中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2008年
12期
935-939
,共5页
脂肪肝%法尼醇%固醇调节元件结合蛋白1c%鹅去氧胆酸钠
脂肪肝%法尼醇%固醇調節元件結閤蛋白1c%鵝去氧膽痠鈉
지방간%법니순%고순조절원건결합단백1c%아거양담산납
Fatty liver%,Farnesol%Sterol regulatory element binding protein-lc%Sodium ehenodeoxycholate
目的 观察法尼醇X受体(FXR)对人肝细胞株L02细胞脂肪代谢的调节作用.方法 油酸钠建立L02细胞脂肪变模型(脂肪变组),油酸钠及鹅去氧胆酸钠建立干预模型(干预组),设对照组(培养基中不加药物),3组均设立24、48、72 h时间点.油红O染色,细胞内甘油三酯含量测定,检测肝细胞脂肪变程度;Westem blot和RT-PCR方法检测核受体FXR及固醇调节元件结合蛋白1c(SREBP-1c)的表达变化.结果 脂肪变组随着肝细胞脂肪变性的发生发展,FXR表达呈逐渐降低趋势,但差异无统计学意义.SREBP-lc mRNA的表达明显升高,24、48,72 h图像半定量分析结果分别为0.495±0.062、0.579±0.064、0.612±0.067,SREBP-1c蛋白表达分别为0.394±0.044、0.488±0.066、0.543±0.064,均明显高于干预组和对照组,差异有统计学意义.干预组FXR mRNA表达明显上调,24、48,72 h分别为0.253±0.041、0.298±0.042、0.334±0.051,FXR蛋白分别为0.221±-0.022、0.313±0.041,0.341±0.046,与脂肪变组比较,,值为6.41～50.93,Jp值均<0.05或0.01,差异有统计学意义.脂变组甘油三酯含量各时间段均明显增加,干预组甘油三酯含量各时间段均明显降低.差异有统计学意义.结论 FXR表达下调与肝细胞脂肪沉积密切相关;刺激FXR表达增加可能通过抑制其靶基因SREBP-1c表达而改善L02细胞脂肪变性.
目的 觀察法尼醇X受體(FXR)對人肝細胞株L02細胞脂肪代謝的調節作用.方法 油痠鈉建立L02細胞脂肪變模型(脂肪變組),油痠鈉及鵝去氧膽痠鈉建立榦預模型(榦預組),設對照組(培養基中不加藥物),3組均設立24、48、72 h時間點.油紅O染色,細胞內甘油三酯含量測定,檢測肝細胞脂肪變程度;Westem blot和RT-PCR方法檢測覈受體FXR及固醇調節元件結閤蛋白1c(SREBP-1c)的錶達變化.結果 脂肪變組隨著肝細胞脂肪變性的髮生髮展,FXR錶達呈逐漸降低趨勢,但差異無統計學意義.SREBP-lc mRNA的錶達明顯升高,24、48,72 h圖像半定量分析結果分彆為0.495±0.062、0.579±0.064、0.612±0.067,SREBP-1c蛋白錶達分彆為0.394±0.044、0.488±0.066、0.543±0.064,均明顯高于榦預組和對照組,差異有統計學意義.榦預組FXR mRNA錶達明顯上調,24、48,72 h分彆為0.253±0.041、0.298±0.042、0.334±0.051,FXR蛋白分彆為0.221±-0.022、0.313±0.041,0.341±0.046,與脂肪變組比較,,值為6.41～50.93,Jp值均<0.05或0.01,差異有統計學意義.脂變組甘油三酯含量各時間段均明顯增加,榦預組甘油三酯含量各時間段均明顯降低.差異有統計學意義.結論 FXR錶達下調與肝細胞脂肪沉積密切相關;刺激FXR錶達增加可能通過抑製其靶基因SREBP-1c錶達而改善L02細胞脂肪變性.
