中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
20期
1413-1416
,共4页
陈说%曹筱佩%肖海鹏%莫小庆%李延兵
陳說%曹篠珮%肖海鵬%莫小慶%李延兵
진설%조소패%초해붕%막소경%리연병
胰高血糖素%胰岛%细胞凋亡%脂肪酸类,非酯化
胰高血糖素%胰島%細胞凋亡%脂肪痠類,非酯化
이고혈당소%이도%세포조망%지방산류,비지화
Glucagon%Islets of langerhans%Cell apoptosis%Fatty aoids,nonesterified
目的 观察游离脂肪酸(FFA)对胰岛β-TC3细胞胰腺衍生因子(PANDER)表达的影响,并探讨胰高血糖素样多肽1(GLP-1)对其的拮抗作用及与蛋白激酶B(Akt)信号通路的关系.方法 培养β-TC3细胞,以不同浓度棕榈酸(PA)处理12 h,0.5 mmol/L的PA处理不同时间后实时荧光定量PCR法检测PANDER mRNA表达;Western印迹检测对照组、PA组、PA+GLP-1组、GLP-1组处理12 h后PANDER、P-Akt及半胱氨酸天冬氨酸蛋白酶-3酶原蛋白表达;在上述各组加入Akt特异阻断剂(Akti-1/2)后,再检测PANDER蛋白表达,并以Hoechst33258核染色法检测细胞凋亡情况.结果 (1)PA浓度依赖性及时间依赖性增加PANDER mRNA表达(P<0.05);(2)与对照组(100%)比较,PA组诱导PANDER蛋白表达高[(148±18)%,P<0.05),PA+GLP-1组PA诱导PANDER表达为[(70±17)%,P<0.01];(3)Akti-1/2减弱GLP-1抑制PA诱导的PANDER表达及细胞凋亡的效应:PA+GLP-1+Akti-1/2组较PA+GLP-1组PANDER蛋白显著高[(249+49)%比(110±54)%,P<0.01],细胞凋亡率显著高[(37.8±-1.5)%比(20.1±3.5)%,P<0.01].结论 PA可诱导PANDER的表达及胰岛B细胞凋亡,GLP-1通过激活Akt通路拮抗PA诱导的PANDER表达及胰岛B细胞凋亡.
目的 觀察遊離脂肪痠(FFA)對胰島β-TC3細胞胰腺衍生因子(PANDER)錶達的影響,併探討胰高血糖素樣多肽1(GLP-1)對其的拮抗作用及與蛋白激酶B(Akt)信號通路的關繫.方法 培養β-TC3細胞,以不同濃度棕櫚痠(PA)處理12 h,0.5 mmol/L的PA處理不同時間後實時熒光定量PCR法檢測PANDER mRNA錶達;Western印跡檢測對照組、PA組、PA+GLP-1組、GLP-1組處理12 h後PANDER、P-Akt及半胱氨痠天鼕氨痠蛋白酶-3酶原蛋白錶達;在上述各組加入Akt特異阻斷劑(Akti-1/2)後,再檢測PANDER蛋白錶達,併以Hoechst33258覈染色法檢測細胞凋亡情況.結果 (1)PA濃度依賴性及時間依賴性增加PANDER mRNA錶達(P<0.05);(2)與對照組(100%)比較,PA組誘導PANDER蛋白錶達高[(148±18)%,P<0.05),PA+GLP-1組PA誘導PANDER錶達為[(70±17)%,P<0.01];(3)Akti-1/2減弱GLP-1抑製PA誘導的PANDER錶達及細胞凋亡的效應:PA+GLP-1+Akti-1/2組較PA+GLP-1組PANDER蛋白顯著高[(249+49)%比(110±54)%,P<0.01],細胞凋亡率顯著高[(37.8±-1.5)%比(20.1±3.5)%,P<0.01].結論 PA可誘導PANDER的錶達及胰島B細胞凋亡,GLP-1通過激活Akt通路拮抗PA誘導的PANDER錶達及胰島B細胞凋亡.
목적 관찰유리지방산(FFA)대이도β-TC3세포이선연생인자(PANDER)표체적영향,병탐토이고혈당소양다태1(GLP-1)대기적길항작용급여단백격매B(Akt)신호통로적관계.방법 배양β-TC3세포,이불동농도종려산(PA)처리12 h,0.5 mmol/L적PA처리불동시간후실시형광정량PCR법검측PANDER mRNA표체;Western인적검측대조조、PA조、PA+GLP-1조、GLP-1조처리12 h후PANDER、P-Akt급반광안산천동안산단백매-3매원단백표체;재상술각조가입Akt특이조단제(Akti-1/2)후,재검측PANDER단백표체,병이Hoechst33258핵염색법검측세포조망정황.결과 (1)PA농도의뢰성급시간의뢰성증가PANDER mRNA표체(P<0.05);(2)여대조조(100%)비교,PA조유도PANDER단백표체고[(148±18)%,P<0.05),PA+GLP-1조PA유도PANDER표체위[(70±17)%,P<0.01];(3)Akti-1/2감약GLP-1억제PA유도적PANDER표체급세포조망적효응:PA+GLP-1+Akti-1/2조교PA+GLP-1조PANDER단백현저고[(249+49)%비(110±54)%,P<0.01],세포조망솔현저고[(37.8±-1.5)%비(20.1±3.5)%,P<0.01].결론 PA가유도PANDER적표체급이도B세포조망,GLP-1통과격활Akt통로길항PA유도적PANDER표체급이도B세포조망.
Objective To investigate the effects of free fatty acid(FFA)on the expression of pancreatic derived factor(PANDER)in β-cells and to explore the possible relationship between PANDER and Akt signaling pathway at the anti-apoptotic effects of glucagon-like peptide-l(GLP-1).Methods β-TC3 cells were cultured in vitro witH palmitic acid(PA)of different concentrations and different time courses.The expression of PANDER mRNA was analyzed by realtime quantitative PCR β-TC3 cells were cultured with vehicle.0.5 mmol/L PA.0.5 mmol/L PA+10 nmol/L GLP-1 and 10nmol/L GLP-1respectively with or without Akti-1/2, a selective inhibitor of Akt, for 12 hours.The protein levels of PANDER, P-Akt and pro-caspase3 were detected by Western blot And cell apoptosis was analyzed by Hoechst33258 staining.Results (1)PA could dose and time dependently increased the expression of PANDER mRNA in β cells(vs control, P<0.05);(2)PA increased the PANDER protein expression [(148±18)%vs control 100%.P<0.05].However,these effects were attenuated by GLP-1 [(70±17)%vs PA group,P<0.01];(3)Akt inhibitor-1/2 alleviated the effects of GLP-1 on PA inducing the expression of PANDER.The expression of PANDER increased significantly in PA+GLP-1+Akti-1/2 group [(249±49)%vs PA+GLP-1 group(110±54)%,P<0.01],and cell apoptosis increased significantly as well[(37.8±1.5)%vs PA+GLP-1 group(20.1±3.5)%,P<0.01].Conclusion PA induces the expression of PANDER and the apoptosis of β cell while GLP-1 counteracts the above effects through an activation of Akt signaling pathway.
