耐力游泳运动对高脂膳食诱导大鼠NASH形成过程中肝细胞凋亡的影响
发布时间：2018-09-07 15:19
【摘要】：NAFLD是主要的现代文明病之一，严重危害人体健康。全球NAFLD的发病率约为20~30%，并且增长迅速，目前已成为全球最普遍的肝脏疾病。在NASH的形成过程中，肝细胞凋亡起着重要的作用，且肝细胞的凋亡水平和肝组织的炎症程度有着密切的关系。所以，通过各种手段来降低肝细胞的凋亡水平，可以改善肝组织的炎症反应，延缓或阻止NASH的进程。 研究目的：观察耐力游泳运动对高脂膳食诱导大鼠NASH形成过程中大鼠血清FFA、TNF-α及肝组织凋亡相关蛋白的表达，探讨运动在预防NASH形成过程中对肝细胞凋亡与肝细胞炎症的影响规律，为运动预防和改善NASH提供一定的理论依据。 研究方法：Sprague-Dawley（SD）纯系4周龄雄性大鼠84只，按膳食方式不同分为两大组：普通膳食喂养组（普通组）和高脂膳食喂养组（高脂组）。两组按是否有运动干预又分为安静组和运动组两个亚组，共计4个亚组，分别为普通膳食安静组（C组）、普通膳食运动组（E组）、高脂膳食安静组（H组）、高脂膳食运动组（HE组）。各组大鼠取材时间为实验4周、8周和12周。制备肝组织常规石蜡切片进行肝细胞脂肪变性等级与炎症程度评分；Western Blot方法测定肝脏Caspase-3、Bcl-2和Bax蛋白表达；流式细胞术检测肝细胞凋亡率。 研究结果： （1）日平均摄入热量：各实验阶段，日平均摄入热量H组均高于C组（P＜0.01），E组均低于C组（P＜0.01），HE组均高于E组（P＜0.01），HE组均低于H组（P＜0.01）。 （2）体重与肝重：实验4周和12周阶段，体重H组均高于C组（P＜0.01）；实验8周和12周时，E组均低于C组（P＜0.05），HE组均低于H组（P＜0.05）。各实验阶段大鼠肝重H组均高于C组（P＜0.01），HE组均高于E组（P＜0.01），且HE12组低于H12组（P＜0.05）。 （3）血清学指标：各实验阶段，血清FFA浓度H组均高于C组（P＜0.05），HE组均低于H组（P＜0.05）；实验8周和12周阶段，E组均低于C组（P＜0.01），且HE12组低于H12组（P＜0.01）。血清ALT仅H12组高于C12组（P＜0.01）。血清TNF-α水平结果为H12组高于C12组（P＜0.01），HE12组低于H12组（P＜0.05）。 （4）肝细胞凋亡率：各实验阶段，肝细胞凋亡率H组均高于C组（P＜0.05）；运动能使高脂膳食大鼠凋亡率下降，且HE12组低于H12组（P＜0.01）。 （5）肝组织凋亡相关蛋白：肝脏Caspase-3与Bax蛋白表达均表现为H8组高于C8组（P＜0.05）；H12组高于C12组（P＜0.01），HE12组高于E12组（P＜0.01），E12组低于C12组（P＜0.05），HE12组低于H12组（P＜0.01）。肝脏Bcl-2蛋白表达H4组低于C4组（P＜0.05），，HE4组低于E4组（P＜0.05）；H8组低于C8组（P＜0.01），HE8组低于H8组（P＜0.01）； H12组低于C12组（P＜0.01），E12组高于C12组（P＜0.05），HE12组高于H12组（P＜0.01），HE12组低于E12组（P＜0.01）。 （6）肝细胞脂肪变性与肝细胞炎症活动度评分：肝细胞脂肪变性结果显示，各实验阶段H组均高于C组（P＜0.01），HE组均高于E组（P＜0.01），H组均高于HE组（P＜0.01），其中H12组高于HE12组（P=0.068）。各实验阶段肝组织炎症活动度评分C组均低于H组（P＜0.01），其中HE8组低于H8组（P=0.055），HE12组低于H12组（P＜0.01）。结论： 1．12周的高脂膳食可以成功复制大鼠NASH模型，其机制可能是高脂膳食导致了大鼠血清FFA增高引起肝细胞脂质化，并促进肝细胞凋亡和肝组织发生炎症反应； 2．耐力游泳运动能有效缓解高脂膳食大鼠NASH的发展和改善NASH的症状，其机制可能是通过限制热量的摄入、降低血FFA、减少肝细胞对脂质的积累，同时抑制肝细胞凋亡从而延缓NASH的进程并减轻肝组织炎症活动； 3．在高脂膳食诱导的NASH形成过程中，肝细胞凋亡可能是促使肝脏单纯脂肪变性发展成为NASH的一个重要原因。
[Abstract]:NAFLD is one of the major modern civilized diseases, which seriously endangers human health. The incidence of NAFLD is about 20-30% in the world, and it is growing rapidly. It has become the most common liver disease in the world. Hepatocyte apoptosis plays an important role in the formation of NASH, and the level of hepatocyte apoptosis is closely related to the degree of inflammation in liver tissue. Therefore, through various means to reduce the level of apoptosis of hepatocytes, can improve the inflammation of liver tissue, delay or prevent the process of NASH.
