CAJ | 학술논문

目的探讨胰岛素及磷脂酰肌醇3-激酶(PI3-K)途径对NO生成的影响.方法检测胰岛素、葡萄糖以及PI3-K活性不可逆的抑制剂(Wortmannin)对培养的人脐静脉内皮细胞(HUVECs)PI3-K表达以及NO、超氧阴离子(O2-)产生和内皮型一氧化氮合酶(eNOS)活性的影响.实验分为对照组、10 mU/L胰岛素组、100 mU/L胰岛素组、甘露醇组、5 mmol/L葡萄糖+10 mU/L胰岛素组(5 mmol/L G1组)、25 mmol/L葡萄糖+100 mU/L胰岛素组(25 mmol/L G2组)、50 nmol/L Wortmannin组(50 nmol/L W组)、50 nmol/L Wortmannin+10 mU/L胰岛素组(50 nmol/L W1组)和50 nmol/L Wortmannin+100 mU/L胰岛素组(50 nmol/L W2组).结果与对照组比较,不同浓度胰岛素组eNOS活性及NO水平显著升高(P<0.01);25 mmol/L G2组、50 nmol/L W组、50 nmol/LW1组和50 nmol/L W2组eNOS活性及NO水平均显著降低,O2-生成明显增加(P<0.01);与时照组比较,不同浓度胰岛组、50 nmol/L W组、50 nmol/L W1组和50 nmol/L W2组PI3-K蛋白表达显著升高(P<0.05,P<0.01).结论 PI3-K信号途径对于促进NO产生、维持血管内皮细胞的正常功能具有重要作用,在高糖、高胰岛素状态下该条途径受损并由此引发内皮功能障碍.
목적 탐토이도소급린지선기순3-격매(PI3-K)도경대NO생성적영향.방법 검측이도소、포도당이급PI3-K활성불가역적억제제(Wortmannin)대배양적인제정맥내피세포(HUVECs)PI3-K표체이급NO、초양음리자(O2-)산생화내피형일양화담합매(eNOS)활성적영향.실험분위대조조、10 mU/L이도소조、100 mU/L이도소조、감로순조、5 mmol/L포도당+10 mU/L이도소조(5 mmol/L G1조)、25 mmol/L포도당+100 mU/L이도소조(25 mmol/L G2조)、50 nmol/L Wortmannin조(50 nmol/L W조)、50 nmol/L Wortmannin+10 mU/L이도소조(50 nmol/L W1조)화50 nmol/L Wortmannin+100 mU/L이도소조(50 nmol/L W2조).결과 여대조조비교,불동농도이도소조eNOS활성급NO수평현저승고(P<0.01);25 mmol/L G2조、50 nmol/L W조、50 nmol/LW1조화50 nmol/L W2조eNOS활성급NO수평균현저강저,O2-생성명현증가(P<0.01);여시조조비교,불동농도이도조、50 nmol/L W조、50 nmol/L W1조화50 nmol/L W2조PI3-K단백표체현저승고(P<0.05,P<0.01).결론 PI3-K신호도경대우촉진NO산생、유지혈관내피세포적정상공능구유중요작용,재고당、고이도소상태하해조도경수손병유차인발내피공능장애.
Objective To elucidate the effect of insulin/phosphatidylinositol 3-kinase(PI3-K) sig-naling pathway on endothelial nitric oxide(NO) production. Methods The effects of insulin,glu-cose and wortmannin(inhibitor of PI3-K) on the PI3-K expression, the generation of NO and su-peroxide anion(O2-) in human umbilical vein endothelial cells(HUVECs) were investigated. The experiment included 9 groups, control group, 10 mU/L insulin, 100 mU/L insulin, mannitol, 5 mmol/L glucose+10 mU/L insulin,25 mmol/L glucose+ 100 mU/L insulin, 50 nmol/L wort-mannin, 50 nmol/L wortmannin+ 10 mU/L insulin and 50 nmol/L wortmannin+100 mU/L insu-lin. Results Different dosages of insulin(10 mU/L, 100 mU/L) significantly increased NO pro-duction and endothelial nitric oxide synthase(eNOS) activity in HUVECs as compared with con-trol group. In the presence of 25 mmol/L glucose+100 mU/L insulin,wortmannin and wortman-nin+insulin(10 mU/L, 100 mU/L), NO production and eNOS activity significantly decreased (P < 0.01 vs controls,respectively) accompanied by elevated O2- production (P < 0.01 vs con-trols, respectively). Compared with controls, insulin (10 mU/L, 100 mU/L)and wortmannin,wortmannin+insulin(10 mU/L,100 mU/L) promoted PI3-K expression. Conclusions PI3-K sig-naling pathway plays an important role in NO production and maintenance of endothelial func-tion. In insulin-resistant state, this pathway is damaged, leading to endothelial dysfunction.