血管紧张素Ⅱ对小鼠胰岛素分泌细胞株NIT-1胰岛素信号通路的影响  

Effects of angiotensin Ⅱ on insulin signaling pathway in NIT-1 cells

作　　者：徐明彤[1] 张莉[1] 薛声能[1] 姜力丹[1] 何爽[1] 柴佳妮[1] 严励[1] 

机构地区：[1]中山大学孙逸仙纪念医院内分泌科,广州510120

出　　处：《中华糖尿病杂志》2011年第2期142-146,共5页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：广东省科技计划项目（20098030801017）

摘　　要：目的 探讨血管紧张素Ⅱ对小鼠胰岛β细胞内胰岛素信号通路的影响及可能机制.方法 小鼠胰岛素分泌细胞株NIT-1以低糖培养基和无血清培养液培养后分为6组：血管紧张素Ⅱ组（A组）、胰岛素组（B组）、血管紧张素Ⅱ＋胰岛素组（C组）、血管紧张素Ⅱ＋胰岛素＋沙拉新组（D组）、血管紧张素Ⅱ＋胰岛素＋二联苯碘组（E组）、对照组（F组）.应用Western blot检测酪氨酸磷酸化胰岛素受体β亚单位、酪氨酸磷酸化胰岛素受体底物-1、丝氨酸磷酸化胰岛素受体底物-1及P47phox蛋白表达水平,采用流式细胞仪检测细胞内H2O2水平,选用逆转录-聚合酶链反应检测胰岛素mRNA水平.组间比较采用方差分析,两两比较采用LSD检验.结果 A组丝氨酸磷酸化胰岛素受体底物-1较F组显著升高（分别为1.18±0.10和0.59±0.11,LSD检验,P＜0.01）.与B组比较,C组酪氨酸磷酸化胰岛素受体β亚单位（分别为1.22±0.26和1.95±0.19）、酪氨酸磷酸化胰岛素受体底物-1（分别为0.74±0.18和1.25±0.23）明显降低（均P＜0.01）,丝氨酸磷酸化胰岛素受体底物-1显著增加（分别为1.11±0.17和0.62±0.10,P＜0.01）.D组、E组酪氨酸磷酸化胰岛素受体β亚单位、酪氨酸磷酸化胰岛素受体底物-1较C组升高,丝氨酸磷酸化胰岛素受体底物-1 下降.A组、C组细胞内H2O2显著高于F组（分别为44.2±2.2、41.0±5.0和28.6±1.7,均P＜0.01）,D组、E组低于C组（分别为30.2±1.7、30.8±2.1和41.0±5.0,均P＜0.01）.A组、C组P47 phox水平明显高于F组（分别为1.62±0.17、1.59±0.19和0.89±0.12,均P＜0.01）,D组、E组P47 phox水平显著低于C组（分别为1.09±0.16、30.8±2.1和1.59±0.19,均P＜0.01）.就胰岛素mRNA表达水平而言,B组显著高于F组（分别为0.90±0.15和0.60±0.19,P＜0.01）,C组低于B组（分别为0.62±0.19和0.90±0.15,P＜0.01）,D组、E组明显高于C组（分别为0.80±0.17、0.82±0.19和0.62±0.19,均P＜0.0Objective To evaluate the effects of angiotensin Ⅱ on insulin signaling pathway in insulin-secreting NIT-1 cells in mice. Methods NIT-1 eells were cultured in the modified DMED,and then in the serum-free DMEM before assigned to the following 6 groups： angiotensin Ⅱ group （ group A ）,insulin group （ group B） ,angiotensin Ⅱ ＋ insulin group （ group C ） ,angioteusin Ⅱ ＋insulin ＋ saralasin group （group D ）,angiotensin Ⅱ ＋ insulin ＋ Diphenyleneiodonium group （group E ） and control group （group F）. Expression of insulin receptor tyrosine phosphorylation （IR-β-Tyr） ,tyrosine phosphorylation of insulin receptor substrate-1 （IRS-1-Tyr）,serine phosphorylation of insulin receptor substrate-1 （IRS-1-Ser）,and P47 phox were evaluated by Western blot Level of H2O2 was detected by flow cytometry.Expression of insulin mRNA was evaluated by reverse transcript-polymerase chain reaction. One-way analysis of variance （ANOVA） and LSD test were used for data analysis. Results Expression of IRS-1-Ser was higher in the group A than group F. The levels of IR-β-Tyr （1.22 ±0.26 vs 1.95 ±0. 19,LSD test,P〈0. 01 ） and IRS-1-Tyr （0. 74 ±0. 18 vs 1.25 ±0. 23,P 〈0. 01 ） in group C were lower than those in group B,whereas the level of IRS-Ser was higher （ 1. 11 ± 0. 17 vs 0. 62 ± 0. 10,P 〈 0. 01 ）. The expressions of IR-β-Tyr and IRS-1-Tyr in groups D and E were significantly increased when compared with group C,and the level of IRS-1-Ser changed adversely. The levels of H2O2 and P47 phox in groups A and C were significantly increased when compared with group F,and those in groups D and E were lower in comparison with group C. The expression of insulin mRNA was significantly increased in groups D and E than group C （ group D vs group C： 0. 80 ± 0. 17 vs 0. 62 ± 0. 19,P 〈 0. 05; group Evs group C： 0. 82 ± 0. 19 vs 0. 62 ±0. 19,P 〈 0. 01 ）. Conclusions Angiotensin Ⅱ could increase the expression of IRS-1-Ser and inhibit insulin-stimulated IR-β-

关 键 词：血管紧张素Ⅱ 受体 胰岛素 氧化性应激 

分 类 号：R587.1[医药卫生—内分泌]