Objective: To observe the expression of FFA, TNF-a and apoptosis-related proteins in serum and liver tissues of rats during the process of NASH induced by high-fat diet during endurance swimming exercise, and to explore the effect of exercise on hepatocyte apoptosis and hepatocyte inflammation during the process of NASH prevention, so as to provide a theoretical basis for exercise prevention and improvement of NASH.
Methods: 84 Sprague-Dawley (SD) male rats aged 4 weeks were divided into two groups according to dietary patterns: normal diet group (normal group) and high-fat diet group (high-fat group). Normal dietary exercise group (E group), high-fat diet quiet group (H group), high-fat diet exercise group (HE group). The rats in each group were taken for 4 weeks, 8 weeks and 12 weeks. The apoptosis rate of hepatocytes was detected by cytometry.
Research findings:
(1) Daily average calorie intake: The daily average calorie intake of group H was higher than that of group C (P < 0.01), group E was lower than that of group C (P < 0.01), group HE was higher than that of group E (P < 0.01), and group HE was lower than that of group H (P < 0.01).
(2) Body weight and liver weight: At 4 and 12 weeks, the body weight of group H was higher than that of group C (P < 0.01); at 8 and 12 weeks, the body weight of group E was lower than that of group C (P < 0.05), and that of group HE was lower than that of group H (P < 0.05).
(3) Serum parameters: serum FFA concentration in H group was higher than that in C group (P < 0.05), and that in HE group was lower than that in H group (P < 0.05), and that in E group was lower than that in C group (P < 0.01) and HE12 group was lower than that in H12 group (P < 0.01). The group was lower than the H12 group (P < 0.05).
(4) Rate of hepatocyte apoptosis: The rate of hepatocyte apoptosis in group H was higher than that in group C (P < 0.05), and the rate of apoptosis in group HE12 was lower than that in group H12 (P < 0.01).
(5) Hepatic apoptosis-related proteins: the expression of Caspase-3 and Bax protein in H8 group was higher than that in C8 group (P < 0.05), H12 group was higher than that in C12 group (P < 0.01), HE12 group was higher than that in E12 group (P < 0.01), E12 group was lower than that in C12 group (P < 0.05), HE12 group was lower than that in H12 group (P < 0.01). The expression of Bcl-2 protein in H4 group was lower than that in C4 group (P < 0.05), and HE4 group was lower than that in E4 group (P < 0.05). H8 group was lower than C8 group (P < 0.01), HE8 group was lower than H8 group (P < 0.01), H12 group was lower than C12 group (P < 0.01), E12 group was higher than C12 group (P < 0.05), HE12 group was higher than H12 group (P < 0.01), HE12 group was lower than E12 group (P < 0.01).
(6) Hepatocyte steatosis and hepatocyte inflammatory activity score: The results of hepatocyte steatosis showed that H group was higher than C group (P < 0.01), HE group was higher than E group (P < 0.01), H group was higher than HE group (P < 0.01), and H12 group was higher than HE12 group (P = 0.068). 01), among which group HE8 was lower than group H8 (P=0.055), and group HE12 was lower than group H12 (P < 0.01).
1.12-week high-fat diet could successfully reproduce the NASH model of rats. The mechanism may be that high-fat diet could induce the increase of serum FFA, induce hepatocyte lipidization, and promote hepatocyte apoptosis and hepatic inflammation.
2. Endurance swimming can effectively alleviate the development of NASH and improve the symptoms of NASH in rats fed with high-fat diet. Its mechanism may be through limiting calorie intake, reducing blood FFA, reducing the accumulation of lipids in liver cells, and inhibiting hepatocyte apoptosis, thus delaying the process of NASH and alleviating inflammation in liver tissue.
