中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2009年
10期
776-780
,共5页
李树颖%陈睿%李晶%王宝利%于德民
李樹穎%陳睿%李晶%王寶利%于德民
리수영%진예%리정%왕보리%우덕민
脂肪肝%糖尿病%核糖体蛋白S6激酶1%固醇调节元件结合蛋白1c
脂肪肝%糖尿病%覈糖體蛋白S6激酶1%固醇調節元件結閤蛋白1c
지방간%당뇨병%핵당체단백S6격매1%고순조절원건결합단백1c
Fatty liver%Diabetes mellitus%Ribosomal protein S6 kinase 1%Sterol regulatory element binding protein 1c
目的 探讨核糖体蛋白S6激酶1(S6K1)基因沉默后对高糖刺激下小鼠肝细胞固醇调节元件结合蛋白1c(SREBP1c)表达的影响.方法 将构建的S6K1shRNA重组基因腺病毒(S6K1Ax)注射进db/db小鼠尾静脉,以注射含pU6启动子的腺病毒(pU6Ax)空载体的db/db小鼠作对照组,HE染色观察肝脏病理改变并检测肝脏甘油三酯含量变化.S6K1Ax转染小鼠肝细胞AML12细胞,转染pU6Ax的作为对照组,分别用高糖、高胰岛素,高糖高胰岛素刺激,逆转录聚合酶链式反应分析肝细胞在各种条件刺激后mSREBP1c等的表达,Western blot检测db/db小鼠肝脏及AML12细胞S6K1的蛋白表达.结果 db/db糖尿病小鼠注射S6K1Ax后1周,肝脏S6K1蛋白表达被抑制,对照组与实验组肝脏甘油三酯含量分别为(0.65±0.02)mmol/L和(0.56±0.01)mmol/L,t=4.312,P<0.01,差异有统计学意义.HE染色显示实验组肝细胞胞质含脂肪滴减少,空泡细胞数量减少,脂肪肝得到改善.AML12肝细胞mSREBP1c表达实验组为0.03±0.01,对照组为0.06±0.01,t=5.624,P<0.01,差异有统计学意义.与基础状态相比,给以胰岛素刺激后实验组和对照组mSREBP1c表达均增加,实验组上升至0.06±0.02,t=8.452,P<0.01,对照组上升至0.08±0.02,t=3.591,P<0.05.高糖刺激对实验组和对照组mSREBP1c表达均无明显影响.与高胰岛素刺激组相比,高糖高胰岛素刺激后实验组和对照组mSREBP1c表达均无明显差异.结论 S6K1通过影响脂代谢的关键调控基因mSREBP1c表达参与了脂肪的合成,推测S6K1在脂肪肝的形成中发挥了重要作用.
目的 探討覈糖體蛋白S6激酶1(S6K1)基因沉默後對高糖刺激下小鼠肝細胞固醇調節元件結閤蛋白1c(SREBP1c)錶達的影響.方法 將構建的S6K1shRNA重組基因腺病毒(S6K1Ax)註射進db/db小鼠尾靜脈,以註射含pU6啟動子的腺病毒(pU6Ax)空載體的db/db小鼠作對照組,HE染色觀察肝髒病理改變併檢測肝髒甘油三酯含量變化.S6K1Ax轉染小鼠肝細胞AML12細胞,轉染pU6Ax的作為對照組,分彆用高糖、高胰島素,高糖高胰島素刺激,逆轉錄聚閤酶鏈式反應分析肝細胞在各種條件刺激後mSREBP1c等的錶達,Western blot檢測db/db小鼠肝髒及AML12細胞S6K1的蛋白錶達.結果 db/db糖尿病小鼠註射S6K1Ax後1週,肝髒S6K1蛋白錶達被抑製,對照組與實驗組肝髒甘油三酯含量分彆為(0.65±0.02)mmol/L和(0.56±0.01)mmol/L,t=4.312,P<0.01,差異有統計學意義.HE染色顯示實驗組肝細胞胞質含脂肪滴減少,空泡細胞數量減少,脂肪肝得到改善.AML12肝細胞mSREBP1c錶達實驗組為0.03±0.01,對照組為0.06±0.01,t=5.624,P<0.01,差異有統計學意義.與基礎狀態相比,給以胰島素刺激後實驗組和對照組mSREBP1c錶達均增加,實驗組上升至0.06±0.02,t=8.452,P<0.01,對照組上升至0.08±0.02,t=3.591,P<0.05.高糖刺激對實驗組和對照組mSREBP1c錶達均無明顯影響.與高胰島素刺激組相比,高糖高胰島素刺激後實驗組和對照組mSREBP1c錶達均無明顯差異.結論 S6K1通過影響脂代謝的關鍵調控基因mSREBP1c錶達參與瞭脂肪的閤成,推測S6K1在脂肪肝的形成中髮揮瞭重要作用.
