Ang(1-7)减轻棕榈酸诱导的Min6细胞损伤
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摘要
目的:观察血管紧张素(1-7)[Ang(1-7)]对棕榈酸(PA)诱导小鼠胰岛β细胞株Min6功能受损的影响,并初步探讨Ang(1-7)对Min6细胞的保护作用与自噬的关系。方法:将对数生长期的Min6细胞随机分为对照(control)组、PA组、PA+Ang(1-7)组、PA+Ang(1-7)+A779组、PA+Ang(1-7)+rapamycin组、Ang(1-7)组和A779组。干预结束后用葡萄糖刺激胰岛素释放实验检测Min6细胞的分泌功能,流式细胞术检测凋亡率,活性氧簇(ROS)试剂盒检测细胞内ROS的水平,Western blot检测自噬相关蛋白LC3-I和LC3-II的表达。结果:与control组相比,PA组Min6细胞分泌功能明显受损,葡萄糖刺激后胰岛素分泌显著降低(P <0. 05),细胞凋亡率增加(P <0. 05),细胞内ROS水平和LC3-II/LC3-I比值明显增高(P <0. 05);与PA组相比,PA+Ang(1-7)组细胞葡萄糖刺激后胰岛素分泌增加(P <0. 05),细胞凋亡减少(P <0. 05),细胞内ROS水平和LC3-II/LC3-I比值均明显下降(P <0. 05)。Ang(1-7)的这一保护作用可被A779和rapamycin部分阻断。结论:Ang(1-7)减轻PA诱导的Min6细胞损伤,其细胞保护机制可能与抑制细胞自噬活性有关。
AIM: To investigate the effect of angiotensin(1-7) [Ang(1-7)]on palmitic acid(PA)-induced injury of Min6 cells and the potential protective mechanisms of autophagy. METHODS: Cultured Min6 cells were divided into 7 groups: control group,PA group,PA + Ang(1-7) group,PA + Ang(1-7) + A779 group,PA + Ang(1-7) + rapamycin group,Ang(1-7) group and A779 group. The function of Min6 cells was detected by glucose-stimulated insulin release test. Intracellular reactive oxygen species( ROS) production was measured by ROS assay kit. Apoptosis was analyzed by flow cytometry with Annexin V/PI staining. Autophagy-related proteins were determined by Western blot. RESULTS:Compared with control group,the secretion of insulin from Min6 cells in PA group was significantly decreased( P < 0. 05),and the apoptotic rate was increased( P < 0. 05). The ratio of LC3-II/LC3-I was significantly increased( P < 0. 05). Compared with PA group,the insulin secretion in PA + Ang(1-7) group was increased( P < 0. 05). The intracellular ROS level and the A779 and LC3-II/LC3-I were significantly decreased( P < 0. 05). This protective effect of Ang(1-7) was partially blocked by A779 and rapamycin. CONCLUSION: Ang(1-7) attenuates PA-induced Min6 cell injury,and its protective mechanism may be related to inhibiting the activity of autophagy.
AIM: To investigate the effect of angiotensin(1-7) [Ang(1-7)]on palmitic acid(PA)-induced injury of Min6 cells and the potential protective mechanisms of autophagy. METHODS: Cultured Min6 cells were divided into 7 groups: control group,PA group,PA + Ang(1-7) group,PA + Ang(1-7) + A779 group,PA + Ang(1-7) + rapamycin group,Ang(1-7) group and A779 group. The function of Min6 cells was detected by glucose-stimulated insulin release test. Intracellular reactive oxygen species( ROS) production was measured by ROS assay kit. Apoptosis was analyzed by flow cytometry with Annexin V/PI staining. Autophagy-related proteins were determined by Western blot. RESULTS:Compared with control group,the secretion of insulin from Min6 cells in PA group was significantly decreased( P < 0. 05),and the apoptotic rate was increased( P < 0. 05). The ratio of LC3-II/LC3-I was significantly increased( P < 0. 05). Compared with PA group,the insulin secretion in PA + Ang(1-7) group was increased( P < 0. 05). The intracellular ROS level and the A779 and LC3-II/LC3-I were significantly decreased( P < 0. 05). This protective effect of Ang(1-7) was partially blocked by A779 and rapamycin. CONCLUSION: Ang(1-7) attenuates PA-induced Min6 cell injury,and its protective mechanism may be related to inhibiting the activity of autophagy.
引文
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[5]Zhang F,Liu C,Wang L,et al.Antioxidant effect of angiotensin(1-7)in the protection of pancreaticβcell function[J].Mol Med Rep,2016,14(3):1963-1969.
[6]Sahr A,Wolke C,Maczewsky J,et al.The angiotensin-(1-7)/Mas axis improves pancreaticβ-cell function in vitro and in vivo[J].Endocrinology,2016,157(12):4677-4690.
[7]Watada H,Fujitani Y.Minireview:autophagy in pancreaticβ-cells and its implication in diabetes[J].Mol Endocrinol,2015,29(3):338-348.
[8]Masini M,Bugliani M,Lupi R,et al.Autophagy in human type 2 diabetes pancreaticβcells[J].Diabetologia,2009,52(6):1083-1086.
[9]Li L,Tan J,Miao Y,et al.ROS and autophagy:interactions and molecular regulatory mechanisms[J].Cell Mol Neurobiol,2015,35(5):615-621.
[10]Zhang X,Yu L,Xu H.Lysosome calcium in ROS regulation of autophagy[J].Autophagy,2016,12(10):1954-1955.