CAJ | 학술논문

目的探讨成纤维细胞生长因子21(FGF-21)基因表达下调对ApoE基因敲除(ApoE-/-)小鼠肝脏糖异生相关基因表达的影响及其可能的分子机制.方法 24只鼠龄8周的雄性ApoE-/-小鼠随机分为普食对照组(NC组,n=6),普食+pAd-shFGF-21组(NF组,n=6),高脂对照组(HC,n=6)和高脂+pAd-shFGF-21组(HF,n=6),各组小鼠均喂养16周.NC、HC组于15周末尾静脉注射Ad-shGFP空载腺病毒(浓度;1 ×109/100μl/鼠),NF、HF组注射pAd-shFGF-21重组腺病毒(浓度1×109/100μl/鼠).16周后取小鼠血浆和肝脏样本,测定空腹血糖、血浆胰岛素(PIns)和血脂水平；采用荧光定量PCR和Western印迹方法检测各组小鼠肝脏组织糖异生相关基因及JAK2/STAT3信号通路mRNA和蛋白水平的变化.结果 NF和HF组的空腹血糖、血浆胰岛素(PIns)、甘油三酯、总胆固醇及低密度脂蛋白胆固醇(LDL-C)水平均显著高于NC和HC组(P＜0.05或P＜0.01),而高密度脂蛋白胆固醇(HDL-C)水平显著下降(P＜0.05或P＜0.01).在普食与高脂饮食喂养条件下,FGFQ1抑制组与相应对照组相比肝脏FGF-21 mRNA水平分别降低76.6％和40.9％(均P＜0.05)；蛋白表达水平分别降低67.8％和49.6％(均P＜0.05)；β-klotho、FGFR1和FGFR3 mRNA水平显著降低(均P＜0.05),糖异生相关基因葡萄糖-6-磷酸酶(G6pase)和磷酸烯醇式丙酮酸羧激酶(PEPCK) mRNA和蛋白表达水平较对照组明显升高(P＜0.05),肝脏中SOCS3表达水平降低(P＜0.05)和磷酸化的STAT3、SOCS3蛋白表达量降低(P＜0.05).结论 FGF-21抑制ApoE-/-小鼠糖异生相关基因表达；其调控机制可能与JAK2/STAT3信号通路相关.
목적 탐토성섬유세포생장인자21(FGF-21)기인표체하조대ApoE기인고제(ApoE-/-)소서간장당이생상관기인표체적영향급기가능적분자궤제.방법 24지서령8주적웅성ApoE-/-소서수궤분위보식대조조(NC조,n=6),보식+pAd-shFGF-21조(NF조,n=6),고지대조조(HC,n=6)화고지+pAd-shFGF-21조(HF,n=6),각조소서균위양16주.NC、HC조우15주말미정맥주사Ad-shGFP공재선병독(농도;1 ×109/100μl/서),NF、HF조주사pAd-shFGF-21중조선병독(농도1×109/100μl/서).16주후취소서혈장화간장양본,측정공복혈당、혈장이도소(PIns)화혈지수평；채용형광정량PCR화Western인적방법검측각조소서간장조직당이생상관기인급JAK2/STAT3신호통로mRNA화단백수평적변화.결과 NF화HF조적공복혈당、혈장이도소(PIns)、감유삼지、총담고순급저밀도지단백담고순(LDL-C)수평균현저고우NC화HC조(P＜0.05혹P＜0.01),이고밀도지단백담고순(HDL-C)수평현저하강(P＜0.05혹P＜0.01).재보식여고지음식위양조건하,FGFQ1억제조여상응대조조상비간장FGF-21 mRNA수평분별강저76.6％화40.9％(균P＜0.05)；단백표체수평분별강저67.8％화49.6％(균P＜0.05)；β-klotho、FGFR1화FGFR3 mRNA수평현저강저(균P＜0.05),당이생상관기인포도당-6-린산매(G6pase)화린산희순식병동산최격매(PEPCK) mRNA화단백표체수평교대조조명현승고(P＜0.05),간장중SOCS3표체수평강저(P＜0.05)화린산화적STAT3、SOCS3단백표체량강저(P＜0.05).결론 FGF-21억제ApoE-/-소서당이생상관기인표체；기조공궤제가능여JAK2/STAT3신호통로상관.
Objective To investigate the effects of fibroblast growth factor-21 (FGF-21) gene knockdown on the expression of hepatic glyconeogenesis-related genes in ApoE-/-mice and its mechanism.Methods Twenty-four 8-week-old male ApoE-/-mice were randomly divided into chow diet fed group(NC,n =6),chow diet+pAd-shFGF-21group(NF,n=6),high-fat diet fed group(HC,n=6),and high-fat diet+pAd-shFGF-21 group(HF,n=6).Mice were fed for 16 weeks.NC and HC groups injected with pAd-shGFP,however NF and HF groups were injected with pAd-shFGF-21 through the tail vein at the end of the 15th week.At the end of the 16th week,blood samples were collected for measurement of plasma glucose,insulin and lipid levels.Hepatic tissues were collected for measurement of mRNA and protein levels of glyconeogenesis-related genes and JAK2/STAT3 signaling pathway via real-time PCR and Western-blot,respectively.Results The fasting plasma glucose (FBG),fasting plasma insulin (PIns),triglycerides,total cholesterol,and low-density lipoprotein-cholesterol (LDL-C) were significantly higher,while the high-density lipoprotein-cholesterol(HDL-C)was lower in FGF-21-knockdown groups compared with control groups,respectively(P＜0.05 or P＜0.01).When fed with normal and high-fat diet,hepatic FGF-21 mRNA levels were decreased by 76.6％ (P＜0.05) and 40.9％ (P＜0.05) compared with those in control groups,while hepatic FGF-21protein levels were decreased by 67.8％ (P＜0.05) and 49.6％ (P＜0.05),respectively.The hepatic glucose-6-phosphatase(G6pase) and phosphoenolpyruvate carboxykinase (PEPCK)mRNA and protein expression in FGF-21-knockdown groups were significantly increased compared with those in control groups(P＜0.05).At the same time we observed decreased expression of hepatic SOCS3 mRNA and decreased expression of phosphorylated STAT3 and SOCS3 protein in FGF-21 knockdown groups.Conclusions FGF-21 can repress the expression of liver glyconeogenesis-related genes and its mechanism might involve the JAK2/STAT3 signaling pathway.