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Mitochondrial DNA deletion and apoptosis in alcoholic liver disease

酒精性肝病中的线粒体 DNA 缺失和细胞凋亡

基本信息

批准号：
12670528
负责人：
SUCHISITIMA Mutsumi
金额：
$ 2.18万
依托单位：
KANAZAWA MEDICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670528/
关键词：
Alcoholic liver disease Mitochondoria DNA Apoptosis Necrosis ATP リポポリサッカライド Alcoholism DNA Mitochondria Mutations Deletions Apoptosis

项目摘要

Background : Chronic alcohol consumption depresses ATP synthesis and induces mitochondrial DNA (Mt-DNA) deletion in the liver. ATP content in cells may play a critical role in inducing cell death, apoptosis or necrosis. However, it is not known which type of cell death occurs in alcoholic liver disease. In this study, the deletions of hepatic Mt-DNA including ATPase (ATP synthase) encoding region and hepatic ATP content in rats treated with ethanol were determined to elucidate the relationship between Mt-DNA deletion and ATP synthesis. Single stranded DNA (ss-DNA) of hepatocytes was also stained immunohistochemically to determine the relationship between hepatic ATP content and apoptosis.Methods : Sixteen male Wistar rats were fed with a 36% ethanol-containing liquid diet (AL-group) or control diet without ethanol (C-group) for 5 weeks. Hepatic ATP content was measured in the extract of fresh liver tissue. To detect apoptosis of hepatocytes, ss-DNA in liver sections was stained immunoh … More istochemically. Ss-DNA positive cells per 10,000 hepatocytes of each liver tissue section were calculated. Using frozen liver sections, the deletions of Mt-DNA encoding complexes I (ND4 + ND5), IV (CO I, II and III) and V (ATPase 8/6) were determined by polymerase chain reaction (PCR).Results : Fatty change was observed in more than one third of lobules in AL-group, but not in C-group. Hepatic ATP content in AL-group was 0.44 ± 0.10 μmol/g liver which was significantly lower than that in C-group, 0.84 ± 0.31 μmol/g liver (P<0.005). On the other hand, no deletion of Mt-DNA encoding complexes I, IV and V was detected in either AL- or C-group. Ss-DNA staining was clearly observed in the nuclei of hepatocytes in both groups. The number of ss-DNA positive hepatocytes in AL-group was 5.6 ± 1.8/10,000 hepatocytes which was significantly less than that in C-group, 20.6 ± 4.8/10,000 hepatocytes (P<0.0001). There was a positive correlation between hepatic ATP content and the number of ss-DNA positive cells (r = 0.674, P<0.005).Conclusion : The results suggest that mitochondrial function, at least in part ATP level, was depressed before the damage of Mt-DNA by chronic ethanol consumption. Chronic ethanol consumption may not be responsible for the apoptosis of hepatocytes. Less

背景：长期饮酒抑制ATP合成，并诱导肝脏线粒体DNA（Mt-DNA）缺失。细胞内ATP含量在诱导细胞死亡、凋亡或坏死中起关键作用。然而，目前尚不清楚酒精性肝病中发生哪种类型的细胞死亡。本研究通过测定乙醇染毒大鼠肝脏线粒体DNA（Mt-DNA）包括ATP合成酶（ATP synthase，ATP synthase）编码区的缺失和肝脏ATP含量，探讨Mt-DNA缺失与ATP合成的关系。方法：雄性Wistar大鼠16只，以含36%乙醇的流质饲料（AL组）或不含乙醇的对照饲料（C组）喂养5周，取肝细胞进行ss-DNA染色，观察肝细胞ATP含量与细胞凋亡的关系。在新鲜肝组织提取物中测定肝ATP含量。为了检测肝细胞凋亡，对肝组织切片进行ss-DNA免疫染色， ...更多信息组织化学上计算每个肝组织切片的每10，000个肝细胞的Ss-DNA阳性细胞。采用冰冻切片，PCR检测编码复合物I（ND 4 + ND 5）、IV（CO I、II和III）和V（ATPase 8/6）的Mt-DNA的缺失。AL组肝组织ATP含量为0.44 ± 0.10 μmol/g肝，显著低于C组的0.84 ± 0.31 μmol/g肝（P<0.005）。另一方面，在AL-或C-组中均未检测到编码复合物I、IV和V的Mt-DNA的缺失。两组肝细胞核内均可见明显的Ss-DNA染色。AL组ss-DNA阳性肝细胞数为5.6 ± 1.8/10000个，明显低于C组的20.6 ± 4.8/10000个（P<0.0001）。肝线粒体ATP含量与ss-DNA阳性细胞数呈正相关（r = 0.674，P<0.005）。结论：慢性酒精性肝损伤前线粒体功能，至少部分ATP水平受到抑制。慢性乙醇消耗可能不是肝细胞凋亡的原因。少