中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
22期
10118-10123
,共6页
肖元元%徐淼%刘若冰%魏美林%韩峻峰%殷峻%黄金伟%魏丽
肖元元%徐淼%劉若冰%魏美林%韓峻峰%慇峻%黃金偉%魏麗
초원원%서묘%류약빙%위미림%한준봉%은준%황금위%위려
胰高血糖素样肽1%糖尿病%ob/ob小鼠%非酒精性脂肪肝
胰高血糖素樣肽1%糖尿病%ob/ob小鼠%非酒精性脂肪肝
이고혈당소양태1%당뇨병%ob/ob소서%비주정성지방간
Glucagon-like peptide 1%Diabetes mellitus%ob/ob mice%NAFLD
目的：观察胰高血糖素样肽1受体（GLP-1R）在ob/ob小鼠肝脏组织中的表达情况以及棕榈酸处理对肝癌细胞株 HepG2细胞 GLP-1R 的影响。方法（1）选择10周龄的雄性 ob/ob 小鼠（C57BL/6J-Lepob/J）与其同窝野生对照型小鼠各5只，取其肝脏组织，RT-PCR方法检测肝脏组织中GLP-1R基因的表达，Western blot法检测GLP-1R蛋白的表达水平，免疫组织化学染色法对肝脏组织进行染色观察。（2）不同浓度棕榈酸对待HepG2细胞24 h后，Western blot 法检测GLP-1R蛋白表达情况，RT-PCR法检测对细胞GLP-1R mRNA的影响。结果 GLP-1R在ob/ob小鼠肝脏中表达明显下降，蛋白表达水平分别为：0.7897±0.1458 vs.0.1361±0.0290，mRNA表达水分别为：0.8038±0.1222 vs.0.4234±0.1777，与对照组相比差异有统计学意义（P＜0.005）。随着棕榈酸浓度的升高，HepG2细胞GLP-1R水平在mRNA水平与蛋白水平均逐渐下降，且在200μmol/L时出现统计学差异。结论肝脏存在GLP-1R表达，脂肪的过度沉积可显著降低肝脏GLP-1R表达水平，从而影响肝脏对GLP-1的敏感性。
目的：觀察胰高血糖素樣肽1受體（GLP-1R）在ob/ob小鼠肝髒組織中的錶達情況以及棕櫚痠處理對肝癌細胞株 HepG2細胞 GLP-1R 的影響。方法（1）選擇10週齡的雄性 ob/ob 小鼠（C57BL/6J-Lepob/J）與其同窩野生對照型小鼠各5隻，取其肝髒組織，RT-PCR方法檢測肝髒組織中GLP-1R基因的錶達，Western blot法檢測GLP-1R蛋白的錶達水平，免疫組織化學染色法對肝髒組織進行染色觀察。（2）不同濃度棕櫚痠對待HepG2細胞24 h後，Western blot 法檢測GLP-1R蛋白錶達情況，RT-PCR法檢測對細胞GLP-1R mRNA的影響。結果 GLP-1R在ob/ob小鼠肝髒中錶達明顯下降，蛋白錶達水平分彆為：0.7897±0.1458 vs.0.1361±0.0290，mRNA錶達水分彆為：0.8038±0.1222 vs.0.4234±0.1777，與對照組相比差異有統計學意義（P＜0.005）。隨著棕櫚痠濃度的升高，HepG2細胞GLP-1R水平在mRNA水平與蛋白水平均逐漸下降，且在200μmol/L時齣現統計學差異。結論肝髒存在GLP-1R錶達，脂肪的過度沉積可顯著降低肝髒GLP-1R錶達水平，從而影響肝髒對GLP-1的敏感性。
목적：관찰이고혈당소양태1수체（GLP-1R）재ob/ob소서간장조직중적표체정황이급종려산처리대간암세포주 HepG2세포 GLP-1R 적영향。방법（1）선택10주령적웅성 ob/ob 소서（C57BL/6J-Lepob/J）여기동와야생대조형소서각5지，취기간장조직，RT-PCR방법검측간장조직중GLP-1R기인적표체，Western blot법검측GLP-1R단백적표체수평，면역조직화학염색법대간장조직진행염색관찰。（2）불동농도종려산대대HepG2세포24 h후，Western blot 법검측GLP-1R단백표체정황，RT-PCR법검측대세포GLP-1R mRNA적영향。결과 GLP-1R재ob/ob소서간장중표체명현하강，단백표체수평분별위：0.7897±0.1458 vs.0.1361±0.0290，mRNA표체수분별위：0.8038±0.1222 vs.0.4234±0.1777，여대조조상비차이유통계학의의（P＜0.005）。수착종려산농도적승고，HepG2세포GLP-1R수평재mRNA수평여단백수평균축점하강，차재200μmol/L시출현통계학차이。결론간장존재GLP-1R표체，지방적과도침적가현저강저간장GLP-1R표체수평，종이영향간장대GLP-1적민감성。
Objective To observe the expression of Glucagon-like peptide-1 receptor(GLP-1R) in the ob/ob mice liver and evaluate the effect of palmitic acid on GLP-1R in the HepG2 cell. Methods (1) we selected 5 10-week old male ob/ob mice and 5 their lean littermates. Collected their liver tissues, RT-PCR was used to detect GLP-1R mRNA expression in the liver, and the protein of GLP-1R was assayed by Western-blot and immunohistochemistry. (2) HepG2 cells were treated with different concentrations of palmitic acid, after 24 hours treatment, RT-PCR and Western blot were used to detect the change of mRNA and protein expression levels of GLP-1R. Results Compared with the littermates, the GLP-1R mRNA and protein in the liver tissues of ob/ob mice were significantly decreased (P<0.01), and the palmitic acid can reduce the mRNA and protein expression of GLP-1R in HepG2 cells, the effect was a dose-dependent manner(P<0.05). Conclusions GLP-1R is present in the liver, and the excessive fat deposition in the hepatocytes could decrease the expression of GLP-1R, which would affect the sensitivity of liver to the GLP-1.
