中华实验和临床感染病杂志(电子版)
中華實驗和臨床感染病雜誌(電子版)
중화실험화림상감염병잡지(전자판)
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL INFECTIOUS DISEASES(ELECTRONIC VERSION)
2014年
1期
7-11
,共5页
王晶晶%刘顺爱%张锦前%全敏%冯胜虎%王琦%赵龙凤%成军
王晶晶%劉順愛%張錦前%全敏%馮勝虎%王琦%趙龍鳳%成軍
왕정정%류순애%장금전%전민%풍성호%왕기%조룡봉%성군
肝炎病毒,丙型%核心蛋白%肝X受体α%脂肪变
肝炎病毒,丙型%覈心蛋白%肝X受體α%脂肪變
간염병독,병형%핵심단백%간X수체α%지방변
Heptitis C virus%Core%Liver X receptor (LXRα)%Steatosis
目的:观察丙型肝炎病毒(HCV)的核心蛋白(core)对肝细胞甘油三酯(TG)合成的影响。方法将HCV 1b基因型core表达载体pcDNA3.1-myc/his(-)-core1b和HCV 3a基因型core表达载体pcDNA3.1-myc/his(-)-core3a分别转染至HepG2细胞,利用定量测定法检测细胞内TG含量,用实时荧光定量逆转录聚合酶链反应法(real time qPCR)、蛋白免疫印迹(Western blot)方法观察脂质合成相关基因肝X受体α(LXRα)、甾醇调节元件结合蛋白1c(SREBP1c)和脂肪酸合酶(FASN)的mRNA和蛋白的表达变化,并进行比较分析。结果 HepG2细胞转染两个基因型HCV core表达载体之后均能显著增加细胞内的TG含量(P<0.05),尤以基因型core3a组为差异更显著(P<0.001)。Real-time PCR结果显示,两个基因型core均上调LXRα的mRNA水平(P<0.05),基因型core3a组差异更为显著(P<0.01);FASN的mRNA水平只在仅在基因型core3a组上调(P<0.05),而基因型core1b组差异无统计学意义。Western blot结果显示,HCV core表达可以明显上调表达可以显著上调SREBP1c的蛋白水平,尤其基因型core3a组更为显著。对FASN蛋白水平,基因型core3a上调其表达,但基因型core1b组无明显变化。结论 HCV可能通过LXRα介导肝细胞内的脂质合成诱导肝脂肪变,有待进一步研究。
目的:觀察丙型肝炎病毒(HCV)的覈心蛋白(core)對肝細胞甘油三酯(TG)閤成的影響。方法將HCV 1b基因型core錶達載體pcDNA3.1-myc/his(-)-core1b和HCV 3a基因型core錶達載體pcDNA3.1-myc/his(-)-core3a分彆轉染至HepG2細胞,利用定量測定法檢測細胞內TG含量,用實時熒光定量逆轉錄聚閤酶鏈反應法(real time qPCR)、蛋白免疫印跡(Western blot)方法觀察脂質閤成相關基因肝X受體α(LXRα)、甾醇調節元件結閤蛋白1c(SREBP1c)和脂肪痠閤酶(FASN)的mRNA和蛋白的錶達變化,併進行比較分析。結果 HepG2細胞轉染兩箇基因型HCV core錶達載體之後均能顯著增加細胞內的TG含量(P<0.05),尤以基因型core3a組為差異更顯著(P<0.001)。Real-time PCR結果顯示,兩箇基因型core均上調LXRα的mRNA水平(P<0.05),基因型core3a組差異更為顯著(P<0.01);FASN的mRNA水平隻在僅在基因型core3a組上調(P<0.05),而基因型core1b組差異無統計學意義。Western blot結果顯示,HCV core錶達可以明顯上調錶達可以顯著上調SREBP1c的蛋白水平,尤其基因型core3a組更為顯著。對FASN蛋白水平,基因型core3a上調其錶達,但基因型core1b組無明顯變化。結論 HCV可能通過LXRα介導肝細胞內的脂質閤成誘導肝脂肪變,有待進一步研究。
목적:관찰병형간염병독(HCV)적핵심단백(core)대간세포감유삼지(TG)합성적영향。방법장HCV 1b기인형core표체재체pcDNA3.1-myc/his(-)-core1b화HCV 3a기인형core표체재체pcDNA3.1-myc/his(-)-core3a분별전염지HepG2세포,이용정량측정법검측세포내TG함량,용실시형광정량역전록취합매련반응법(real time qPCR)、단백면역인적(Western blot)방법관찰지질합성상관기인간X수체α(LXRα)、치순조절원건결합단백1c(SREBP1c)화지방산합매(FASN)적mRNA화단백적표체변화,병진행비교분석。결과 HepG2세포전염량개기인형HCV core표체재체지후균능현저증가세포내적TG함량(P<0.05),우이기인형core3a조위차이경현저(P<0.001)。Real-time PCR결과현시,량개기인형core균상조LXRα적mRNA수평(P<0.05),기인형core3a조차이경위현저(P<0.01);FASN적mRNA수평지재부재기인형core3a조상조(P<0.05),이기인형core1b조차이무통계학의의。Western blot결과현시,HCV core표체가이명현상조표체가이현저상조SREBP1c적단백수평,우기기인형core3a조경위현저。대FASN단백수평,기인형core3a상조기표체,단기인형core1b조무명현변화。결론 HCV가능통과LXRα개도간세포내적지질합성유도간지방변,유대진일보연구。
Objective To investigate the effect of HCV core protein on the intracellular triglyceride accumulation in HepG2 cells. Methods HCV core genes were cloned and expressed. Intracellular triglyceride was assessed by quantiifcation with adipogenesis assay kit. The inlfuence of HCV core protein on the liver X receptor (LXRα), sterol regulatory element binding protein-1c (SREBP1c) and fatty acid synthase (FASN) mRNA expression were analyzed in vitro by real-time PCR. Increased protein expression of SREBP1c and FASN were observed by Western blot. Results There were signiifcant accumulation of triglycerides in both core1b and core3a-expressing HepG2 cells (P<0.05), while core3a protein expression induced signiifcant TG accumulation (P<0.001). LXRαmRNA expression were upregulated in both cells expressing of HCV core1b (P<0.05) and core3a (P<0.01). FASN mRNA level was increased in core3a-expreession HepG2 cells (P<0.05), however, SREBP1c mRNA level was elevated, but with no signiifcant difference. This was accompanied by an increase in protein levels of SREBP1 and FASN. Conclusion LXR alpha-mediated regulation of triglyceride signiifcant by HCV core protein in HepG2 cells.