8周游泳运动对非酒精性脂肪肝大鼠睾丸生殖细胞凋亡的影响

发布时间：2018-06-03 00:10

本文选题：运动 + 非酒精性脂肪肝　；参考：《成都体育学院》2016年硕士论文

【摘要】：目的:观察8周游泳运动对非酒精性脂肪肝(NAFLD)大鼠生殖细胞凋亡以及Fas/FasL、Caspase-3表达的影响,探讨有氧运动干预NAFLD大鼠生殖细胞凋亡的可能机制。方法:50只成年雄性SD大鼠随机分为对照组(C组,n=20)和模型组(M组,n=30)两组;C组再分为8周对照组(C8)和16周对照组(C16),M组又分为8周模型组(M8)、16周模型组(M16)及运动模型组(EM)。C组基础饲料饲养,M组给予高脂饲料饲养8周以建立NAFLD大鼠模型。8周后C8、M8两组禁食12小时处死,取肝组织HE染色光镜下行组织病理学观察,检测血TG、TC、AST、ALT含量,评定NAFLD大鼠模型成功率;第9周开始,EM组进行为期8周(60min/d,6d/w)无负重游泳训练。运动结束后处死C16、M16、EM三组大鼠,检测血TG、TC、AST、ALT含量,取肝组织、睾丸组织及睾周脂肪分别称重记录保存,HE染色光镜下观察肝组织、睾丸组织病理形态;采用原位末端转移酶标记法(TUNEL法)标记睾丸生殖细胞的凋亡情况;Western blot法检测各组大鼠睾丸组织Fas/FasL、Caspase-3蛋白表达情况。结果:(1)C8组未见明显肝细胞脂肪性变,M8组8只出现脂肪性变,M8组相对C8组肝细胞脂肪变程度积分显著增高;C16组2只可见肝细胞内少量脂肪性变,M16组全部大鼠脂肪性变明显,EM组中(n=9)6只可见脂肪性变;肝细胞脂肪变程度积分M16、EM组显著高于C16组,EM组显著低于M16组。(2)M8组血清AST、ALT、TC、TG较C8组显著增高;与C16组比较,M16组血清AST、ALT、TC、TG含量均显著增高,EM组TC含量显著增高,AST、ALT、TG含量虽有增高但组间比较无统计学差异。(3)M16和EM组睾周脂肪重量较C16组分别表现出轻度上升和下降,但组间差异无显著性;M16和EM组睾丸系数较C16组分别表现出轻度下降和上升,但无统计学差异,EM组大鼠睾丸系数较M16组显著上升。(4)M16组大鼠生精小管排列较疏松,生精细胞形态、排列异常;EM组大鼠生精小管较M16组排列紧密,生精细胞形态、排列未见明显异常。(5)与C16组比较,M16组睾丸生殖细胞凋亡明显增加,Fas/FasL、Caspase-3蛋白表达均显著上升,EM组凋亡程度增高,Fas/FasL、Caspase-3表达上调,但组间无显著差异;与M16组比较,EM组睾丸生殖细胞凋亡显著较少,Fas/FasL、Caspase-3蛋白表达均显著下降。结论:(1)高脂膳食可诱导SD大鼠NAFLD的形成,且随着高脂饲养时间的延长,NAFLD发生率上升,程度逐步加重。(2)NAFLD大鼠睾丸生殖细胞凋亡增多,而睾丸组织Fas/FasL、Caspase-3蛋白含量升高是引起凋亡增加的重要原因。(3)8周游泳运动明显减轻大鼠NAFLD程度,同时显著降低NAFLD大鼠睾丸生殖细胞凋亡程度,下调Fas/FasL、Caspase-3蛋白表达是其可能机制之一。
[Abstract]:Aim: to observe the effects of swimming for 8 weeks on the apoptosis of germ cells and the expression of Fas-FasL caspase-3 in rats with nonalcoholic fatty liver disease (NAF LDD), and to explore the possible mechanism of aerobic exercise on the apoptosis of germ cells in NAFLD rats. Methods Fifty adult male Sprague-Dawley rats were randomly divided into two groups: control group (C) and model group (n = 30). Group C was further divided into control group (n = 8) and control group (n = 16). Group M was fed with high fat diet for 8 weeks to establish NAFLD rat model. After 8 weeks, two groups were killed by fasting for 12 hours. The liver tissues were stained with HE staining to observe the histopathological changes, to detect the alt content of TGG TCCA, to evaluate the success rate of NAFLD rat model, and to conduct weightless swimming training in EM group for 8 weeks (60 min / d ~ 6 d / w) from the 9th week. At the end of exercise, three groups of rats were killed. The contents of alt in blood TGV TCU ASTT were measured. The liver tissue, testis tissue and peritestis fat were measured by HE staining light microscope to observe the histopathology of the liver and testis. Apoptosis of testicular germ cells was detected by in situ terminal transferase labeling (Tunel) and Western blot method was used to detect the expression of Fas-P / FasL- Caspase-3 protein in rat testis. Results there was no obvious fatty degeneration of hepatocytes in C8 group. The fatty degeneration in M8 group was significantly higher than that in C8 group. There were 2 rats in C16 group and all rats in M16 group were found to have adipose degeneration in liver cells. In EM group, 6 cases were found to have adipose degeneration. The liver cell fatty degree score in M16 EM group was significantly higher than that in C16 group, and was significantly lower in M16 group than that in M16 group. Compared with C16 group, the serum levels of TCTG in group M 16 were significantly higher than those in group C 16. The TC content in EM group was significantly higher than that in group C 16, but there was no significant difference between the two groups. The weight of peritestis fat in group EM and EM group showed a slight increase and decrease, respectively, compared with C16 group. However, there was no significant difference in testicular coefficient between M16 and EM groups compared with C16 group. However, there was no significant difference in testicular coefficient between EM group and M16 group. The testicular coefficient of EM group was significantly higher than that of M16 group. The arrangement of seminiferous tubules and the morphology of spermatogenic cells in EM group were looser than those in M16 group. The seminiferous tubules in EM group were arranged more closely than those in M16 group, and the morphology of spermatogenic cells in EM group was higher than that in M16 group. Compared with C16 group, the apoptosis of testicular germ cells in M16 group increased significantly, and the expression of Fas-Fas-Fas-Caspase-3 protein increased significantly in the EM group, but there was no significant difference between the two groups. Compared with M16 group, testicular germ cell apoptosis in EM group was significantly lower than that in M16 group. Conclusion: high fat diet can induce the formation of NAFLD in SD rats, and the incidence rate of NAFLD increases with the prolongation of high fat feeding time, and the degree of apoptosis of testicular germ cells increases gradually. The increase of caspase-3 protein content in testicular tissue is an important reason for the increase of apoptosis. Swimming for 8 weeks can significantly reduce the degree of NAFLD in rats, and decrease the degree of apoptosis of germ cells in testis of NAFLD rats. The down-regulation of Fas-FasL- Caspase-3 protein expression is one of the possible mechanisms.
【学位授予单位】：成都体育学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：G861.1;G804.2

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