胰岛素对高糖培养海马神经元凋亡及IRS/PI_3K/AKT信号通路、Bcl-2与Bax蛋白表达的影响
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摘要
目的探讨胰岛素对高糖培养海马神经元的保护作用及其作用机制。方法选择新生24h SD大鼠,取海马神经元进行原代培养,分为正常对照组(NC组),高糖模型组(HG组),实验组(RI组),采用Tunel检测海马神经元的凋亡,Western blot法检测各组P-IRS、IRS1、P-AKT、AKT、Bcl-2、Bax蛋白的表达。结果与NC组相比,HG组海马神经元凋亡率显著增加(P<0.01);与HG组相比,RI组凋亡率显著降低(P<0.01)。与NC组相比,HG组P-IRS、IRS1蛋白表达显著降低(P<0.05);RI组P-IRS表达及P-IRS/IRS1显著升高(P<0.01)。与HG组相比,RI组P-IRS表达及P-IRS/IRS1显著升高(P<0.01)。与NC组相比,HG组AKT表达显著降低(P<0.01);RI组P-AKT、AKT表达及P-AKT/AKT显著升高(P<0.01)。与HG组相比,RI组P-AKT、AKT表达及P-AKT/AKT显著升高(P<0.01)。与NC组相比,HG组Bcl-2、Bcl-2/Bax蛋白表达显著降低(P<0.01);RI组Bcl-2、Bcl-2/Bax蛋白表达显著升高(P<0.01),Bax蛋白表达显著降低(P<0.01)。与HG组相比,RI组Bcl-2、Bcl-2/Bax蛋白表达显著升高(P<0.01),bax蛋白表达显著降低(P<0.01)。结论胰岛素能够抑制高糖培养海马神经元的凋亡,其作用可能是通过活化IRS1/PI3K/AKT信号通路,同时上调Bcl-2/Bax实现的。
Objective To explore the protective effect and mechanism of insulin on hippocampal neurons cultured in high glucose.Methods Hippocampus were obtained from newborn 24 h SD rats and then Primary cultured.According to the purpose of the study,hippocampal neurons were divided into 3 groups: normal control group(NC group),high glucose group(HG group),experimental group(RI group).We detected the apoptosis of neurons with TUNEL.And the expression of P-IRS,IRS1,P-AKT,AKT,Bcl-2 and Bax was detected by using western blot.Results Compared with the NC group,the apoptosis rate of hippocampal neurons in HG group was increased significantly(P < 0.01).Compared with the HG group,the apoptosis rate of hippocampal neurons in RI group was decreased significantly(P < 0.01).Compared with the NC group,the expression of P-IRS and IRS1 of HG group was decreased significantly(P <0.05),the expression of P-IRS and P-IRS/IRS1 of RI group was increased significantly(P < 0.01).Compared with the HG group,the expression of P-IRS and P-IRS/IRS1 of RI group was increased significantly(P < 0.01).Compared with the NC group,the expression of AKT of HG group was decreased significantly(P < 0.01),the expression of P-AKT,AKT and P-AKT/AKT of RI group was increased significantly(P < 0.01).Compared with the HG group,the expression of P-AKT,AKT and P-AKT/AKT of RI group was increased significantly(P < 0.01).Compared with the NC group,the expression of Bcl-2 and Bcl-2/Bax of HG group was decreased significantly(P < 0.01),the expression of Bcl-2 and Bcl-2/Bax of RI group was increased significantly(P < 0.01),and the expression of bax was decreased significantly(P < 0.01).Compared with the HG group,the expression of Bcl-2 and Bcl-2/Bax of RI group was increased significantly(P < 0.01),and the expression of bax was decreased significantly(P < 0.01).Conclusion Insulin could reduce the apoptosis of neurons which cultured in high glucose by activating the IRS1/PI3K/AKT signal pathway and up-regulating Bcl-2/Bax.
