謝異常產(chǎn)生大量活性氧ROS如超過(guò)抗氧化體系的還原能力，使細(xì)胞處于氧化應(yīng)激狀態(tài)，將影響到細(xì)胞功能狀態(tài)，甚至引起凋亡相關(guān)基因表達(dá)的改變，激活細(xì)胞凋亡程序。為了減少有氧代謝過(guò)程中ROS對(duì)細(xì)胞的損害，細(xì)胞內(nèi)有一系列有效的抗氧化防御機(jī)制，包括清除ROS的抗氧化酶如超氧化物歧化酶(SOD)、谷胱甘肽過(guò)氧化物酶(GSH-PX)等和阻斷過(guò)氧化鏈?zhǔn)椒磻?yīng)的生育酚、谷胱甘肽和抗壞血酸等。通常細(xì)胞處于氧化與抗氧化平衡中而維持著正常的功能，一旦氧化與抗氧化作用失衡，將會(huì)影響到細(xì)胞的功能。在衰老過(guò)程中，中老年男性出現(xiàn)雄激素部分缺乏，由此引起一系列的臨床癥狀，但由于老年男性受到老齡合并疾病因素的影響，本實(shí)驗(yàn)擬建立D-半乳糖大鼠亞急性衰老模型研究脂質(zhì)過(guò)氧化對(duì)血清睪酮水平、睪丸間質(zhì)細(xì)胞bcl-2和caspase-3基因表達(dá)情況以及睪丸間質(zhì)細(xì)胞的超微結(jié)構(gòu)的影響，探討衰老過(guò)程中脂質(zhì)過(guò)氧化對(duì)血清睪酮水平的影響。 材料和方法 動(dòng)物：SD大鼠20只，2～3月齡，體重(200±20)g，隨機(jī)分成正常對(duì)照組、D-半乳糖組，每組10只。所有大鼠同室分籠飼養(yǎng)，自然光照，自由飲水進(jìn)食，室溫，相對(duì)濕度50～60％。正常對(duì)照組每日腹部皮下注射生理鹽水(0.2ml／100g)6周；D-半乳糖組每日腹部皮下注射經(jīng)高壓滅菌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鈉(45mg／kg)麻醉，剪開陰囊，分離睪丸周圍組織，結(jié)扎睪丸動(dòng)脈，迅速取下左側(cè)睪丸，稱重，勻漿，分別檢測(cè)組織中SOD、MDA、GSH-PX的水平，并取部分睪丸組織經(jīng)2.5％戊二醛液固定，常規(guī)制作超薄片，用透射電鏡觀察超微結(jié)構(gòu)；開胸暴露心臟，迅速?gòu)男募獠窟M(jìn)針取血2ml，低溫離心分離血清，置于4℃冰箱中保存，用于放射免疫法檢測(cè)血清T；再剪開左心室插入灌注針頭至升主動(dòng)脈，用止血鉗固定，剪開右心房，快速灌注生理鹽水(室溫)100～150ml，待血液沖洗干凈后(肝臟變白)，即灌注4％多聚甲醛(4℃)500ml。取下右側(cè)睪丸后，用4％多聚固定過(guò)夜，常規(guī)石蠟包埋、切片，檢測(cè)睪丸間質(zhì)bcl-2、caspase-3基因表達(dá)。 結(jié)果與正常對(duì)照組相比，D-半乳糖組Leydig細(xì)胞中內(nèi)質(zhì)網(wǎng)、線粒體數(shù)量減少；(1)SOD活性：D-半乳糖(D)組(116±18.09)NU／mgprot顯著低于對(duì)照(C)組(156±31.02)NU／mgprot)(p＜0.01)；(2)MDA含量：D組(1.77±0.41)nmol／mgprot明顯高于C組(1.19±0.15)nmol／mgprot(p＜0.05)；(3)GSH-PX水平：D組(29.84±3.79)U／mgprot顯著降低于C組(35.29±4.50)U／mgprot(p＜0.05)；(4)血清T：D組(0.69±0.26)ng／ml顯著低于C組(2.58±0.73)ng／ml(p＜0.01)；(5)bcl-2表達(dá)：D組(35.1±3.6)％明顯低于C組(49.6±7.4)％(p＜0.01)；(6)caspase-3表達(dá)：D組(11.3±1.8)％明顯高于C組(9.1±1.3)％(p＜0.01)。 結(jié)論衰老大鼠睪丸組織脂質(zhì)過(guò)氧化的變化影響Leydig細(xì)胞超微結(jié)構(gòu)、血清T水平和Leydig細(xì)胞bcl-2、caspase-3基因表達(dá)。
[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
本文編號(hào)：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
本文編號(hào)：2308124
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