3. Hepatocyte apoptosis may play an important role in the development of NASH induced by high-fat diet.
【学位授予单位】：河北师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：G804.2
本文编号：2228638
[Abstract]:NAFLD is one of the major modern civilized diseases, which seriously endangers human health. The incidence of NAFLD is about 20-30% in the world, and it is growing rapidly. It has become the most common liver disease in the world. Hepatocyte apoptosis plays an important role in the formation of NASH, and the level of hepatocyte apoptosis is closely related to the degree of inflammation in liver tissue. Therefore, through various means to reduce the level of apoptosis of hepatocytes, can improve the inflammation of liver tissue, delay or prevent the process of NASH.
Objective: To observe the expression of FFA, TNF-a and apoptosis-related proteins in serum and liver tissues of rats during the process of NASH induced by high-fat diet during endurance swimming exercise, and to explore the effect of exercise on hepatocyte apoptosis and hepatocyte inflammation during the process of NASH prevention, so as to provide a theoretical basis for exercise prevention and improvement of NASH.
Methods: 84 Sprague-Dawley (SD) male rats aged 4 weeks were divided into two groups according to dietary patterns: normal diet group (normal group) and high-fat diet group (high-fat group). Normal dietary exercise group (E group), high-fat diet quiet group (H group), high-fat diet exercise group (HE group). The rats in each group were taken for 4 weeks, 8 weeks and 12 weeks. The apoptosis rate of hepatocytes was detected by cytometry.
Research findings:
(1) Daily average calorie intake: The daily average calorie intake of group H was higher than that of group C (P < 0.01), group E was lower than that of group C (P < 0.01), group HE was higher than that of group E (P < 0.01), and group HE was lower than that of group H (P < 0.01).
(2) Body weight and liver weight: At 4 and 12 weeks, the body weight of group H was higher than that of group C (P < 0.01); at 8 and 12 weeks, the body weight of group E was lower than that of group C (P < 0.05), and that of group HE was lower than that of group H (P < 0.05).
(3) Serum parameters: serum FFA concentration in H group was higher than that in C group (P < 0.05), and that in HE group was lower than that in H group (P < 0.05), and that in E group was lower than that in C group (P < 0.01) and HE12 group was lower than that in H12 group (P < 0.01). The group was lower than the H12 group (P < 0.05).
(4) Rate of hepatocyte apoptosis: The rate of hepatocyte apoptosis in group H was higher than that in group C (P < 0.05), and the rate of apoptosis in group HE12 was lower than that in group H12 (P < 0.01).
(5) Hepatic apoptosis-related proteins: the expression of Caspase-3 and Bax protein in H8 group was higher than that in C8 group (P < 0.05), H12 group was higher than that in C12 group (P < 0.01), HE12 group was higher than that in E12 group (P < 0.01), E12 group was lower than that in C12 group (P < 0.05), HE12 group was lower than that in H12 group (P < 0.01). The expression of Bcl-2 protein in H4 group was lower than that in C4 group (P < 0.05), and HE4 group was lower than that in E4 group (P < 0.05). H8 group was lower than C8 group (P < 0.01), HE8 group was lower than H8 group (P < 0.01), H12 group was lower than C12 group (P < 0.01), E12 group was higher than C12 group (P < 0.05), HE12 group was higher than H12 group (P < 0.01), HE12 group was lower than E12 group (P < 0.01).
(6) Hepatocyte steatosis and hepatocyte inflammatory activity score: The results of hepatocyte steatosis showed that H group was higher than C group (P < 0.01), HE group was higher than E group (P < 0.01), H group was higher than HE group (P < 0.01), and H12 group was higher than HE12 group (P = 0.068). 01), among which group HE8 was lower than group H8 (P=0.055), and group HE12 was lower than group H12 (P < 0.01).
1.12-week high-fat diet could successfully reproduce the NASH model of rats. The mechanism may be that high-fat diet could induce the increase of serum FFA, induce hepatocyte lipidization, and promote hepatocyte apoptosis and hepatic inflammation.
2. Endurance swimming can effectively alleviate the development of NASH and improve the symptoms of NASH in rats fed with high-fat diet. Its mechanism may be through limiting calorie intake, reducing blood FFA, reducing the accumulation of lipids in liver cells, and inhibiting hepatocyte apoptosis, thus delaying the process of NASH and alleviating inflammation in liver tissue.
3. Hepatocyte apoptosis may play an important role in the development of NASH induced by high-fat diet.
【学位授予单位】：河北师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：G804.2
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