Objective To explore the protective effect and mechanism of insulin on hippocampal neurons cultured in high glucose.Methods Hippocampus were obtained from newborn 24 h SD rats and then Primary cultured.According to the purpose of the study,hippocampal neurons were divided into 3 groups: normal control group(NC group),high glucose group(HG group),experimental group(RI group).We detected the apoptosis of neurons with TUNEL.And the expression of P-IRS,IRS1,P-AKT,AKT,Bcl-2 and Bax was detected by using western blot.Results Compared with the NC group,the apoptosis rate of hippocampal neurons in HG group was increased significantly(P < 0.01).Compared with the HG group,the apoptosis rate of hippocampal neurons in RI group was decreased significantly(P < 0.01).Compared with the NC group,the expression of P-IRS and IRS1 of HG group was decreased significantly(P <0.05),the expression of P-IRS and P-IRS/IRS1 of RI group was increased significantly(P < 0.01).Compared with the HG group,the expression of P-IRS and P-IRS/IRS1 of RI group was increased significantly(P < 0.01).Compared with the NC group,the expression of AKT of HG group was decreased significantly(P < 0.01),the expression of P-AKT,AKT and P-AKT/AKT of RI group was increased significantly(P < 0.01).Compared with the HG group,the expression of P-AKT,AKT and P-AKT/AKT of RI group was increased significantly(P < 0.01).Compared with the NC group,the expression of Bcl-2 and Bcl-2/Bax of HG group was decreased significantly(P < 0.01),the expression of Bcl-2 and Bcl-2/Bax of RI group was increased significantly(P < 0.01),and the expression of bax was decreased significantly(P < 0.01).Compared with the HG group,the expression of Bcl-2 and Bcl-2/Bax of RI group was increased significantly(P < 0.01),and the expression of bax was decreased significantly(P < 0.01).Conclusion Insulin could reduce the apoptosis of neurons which cultured in high glucose by activating the IRS1/PI3K/AKT signal pathway and up-regulating Bcl-2/Bax.
引文
1 Arvanitakis Z,Wilson RS,Bienias JL,et al.Diabetes mellitus and risk of Alzheimer disease and decline in cognitive function[J].Arch Neurol,2004,61(5):661-666
2 Zhang YW,Zhang JQ,Liu C,et al.Memory dysfunction in type 2 diabetes mellitus correlates with reduced hippocampal CA1 and subiculum volumes[J].Chinese Med J,2015,128(4):465-471
3 Banks WA,Owen JB,Erickson MA.Insulin in the brain:there and back again[J].Pharmacol Thera peut,2012,136(1):82-93
4傅静奕,李焱,严励,等.NF-κB在甘精胰岛素拮抗胰岛β细胞脂性凋亡中的作用[J].中国药理学通报,2007,23(8):1048-1052
