CAJ | 학술논문

目的探讨肝X受体(LXR)信号通路在内皮素-1(ET-1)诱导的小鼠HL-1心肌细胞肥大中的作用.方法将培养的小鼠HL-1心肌细胞分为4组:(1)对照组:加入DMSO；(2)T0901317组:加入LXR受体激动剂T0901317(终浓度为1μmol/L)；(3)ET-1组:加入ET-1(终浓度1 nmol/L)诱导心肌细胞肥大；(4)T0901317+ ET-1组:先加入T0901317(终浓度1μmol/L)处理8h后,再加入ET-1(终浓度1 nmol/L).采用免疫荧光技术进行细胞染色,NIH图像J处理软件分析细胞表面积,3H-亮氨酸的掺入检测心肌细胞蛋白合成速率,实时定量PCR法检测心钠肽(ANP)及β-肌球蛋白重链(β-MyHC) mRNA的表达水平,并观察沉默LXR表达后T0901317对ANP mRNA表达的影响.结果 ET-1组HL-1心肌细胞相对细胞表面积、ANP、β-MyHC mRNA表达以及3H-亮氨酸的掺入分别为2.00±0.29、1.98±0.47、2.13 ±0.39和1.79 ±0.17,均显著高于对照组的1.00 ±0.26、1.00±0.21、1.00±0.31和1.00 ±0.03(P均＜0.05).T0901317+ ET-1组心肌细胞相对细胞表面积、ANP、β-MyHC mRNA表达及3H-亮氨酸的掺人分别为1.24±0.25、1.19 ±0.21、1.48±0.27和1.15 ±0.11,均显著低于ET-1组(P均＜0.05),沉默LXR α/β后T0901317+ET-1组ANP的mRNA表达为1.78±0.05,与ET-1组1.94 ±0.17比较差异无统计学意义.结论 LXR激动剂T0901317可抑制ET-1诱导的HL-1细胞的肥大,T0901317对ANP mRNA表达的抑制是受体依赖性的.
목적 탐토간X수체(LXR)신호통로재내피소-1(ET-1)유도적소서HL-1심기세포비대중적작용.방법 장배양적소서HL-1심기세포분위4조:(1)대조조:가입DMSO；(2)T0901317조:가입LXR수체격동제T0901317(종농도위1μmol/L)；(3)ET-1조:가입ET-1(종농도1 nmol/L)유도심기세포비대；(4)T0901317+ ET-1조:선가입T0901317(종농도1μmol/L)처리8h후,재가입ET-1(종농도1 nmol/L).채용면역형광기술진행세포염색,NIH도상J처리연건분석세포표면적,3H-량안산적참입검측심기세포단백합성속솔,실시정량PCR법검측심납태(ANP)급β-기구단백중련(β-MyHC) mRNA적표체수평,병관찰침묵LXR표체후T0901317대ANP mRNA표체적영향.결과 ET-1조HL-1심기세포상대세포표면적、ANP、β-MyHC mRNA표체이급3H-량안산적참입분별위2.00±0.29、1.98±0.47、2.13 ±0.39화1.79 ±0.17,균현저고우대조조적1.00 ±0.26、1.00±0.21、1.00±0.31화1.00 ±0.03(P균＜0.05).T0901317+ ET-1조심기세포상대세포표면적、ANP、β-MyHC mRNA표체급3H-량안산적참인분별위1.24±0.25、1.19 ±0.21、1.48±0.27화1.15 ±0.11,균현저저우ET-1조(P균＜0.05),침묵LXR α/β후T0901317+ET-1조ANP적mRNA표체위1.78±0.05,여ET-1조1.94 ±0.17비교차이무통계학의의.결론 LXR격동제T0901317가억제ET-1유도적HL-1세포적비대,T0901317대ANP mRNA표체적억제시수체의뢰성적.
Objective To investigate the role of liver X receptors (LXRs) on endothelin-1 ( ET-1 )induced murine HL-1 cardiomyocytes hypertrophy.Methods Cutured murine HL-1 cardiomyocytes were divided into four experiment groups:①Control group:treated with DMSO ; ②T0901317 group:treated with LXRs agonist T0901317( 1 μmol/L) ; ③ET-1 group:treated with ET-1 ( 1 nmoL/L) ; ④T0901317 + ET-1 group:treated with T0901317 ( 1 μmol/L) for 8 hours,then treated with ET-1 ( 1 nmol/L).Twenty-four hours later,immunofluorescent staining was performed on HL-1 cells,the surface area of HL-1 cells was analyzed with NIH Image J software,and the synthetic rate of protein in HL-1 cells was detected by 3 H-leucine incorporation.The mRNA level of atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MyHC ) was measured by quantitative realtime PCR.The effect of T0901317 on mRNA expression of ANP was also detected after LXRs gene silencing.Results The surface area of HL-1 cells,mRNA expression of ANP and β-MyHC,and 3H-leucine incorporation in ET-1 group were 2.00 ± 0.29,1.98 ± 0.47,2.13 ±0.39 and 1.79 ±0.17,respectively,which were singnficantly higher than those of control group ( 1.00 ±0.26,1.00 ±0.21,1.00 ±0.31 and 1.00 ±0.03,respectively,all P ＜0.05).Compared with ET-1 group,the surface area of HL-1 cells,mRNA expression of ANP and β-MyHC,and 3H-leucine incorporation were significantly decreased in T0901317 + ET-1 group ( 1.24 ± 0.25,1.19 ± 0.21,1.48 ± 0.27 and 1.15 ±0.11,respectively,all P ＜ 0.05 ).After inhibition of LXRo/β expression in HL-1 cardiomyocytes using the specific siRNAs,the mRNA expression of ANP in T0901317 + ET-1 group was 1.78 ± 0.05,which was similar as that in ET-1 group ( 1.94 ± 0.17,P ＞ 0.05).Conclusion T0901317,an agonist of LXRs,could inhibit ET-1 induced cardiac hypertrophy in vitro,and LXR Iigand-mediated inhibition on ANP mRNA expression by T0901317 is receptor dependent.