IR@PKUHSC  > 北京地坛医院教学医院
学科主题内科学
O-糖基化修饰抑制剂Benzyl-α-GalNAc对肝纤维化进程 的影响及可能的分子机制
张晓静
2014-05-20
导师魏红山
专业内科学
授予单位北京大学
授予地点北京地坛医院教学医院
学位硕士
关键词肝纤维化 O-糖基化修饰 Benzyl-α-GalNAc C5ORF55
其他题名The inhibitor of glycosylation resisits hapatic fibrosis and its possible mechanism
分类号R575.2
摘要目的:1)探讨O-糖基化抑制剂Benzyl-α-GalNAc对四氯化碳(CCl4)诱导小鼠肝纤维化进程的影响及部分可能的分子机制。2)克隆表达O-糖基化修饰相关分泌蛋白C5ORF55。方法:(1)动物分组:将雌性BALB/C小鼠120只(8-10周)随机均等分为,正常对照组(Control group, C);CCl4模型组(Model group, M);Benzyl-α-GalNAc低剂量组(Low Drug group, DL);Benzyl-α-GalNAc高剂量组(High Drug group,DH)。除C组外,M,DL,DH组小鼠腹腔注射50%CCl4(CCl4:橄榄油=1:1,1 ml/g,每周两次),DH组给Benzyl-α-GalNAc灌胃(5 mg/kg,1 次/天),DL组给Benzyl-α-GalNAc灌胃(1 mg/kg,1 次/天)。分别于第2周末,第4周末,第6周末处死各组小鼠10只,HE及Masson染色观察肝组织的病理学改变;分离血清检测肝功能指标;Western blot方法检测肝组织表达的转化生长因子β法受体(TβR1)、α-平滑肌肌动蛋白(α-SMA)、内质网应激标志物GRP78变化。(2)油红O染色,观察Benzyl-α-GalNAc对HepG2、L02脂质代谢的影响。(3)克隆表达O-糖基化修饰相关分泌蛋白C5ORF55的表达。结果:(1)第2周的血清ALT及AST治疗组同模型组相比有统计学差异(P<0.05),第4周及第6周血清学治疗组同模型组相比无显著统计学差异;第2周肝组织病理HE及Masson染色显示治疗组同模型组相比有统计学差异(P<0.05),第4周及第6周肝组织病理HE及Masson染色显示治疗组同模型组相比无显著统计学差异;第2周肝组织表达蛋白分子TβR1、α-SMA及GRP78治疗组同模型组相比有统计学差异(P<0.05),第4周及第6周肝组织表达蛋白分子TβR1、α-SMA及GRP78治疗组同模型组相比无显著统计学差异。(2)O-糖基化抑制剂Benzyl-α-GalNAc促进HepG2细胞脂肪变、及抑制其粘附,促进L02细胞脂肪变。(3)糖蛋白C5ORF55成功克隆表达及多抗的制备,且其只表达在肝组织、HepG2细胞及LX2细胞,在PBMC、淋巴结、胸腺中不表达。结论:(1)O-糖基化抑制剂Benzyl-α-GalNAc在一定剂量范围内抑制肝纤维化的形成和发展,高剂量可促进肝细胞脂肪变。提示,抑制Glcα1,2Galβ1-糖基化修饰,及/或其它类型的O-糖基化修饰,可能有助于延缓肝纤维化的进程。(2)糖蛋白C5ORF55主要表达于肝组织,肝实质细胞及/或其它肝细胞。
英文摘要Objective: (1) To explore the O-glycosylation inhibitor of Benzyl-α-GalNAc affects the influence of the process of mice liver fibrosis induced by carbon tetrachloride and the possible molecular mechanisms. (2) Clone and express O-glycosylation secretory protein C5ORF55. Methods: (1)Femal Balb/c mice (8-10 weeks old) were randomly divided into four groups, Control group, C; Model group, M; Low Drug group, DL; High Drug group,DH, each group has 30 mice. In addition to C group, M, DH, DL groups were intraperitoneally injected with equal amount and frequency CCl4/olive solution (1 ml/kg, twice per week). DH group were treated by the Benzyl-α-GalNAc once a day at 5 mg/kg; DL group were treated by the Benzyl-α-GalNAc once a day at 1 mg/kg. 10 mice of each group were killed after second, 4th and 6th weekend. The parameters were observed the changes of liver pathology by HE and Masson staining with microscopy, the serum alamine-aminotrasferase (ALT), aspartate-aminotrasferase (AST), total bilirubin (TBIL). Hepatic expression of TβR1, α-SMA and GRP78 were observed by western blot. (2) It was observed that Benzyl-α-GalNAc affects the cells of HepG2 and L02 by red oil O staining. (3) Clone and induce the expression of C5ORF55. Results: (1) The general conditions were better and the levels of ALT, AST were obvious in therapeutic groups compared with model groups (P<0.05) at the 2nd weekend, but there was no statistically significant difference at 4th and 6th weekend. The hepatic pathology indicated that HE and Masson staining has statistical difference(P<0.05) compared the groups of therapeutic groups to model groups at the 2nd weekend, but there was no statistically significant difference at 4th and 6th weekend. Liver tissue proteins TβR1, α-SMA and GRP78 have statistically significant difference (P<0.05), however, there was no significant difference between model groups and therapeutic groups at 4th, 6th weekend. (2) The inhibitor of O-glycosylation Benzyl-α–GalNAc promotes HepG2 cells fat and inhibits the cells adhesion, and promotes L02 cells fatty changes. The inhibitor of O-glycosylation Benzyl-α–GalNAc inhibits the adhesion of hepatic cells. (3) Glycoprotein C5ORF55 successfully was cloned and expressed, and only expressed in liver tissue, the LX2 cells and HepG2 cells, however, not expressed in PBMC, lymph nodes and thymus. Conclusions: (1) O-glycosylation inhibitor Benzyl-α–GalNAc may be slow the progression of liver fibrogenesis with a certain range dose, however, high dose can induce hepatocyte steatosis. (2) C5ORF55 staining mainly scattered in liver tissue, suggested that C5ORF55 may be secreted by hepatocytes and/or other liver cells.
语种中文
出处http://xuewei.bjmu.edu.cn/simpsearch.action?keyword=O-糖基化修饰抑制剂Benzyl-α-GalNAc对肝纤维化进程 的影响及可能的分子机制&dbid=72
文献类型学位论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/105800
专题北京地坛医院教学医院
作者单位北京地坛医院教学医院
推荐引用方式
GB/T 7714
张晓静. O-糖基化修饰抑制剂Benzyl-α-GalNAc对肝纤维化进程 的影响及可能的分子机制[D]. 北京地坛医院教学医院. 北京大学,2014.
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[张晓静]的文章
百度学术
百度学术中相似的文章
[张晓静]的文章
必应学术
必应学术中相似的文章
[张晓静]的文章
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。