목적 관찰법니순X수체(FXR)대인간세포주L02세포지방대사적조절작용.방법 유산납건립L02세포지방변모형(지방변조),유산납급아거양담산납건립간예모형(간예조),설대조조(배양기중불가약물),3조균설립24、48、72 h시간점.유홍O염색,세포내감유삼지함량측정,검측간세포지방변정도;Westem blot화RT-PCR방법검측핵수체FXR급고순조절원건결합단백1c(SREBP-1c)적표체변화.결과 지방변조수착간세포지방변성적발생발전,FXR표체정축점강저추세,단차이무통계학의의.SREBP-lc mRNA적표체명현승고,24、48,72 h도상반정량분석결과분별위0.495±0.062、0.579±0.064、0.612±0.067,SREBP-1c단백표체분별위0.394±0.044、0.488±0.066、0.543±0.064,균명현고우간예조화대조조,차이유통계학의의.간예조FXR mRNA표체명현상조,24、48,72 h분별위0.253±0.041、0.298±0.042、0.334±0.051,FXR단백분별위0.221±-0.022、0.313±0.041,0.341±0.046,여지방변조비교,,치위6.41～50.93,Jp치균<0.05혹0.01,차이유통계학의의.지변조감유삼지함량각시간단균명현증가,간예조감유삼지함량각시간단균명현강저.차이유통계학의의.결론 FXR표체하조여간세포지방침적밀절상관;자격FXR표체증가가능통과억제기파기인SREBP-1c표체이개선L02세포지방변성.
Objective To investigate the effects of famesoid X receptor (FXR) on lipid metabolism in human hepatic L02 cells. Methods A steatosis model and an intervention model were established by treating human hepatocyte line L02 cells with sodium oleate or sodium oleate and sodium chenodeoxycholate (a natural agonist of FXR) respectively. Non-treated L02 cells served as controls. At three time points of 24, 48 and 72 hours, the accumulation of lipid droplets in the hepatocytes was observed by optical microscopy alter oil red O staining, and the the expression of FXR and SREBP- lc receptors was detected by RT-PCR and Western blot. Results Compared with the controls, expressions of FXR mRNA and protein were down-regulated gradually in the steatosis model at 24, 48 and 72 hours, FXR mRNA/beta-actin mRNA was 0.186± 0.02, 0.182 ± 0.028 and0.181 ± 0.022, FXRprotein/heta-tubulinproteinwas 0.105 5= 0.016,0.103±0.012 and 0.103±0.018, F from 0.01 to 0.14; 24 h vs 48 b, 48 vs72 h: P > 0.05. The expressions of SREBP-Ic mRNA and protein were increased gradually. At 24, 48 and 72 hours, SREBP-Ic mRNA/beta-aetin mRNA was 0.495±0.062, 0.579±0.064 and 0.612±0.067, SREBP-lc protein/beta-tubulin protein was 0.394± 0.044, 0.488 ± 0.066 and 0.543± 0.064, F from 0.80 to 4.66, 24 h vs 48 h,48 vs72 h: P < 0.05. In the intervention model, expressions of FXR mRNA and protein were increased markedly compared with the steatosis model. At 24, 48 and 72 hours, FXR mRNA/beta-aetin mRNA was 0.253 ± 0.041, 0.298 ± 0.042 and 0.334 ± 0.051, and FXR protein/beta-tubulin protein was 0.221 ± 0.022, 0.313 ± 0.041 and 0.341 ±0.046, F from 6.41 to 50.93, intervention models vs steatosis models at the same time points: P < 0.05-0.01.Expressions of SREBP-1 c mRNA and protem were significantly reduced. At 24, 48 and 72 hours, SREBP-1c mRNA/beta-actin mRNA was 0.296 ± 0.038, 0.328 ± 0.037 and 0.341 ± 0.055, and FXR protein/beta-tubulin protein was 0.295 ± 0.038, 0.334 ± 0.047 and 0.355 ± 0.054, F from 8.84 to 48.46; intervention models vs steatosis models at the same time point: P < 0.01. Both in the steatnsis model and the intervention model, content of TG and lipids accumulations were much more than those in the controls. Compared with the intervention model, levels of TG and lipids accumulation were markedly increased in the steatosis model at 24,48, 72 hours.At 24,48 and 72 honrs, TG/ceUular total protein in μg/mg was 173.0 ± 20.5,253.4 ± 36.1 and 361.2 ± 50.7 in the steatosis model, while in the intervention model the data was 84.1 ± 17.2, 113.0± 14.5 and 127.2 ± 20.1, F from 38.70 to 268.13, intervention models vs steatosis models at the same time point: P < 0.01. Conclusion Expression of FXR is closely associated with lipid homeostasis in hepatocytes. Up-regulation of the expression of FXR may improve lipidosis in L02 cells. Its possible mechanism involves reduction of SREBP-Ic expression and lipogenesis in hepatocytes.
