【摘要】：NAFLD是主要的現代文明病之一，嚴重危害人體健康。全球NAFLD的發(fā)病率約為20~30%，并且增長迅速，目前已成為全球最普遍的肝臟疾病。在NASH的形成過程中，肝細胞凋亡起著重要的作用，且肝細胞的凋亡水平和肝組織的炎癥程度有著密切的關系。所以，通過各種手段來降低肝細胞的凋亡水平，可以改善肝組織的炎癥反應，延緩或阻止NASH的進程。 研究目的：觀察耐力游泳運動對高脂膳食誘導大鼠NASH形成過程中大鼠血清FFA、TNF-α及肝組織凋亡相關蛋白的表達，探討運動在預防NASH形成過程中對肝細胞凋亡與肝細胞炎癥的影響規(guī)律，為運動預防和改善NASH提供一定的理論依據。 研究方法：Sprague-Dawley（SD）純系4周齡雄性大鼠84只，按膳食方式不同分為兩大組：普通膳食喂養(yǎng)組（普通組）和高脂膳食喂養(yǎng)組（高脂組）。兩組按是否有運動干預又分為安靜組和運動組兩個亞組，共計4個亞組，分別為普通膳食安靜組（C組）、普通膳食運動組（E組）、高脂膳食安靜組（H組）、高脂膳食運動組（HE組）。各組大鼠取材時間為實驗4周、8周和12周。制備肝組織常規(guī)石蠟切片進行肝細胞脂肪變性等級與炎癥程度評分；Western Blot方法測定肝臟Caspase-3、Bcl-2和Bax蛋白表達；流式細胞術檢測肝細胞凋亡率。 研究結果： （1）日平均攝入熱量：各實驗階段，日平均攝入熱量H組均高于C組（P＜0.01），E組均低于C組（P＜0.01），HE組均高于E組（P＜0.01），HE組均低于H組（P＜0.01）。 （2）體重與肝重：實驗4周和12周階段，體重H組均高于C組（P＜0.01）；實驗8周和12周時，E組均低于C組（P＜0.05），HE組均低于H組（P＜0.05）。各實驗階段大鼠肝重H組均高于C組（P＜0.01），HE組均高于E組（P＜0.01），且HE12組低于H12組（P＜0.05）。 （3）血清學指標：各實驗階段，血清FFA濃度H組均高于C組（P＜0.05），HE組均低于H組（P＜0.05）；實驗8周和12周階段，E組均低于C組（P＜0.01），且HE12組低于H12組（P＜0.01）。血清ALT僅H12組高于C12組（P＜0.01）。血清TNF-α水平結果為H12組高于C12組（P＜0.01），HE12組低于H12組（P＜0.05）。 （4）肝細胞凋亡率：各實驗階段，肝細胞凋亡率H組均高于C組（P＜0.05）；運動能使高脂膳食大鼠凋亡率下降，且HE12組低于H12組（P＜0.01）。 （5）肝組織凋亡相關蛋白：肝臟Caspase-3與Bax蛋白表達均表現為H8組高于C8組（P＜0.05）；H12組高于C12組（P＜0.01），HE12組高于E12組（P＜0.01），E12組低于C12組（P＜0.05），HE12組低于H12組（P＜0.01）。肝臟Bcl-2蛋白表達H4組低于C4組（P＜0.05），，HE4組低于E4組（P＜0.05）；H8組低于C8組（P＜0.01），HE8組低于H8組（P＜0.01）； H12組低于C12組（P＜0.01），E12組高于C12組（P＜0.05），HE12組高于H12組（P＜0.01），HE12組低于E12組（P＜0.01）。 （6）肝細胞脂肪變性與肝細胞炎癥活動度評分：肝細胞脂肪變性結果顯示，各實驗階段H組均高于C組（P＜0.01），HE組均高于E組（P＜0.01），H組均高于HE組（P＜0.01），其中H12組高于HE12組（P=0.068）。各實驗階段肝組織炎癥活動度評分C組均低于H組（P＜0.01），其中HE8組低于H8組（P=0.055），HE12組低于H12組（P＜0.01）。結論： 1．12周的高脂膳食可以成功復制大鼠NASH模型，其機制可能是高脂膳食導致了大鼠血清FFA增高引起肝細胞脂質化，并促進肝細胞凋亡和肝組織發(fā)生炎癥反應； 2．耐力游泳運動能有效緩解高脂膳食大鼠NASH的發(fā)展和改善NASH的癥狀，其機制可能是通過限制熱量的攝入、降低血FFA、減少肝細胞對脂質的積累，同時抑制肝細胞凋亡從而延緩NASH的進程并減輕肝組織炎癥活動； 3．在高脂膳食誘導的NASH形成過程中，肝細胞凋亡可能是促使肝臟單純脂肪變性發(fā)展成為NASH的一個重要原因。
[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.
【學位授予單位】：河北師范大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：G804.2

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