四川医学
四川醫學
사천의학
SICHUAN MEDICAL JOURNAL
2015年
1期
25-27,28
,共4页
GLP-1%胰岛%氧化应激%细胞凋亡%Akt磷酸化
GLP-1%胰島%氧化應激%細胞凋亡%Akt燐痠化
GLP-1%이도%양화응격%세포조망%Akt린산화
glucagon like peptide-1%islet,oxidative stress%apoptosis%Akt phosphorylation
目的：观察GLP-1对H 2 O 2所致的胰岛β细胞氧化损伤的保护作用,探讨其抵抗氧化应激诱导的细胞凋亡作用机制。方法 Ficoll法分离纯化SD大鼠胰岛β细胞,实验分为正常对照组、H 2 O 2诱导胰岛损伤组、GLP-1预处理+H 2 O 2损伤组。采用FDA/PI染色法检测细胞存活,Annexin-V/PI流式细胞术检测细胞凋亡,荧光定量PCR检测氧化应激和细胞凋亡相关基因iNOX、SOD2、Caspase-3、Bcl-2表达,Western blot检测胰岛细胞Akt、P-Akt、Caspase-3蛋白表达。观察GLP-1预处理能否减少ROS诱导的胰岛β细胞凋亡,以及在此过程中PI3K/Akt信号通路的改变。结果经 H 2 O 2损伤后胰岛β细胞的增殖活性、存活率和胰岛素分泌能力显著降低,细胞凋亡率显著升高；iNOS、Caspase-3基因表达显著升高,SOD、Bcl-2基因表达显著降低；Akt蛋白磷酸化水平明显降低, Caspase-3活性明显升高。 GLP-1预处理可显著提高β细胞的增殖活性、存活率和胰岛素分泌能力,减少细胞凋亡；显著降低iNOS、Caspase-3基因表达,上调SOD基因表达；增强Akt磷酸化,减少活化Caspase-3水平。结论 GLP-1对H 2 O 2所致的大鼠胰岛氧化应激损伤具有一定的保护作用,其作用机制与增强Akt磷酸化及抑制凋亡信号分子活化相关。
目的：觀察GLP-1對H 2 O 2所緻的胰島β細胞氧化損傷的保護作用,探討其牴抗氧化應激誘導的細胞凋亡作用機製。方法 Ficoll法分離純化SD大鼠胰島β細胞,實驗分為正常對照組、H 2 O 2誘導胰島損傷組、GLP-1預處理+H 2 O 2損傷組。採用FDA/PI染色法檢測細胞存活,Annexin-V/PI流式細胞術檢測細胞凋亡,熒光定量PCR檢測氧化應激和細胞凋亡相關基因iNOX、SOD2、Caspase-3、Bcl-2錶達,Western blot檢測胰島細胞Akt、P-Akt、Caspase-3蛋白錶達。觀察GLP-1預處理能否減少ROS誘導的胰島β細胞凋亡,以及在此過程中PI3K/Akt信號通路的改變。結果經 H 2 O 2損傷後胰島β細胞的增殖活性、存活率和胰島素分泌能力顯著降低,細胞凋亡率顯著升高；iNOS、Caspase-3基因錶達顯著升高,SOD、Bcl-2基因錶達顯著降低；Akt蛋白燐痠化水平明顯降低, Caspase-3活性明顯升高。 GLP-1預處理可顯著提高β細胞的增殖活性、存活率和胰島素分泌能力,減少細胞凋亡；顯著降低iNOS、Caspase-3基因錶達,上調SOD基因錶達；增彊Akt燐痠化,減少活化Caspase-3水平。結論 GLP-1對H 2 O 2所緻的大鼠胰島氧化應激損傷具有一定的保護作用,其作用機製與增彊Akt燐痠化及抑製凋亡信號分子活化相關。
목적：관찰GLP-1대H 2 O 2소치적이도β세포양화손상적보호작용,탐토기저항양화응격유도적세포조망작용궤제。방법 Ficoll법분리순화SD대서이도β세포,실험분위정상대조조、H 2 O 2유도이도손상조、GLP-1예처리+H 2 O 2손상조。채용FDA/PI염색법검측세포존활,Annexin-V/PI류식세포술검측세포조망,형광정량PCR검측양화응격화세포조망상관기인iNOX、SOD2、Caspase-3、Bcl-2표체,Western blot검측이도세포Akt、P-Akt、Caspase-3단백표체。관찰GLP-1예처리능부감소ROS유도적이도β세포조망,이급재차과정중PI3K/Akt신호통로적개변。결과경 H 2 O 2손상후이도β세포적증식활성、존활솔화이도소분비능력현저강저,세포조망솔현저승고；iNOS、Caspase-3기인표체현저승고,SOD、Bcl-2기인표체현저강저；Akt단백린산화수평명현강저, Caspase-3활성명현승고。 GLP-1예처리가현저제고β세포적증식활성、존활솔화이도소분비능력,감소세포조망；현저강저iNOS、Caspase-3기인표체,상조SOD기인표체；증강Akt린산화,감소활화Caspase-3수평。결론 GLP-1대H 2 O 2소치적대서이도양화응격손상구유일정적보호작용,기작용궤제여증강Akt린산화급억제조망신호분자활화상관。
Objective To investigate the protective effect of GLP-1 on H 2 O 2-induced oxidative stress damage in rat islets and its potential mechanism. Methods Pancreatic islets were isolated and purified from SD rat by Ficoll method. Three of experi-mental groups were involved:normal control group, H 2 O 2 induced islet injury group, and islets pretreated with GLP-1 plus H 2 O 2 injury group. The survival rate of islets was measured by FDA/PI fluorescent staining. The apoptosis rate of islets was determined by flow cytometry with Annexin-V/PI staining. The mRNA levels of oxidative stress and apoptosis-related genes including iNOS, SOD2, Caspase-3 and Bcl-2 were detected by real-time PCR. The protein expression of Akt, P-Akt, and caspase-3 in islets was analyzed by western blot. The protective effect of GLP-1 against oxidative stress and β-cell apoptosis was investigated, and the change of PI3K/Akt signaling pathway was also observed. Results After addition of H 2 O 2 , the proliferation, survival rate and glucose-stimulated insulin secretion of islets were significantly decreased with, while the apoptosis rate of islet was significantly in-creased. The mRNA levels of iNOS and Caspase-3 were significantly increased, while mRNA levels of SOD and Bcl-2 were signifi-cantly decreased. The phosphorylation of Akt was markedly decreased, while the activity of caspase-3 was markedly elevated. In contrast, pretreatment of GLP-1 significantly improved islets proliferation, survival and insulin secretion, and significantly reduced islets apoptosis. The mRNA levels of iNOS and caspase-3 were significantly decreased, while SOD level was significantly in-creased. The phosphorylation of Akt was markedly enhanced, while the protein level of cleaved caspase-3 was markedly de-creased. Conclusion GLP-1 was able to inhibit H 2 O 2-induced oxidative stress injury in rat islets, and its effect mechanism re-lated to the enhancement of Akt phosphorylation and the deactivation of apoptosis signaling molecules.