목적 탐토핵당체단백S6격매1(S6K1)기인침묵후대고당자격하소서간세포고순조절원건결합단백1c(SREBP1c)표체적영향.방법 장구건적S6K1shRNA중조기인선병독(S6K1Ax)주사진db/db소서미정맥,이주사함pU6계동자적선병독(pU6Ax)공재체적db/db소서작대조조,HE염색관찰간장병리개변병검측간장감유삼지함량변화.S6K1Ax전염소서간세포AML12세포,전염pU6Ax적작위대조조,분별용고당、고이도소,고당고이도소자격,역전록취합매련식반응분석간세포재각충조건자격후mSREBP1c등적표체,Western blot검측db/db소서간장급AML12세포S6K1적단백표체.결과 db/db당뇨병소서주사S6K1Ax후1주,간장S6K1단백표체피억제,대조조여실험조간장감유삼지함량분별위(0.65±0.02)mmol/L화(0.56±0.01)mmol/L,t=4.312,P<0.01,차이유통계학의의.HE염색현시실험조간세포포질함지방적감소,공포세포수량감소,지방간득도개선.AML12간세포mSREBP1c표체실험조위0.03±0.01,대조조위0.06±0.01,t=5.624,P<0.01,차이유통계학의의.여기출상태상비,급이이도소자격후실험조화대조조mSREBP1c표체균증가,실험조상승지0.06±0.02,t=8.452,P<0.01,대조조상승지0.08±0.02,t=3.591,P<0.05.고당자격대실험조화대조조mSREBP1c표체균무명현영향.여고이도소자격조상비,고당고이도소자격후실험조화대조조mSREBP1c표체균무명현차이.결론 S6K1통과영향지대사적관건조공기인mSREBP1c표체삼여료지방적합성,추측S6K1재지방간적형성중발휘료중요작용.
Objective To study the role of S6K1 in the induction of SREBPIc in mouse hepatic cell by high glucose stimulation. Methods S6K1 shRNA recombinant adenovirus (S6K1Ax) was injected into tail vein of db/db mice and then hepatic triglycerol content was analyzed. Liver specimen were stained with HE. After transfection with S6K1Ax or pU6Ax, mouse hepatic AML12 cells were treated with high glucose, insulin or glucose and insulin, the expression of mSREBP1c was detected by RT-PCR. S6K1 protein was detected by Western blot. Results Hepatic S6K1 protein in db/db mice was inhibited a week after S6K1Ax injection. Compared with the control group, hepatic triglycerol content of S6K1Ax group was decreased (0.65±0.02) mmol/L vs (0.56±0.01) mmol/L (t=4.312, P<0.01), hepatocyte fat droplet and vaculor generation were also decreased, fatty liver was improved. The mSREBP1c expression in S6K1Ax transfected cells was lower than that in the control cells (0.03±0.01 vs 0.06±0.01, t=5.624, P<0.01). Compared with the basal state, SREBP1c expression of both groups was increased on the insulin stimulation, S6K1Ax group was 0.06±0.02 (t=8.452, P<0.01) and control group was 0.08±0.02 (t=3.591,P<0.05). There is no difference between control and S6K1Ax group by glucose addition (P>0.05). Conclusion S6K1 acta on fatty synthesis by regulating mSREBP1c expression.
