米格列奈对内皮细胞一氧化氮合成及氧化应激的影响

Effects of Mitiglinide on nitric oxide production and oxidative stress in human endothelial cells

目的观察口服降糖药米格列奈对高糖环境下内皮细胞一氧化氮(NO)合成及氧化应激反应的影响。方法成功培养及传代人主动脉内皮细胞(HAECs),分为对照组(葡萄糖浓度为5.5 mmol/L)、高糖组(葡萄糖浓度为30 mmol/L)、米格列奈组(30 mmol/L葡萄糖+10μmol/L米格列奈)。ELISA法测NO、上清液中内皮型一氧化氮合酶(eNOS)及诱导型一氧化氮合酶(iNOS)的含量;比色法分别检测6、12、24、48、72 h上清中超氧化物岐化酶(SOD)的活力及丙二醛(MDA)的含量。结果①米格列奈组24 h NO含量为(232.71±8.78)μmol/L,较高糖组低(P<0.05),随后逐渐升高,48 h和72 h含量[(263.48±6.32)、(270.91±6.03)μmol/L]较高糖组明显增高,差异有统计学意义(P<0.05)。②干预24、48、72 h测米格列奈组eNOS的含量[(51.96±2.65)、(59.28±3.63)、(60.41±4.48)μmol/L]均较高糖组增高,差异有统计学意义(P<0.05);同步测iNOS含量[(53.28±3.45)、(62.09±3.71)、(59.90±3.60)μmol/L]与高糖组相比明显降低,差异有统计学意义(P<0.05)。③干预6、12、24、48、72 h测米格列奈组SOD活力[(3.99±0.46)、(5.70±0.14)、(4.98±0.37)、(3.18±0.25)、(3.35±0.51)U/mL]较高糖组高,差异有统计学意义(P<0.05);测MDA含量[(0.500±0.014)、(0.633±0.018)、(0.623±0.051)、(0.510±0.030)、(0.513±0.015)nmol/mL]较高糖组明显降低,差异有统计学意义(P<0.05)。结论米格列奈能减少高糖环境中动脉内皮细胞iNOS的生成,增强eNOS的活性和表达,促进内皮细胞NO的产生,同时能减轻高糖引起的氧化应激损伤。

Objective To study the effects of oral hypoglycemic drugs Mitiglinide on nitric oxide （NO） production and oxidative stress. Methods Human aortic endothelial cells （HAECs） were cultured and incubated successfully, they were divided into control group （5.5 mmol/L glucose）, high glucose group （30 mmol/L glucose） and Mitiglinide group （30 mmol/L glucose ＋ 10 μmol/L Mitiglinide）. NO, endothelial nitric oxide synthase （eNOS） and inducible nitric oxide synthase （iNOS） contents were measured by ELISA. SOD and MDA were examined by colorimetric method after 6, 12, 24, 48 and 72 hours respectively. Results （1） The NO content of Mitiglinide group at 24 h [（232.71±8.78） μmol/L] were lower than high glucose group, and increased after 48 and 72 hours, the NO contents [（263.48±6.32）, （270.91±6.03） μmol/L] were significantly higher than high glucose group, the differences were statistically significant （P 〈 0.05）. （2） The eNOS contents of Mitiglinide group after 24, 48 and 72 hours intervened [（51.96±2.65）, （59.28±3.63）, （60.41± 4.48）μmol/L] were higher than high glucose group, the differences were statistically significant （P 〈 0.05）. The iNOS contents of Mitiglinide group were detected at the same time, they were （53.28±3.45）, （62.09±3.71） and （59.90＋3.60） μmol/L respectively, which were significantly lower than high glucose group, the differences were statistically signifi- cant （P 〈 0.05）. （3）SOD activities of Mitiginide group were detected after 6, 12, 24, 48 and 72 hours intervened [（3.99±0.46）, （5.70±0.14）, （4.98±0.37）, （3.18±0.25） and （3.35±0.51） U/mL respectively], which were higher than high glucose group, the differences were statistically significant （P 〈 0.05）, and the MDA contents [（0.500±0.014）, （0.633± 0.018）, （0.623±0.051）, （0.510±0.030） and （0.513±0.015） nmol/mL respectively] were lower than high glucose group, the differences were statistically significant （P 〈 0.05）. Conclusion Mitiglinide is capable of decreasing the eneration of iNOS in human aortic endothelial cells with high glucose, increasing eNOS activity and its expression and promoting NO production, reducin, the iniurv from oxidative stress caused by high ducose.