5葛晨亮,刘厂辉,唐振旺,等.胰岛素对大鼠平滑肌细胞表达MMP-9、TIMP-1及内皮细胞表达s ES的影响[J].中国动脉硬化杂志,2016,24(2):145-149
6陈永松,张娟,朱旭新,等.胰岛素对高葡萄糖浓度下内皮细胞的凋亡及bax和bcl-2表达的影响[J].中国临床实用医学,2007,1(8):1
7叶瑾.胰岛素通过Akt/Fox O/SVV信号通路抑制缺血/再灌注损伤诱导的心肌微血管内皮细胞凋亡的研究[D].西安:第四军医大学,2014
8王华梅,周青珍,吴家幂.胰岛素对大鼠脑缺血再灌注后Bcl-2表达及细胞凋亡的影响[J].神经疾病与精神卫生,2011,11(4):333-336
9李志飞,张建明,刘群,等.胰岛素对烟雾吸入性损伤大鼠肺组织细胞凋亡及Bcl-2和Bax蛋白表达的影响[J].中华损伤与修复杂志:电子版,2011,6(2):31-34
10徐敏.高糖刺激对雪旺细胞增殖与凋亡机制的探讨[D].济南:山东大学,2014
11郭丽萍,王坚,蒋雨平.磷脂酰肌醇3-激酶/蛋白激酶B磷酸化是胰岛素受体后信号转导通路控制PC12细胞凋亡的机制[J].中国临床神经科学,2007,15(6):596-599
12姜寿峰,边连防,陈晓红,等.胰岛素对谷氨酸诱导PC12细胞损伤的保护作用[J].中国老年学杂志,2005,25(1):62-64
13 Sun X,Huang L,Zhang M,et al.Insulin like growth factor-1 prevents 1-mentyl-4-phenyl phyridinium-induced apoptosis in PC12 cells through activation of glycogen synthase kinase-3beta[J].Toxicology,2010,271(1-2):5-12
14 Song G,Ouyang G,Bao S.The activation of Akt/PKB signaling pathway and cell survival[J].J Cell Mol Med,2005,9(1):59-71
15 Kern W,Peters A,Fruehwald Schultes B,et al.Improving influence of insulin on cognitive functions in humans[J].Neuroendocrinology,2001.74(4):270-280
2 Zhang YW,Zhang JQ,Liu C,et al.Memory dysfunction in type 2 diabetes mellitus correlates with reduced hippocampal CA1 and subiculum volumes[J].Chinese Med J,2015,128(4):465-471
3 Banks WA,Owen JB,Erickson MA.Insulin in the brain:there and back again[J].Pharmacol Thera peut,2012,136(1):82-93
4傅静奕,李焱,严励,等.NF-κB在甘精胰岛素拮抗胰岛β细胞脂性凋亡中的作用[J].中国药理学通报,2007,23(8):1048-1052
5葛晨亮,刘厂辉,唐振旺,等.胰岛素对大鼠平滑肌细胞表达MMP-9、TIMP-1及内皮细胞表达s ES的影响[J].中国动脉硬化杂志,2016,24(2):145-149
6陈永松,张娟,朱旭新,等.胰岛素对高葡萄糖浓度下内皮细胞的凋亡及bax和bcl-2表达的影响[J].中国临床实用医学,2007,1(8):1
7叶瑾.胰岛素通过Akt/Fox O/SVV信号通路抑制缺血/再灌注损伤诱导的心肌微血管内皮细胞凋亡的研究[D].西安:第四军医大学,2014
8王华梅,周青珍,吴家幂.胰岛素对大鼠脑缺血再灌注后Bcl-2表达及细胞凋亡的影响[J].神经疾病与精神卫生,2011,11(4):333-336
9李志飞,张建明,刘群,等.胰岛素对烟雾吸入性损伤大鼠肺组织细胞凋亡及Bcl-2和Bax蛋白表达的影响[J].中华损伤与修复杂志:电子版,2011,6(2):31-34
10徐敏.高糖刺激对雪旺细胞增殖与凋亡机制的探讨[D].济南:山东大学,2014
11郭丽萍,王坚,蒋雨平.磷脂酰肌醇3-激酶/蛋白激酶B磷酸化是胰岛素受体后信号转导通路控制PC12细胞凋亡的机制[J].中国临床神经科学,2007,15(6):596-599
12姜寿峰,边连防,陈晓红,等.胰岛素对谷氨酸诱导PC12细胞损伤的保护作用[J].中国老年学杂志,2005,25(1):62-64
13 Sun X,Huang L,Zhang M,et al.Insulin like growth factor-1 prevents 1-mentyl-4-phenyl phyridinium-induced apoptosis in PC12 cells through activation of glycogen synthase kinase-3beta[J].Toxicology,2010,271(1-2):5-12
14 Song G,Ouyang G,Bao S.The activation of Akt/PKB signaling pathway and cell survival[J].J Cell Mol Med,2005,9(1):59-71
15 Kern W,Peters A,Fruehwald Schultes B,et al.Improving influence of insulin on cognitive functions in humans[J].Neuroendocrinology,2001.74(4):270-280