橙皮素对高糖所致血管内皮功能障碍和血管内皮细胞损伤的保护作用

Protective effect of hesperetin on hyperglycemia-induced vascular endothelial dysfunction and vascular endothelial cell injury

目的研究橙皮素(HSP)对高糖(44 mmol·L^(-1))诱导的血管舒张功能障碍和血管内皮细胞损伤的影响及其作用机制。方法将大鼠冠状动脉(RCA)血管环和人脐静脉内皮细胞(HUVECs)分为6组:正常对照组(生理盐溶液)、高渗对照组(甘露醇44 mmol·L^(-1))、模型组(葡萄糖44 mmol·L^(-1))和低、中、高3个剂量实验组(在模型组基础上分别加橙皮素10,30,100μmol·L^(-1))。血管张力法测定HSP对RCA舒张功能的影响,微孔板法检测HUVECs的丙二醛(MDA)、一氧化氮(NO)、乳酸脱氢酶(LDH)释放量、超氧化物歧化酶(SOD)及一氧化氮合酶(NOS)活性。结果以血栓素A2受体激动药U46619 (1μmol·L^(-1))预收缩RCA时,正常对照组、高渗对照组、模型组和低、中、高3个剂量实验组对乙酰胆碱的最大舒张率分别为(83.45±6.31)%,(85.60±4.50)%,(52.30±5.87)%,(68.92±8.88)%,(71.60±5.30)%和(81.60±6.39)%;这6组的LDH释放量分别为(332±88),(345±92),(856±113),(568±102),(485±96)和(469±73) U·L^(-1);SOD活性分别为(56.6±7.6),(53.3±5.9),(28.7±6.3),(33.6±4.8),(45.3±7.7)和(52.6±6.5) kU·L^(-1);NOS活性分别为(11.3±0.48),(12.6±0.22),(5.8±0.31),(6.6±0.52),(9.8±0.29)和(12.6±0.51) kU·g^(-1)。模型组与正常对照组相比,或中、高2个剂量实验组与模型组相比,上述指标的差异均有统计学意义(均P <0.05)。结论 HSP减轻高糖所致的RCA内皮依赖性舒张功能障碍,其机制可能与HSP抗氧化作用及增加NOS活性有关。

Objective To study the effects and mechanism of hesperetin(HSP) on vasodilator impairment and endothelial injury induced by high glucose.Methods Rat coronary artery(RCA) rings and human umbilical vein endothelial cells(HUVECs) were divided into six groups:normal control group(physiological saline solution);hyperosmotic control group(mannitol 44 mmol·L^(-1));model group(glucose 44 mmol·L^(-1));experimental-L,experimental-M and experimental-H groups(10,30,100 μmol·L^(-1) HSP basis on model group,respectively).A wire myograph was used to record the effect of HSP on the diastolic function of RCA.The release of malondialdehyde(MDA),nitric oxide(NO) and lactate dehydrogenase(LDH),superoxide dismutase(SOD) and nitric oxide synthase(NOS) activity of HUVECs were measured by microwell plate method.Results When thromboxane A2 receptor agonist U46619(1 μmol·L^(-1))precontracted RCA rings,the maximum relaxation rates to acetylcholine(Ach) in normal control group,hyperosmotic control group,model group;experimental-L,experimental-M and experimental-H groups were(83.45±6.31) %,(85.60±4.50) %,(52.30±5.87) %;(68.92±8.88) %,(71.60±5.30) % and(81.60±6.39) %,respectively;the release of LDH in the six groups were(332±88),(345±92),(856±113);(568±102),(485±96),(469±73) U·L^(-1),respectively;the activity of SOD in the six groups were(56.6±7.6),(53.3±5.9),(28.7±6.3),(33.6±4.8),(45.3±7.7),(52.6±6.5) kU·L^(-1),respectively;the activity of NOS in the six groups were(11.3±0.48),(12.6±0.22),(5.8±0.31),(6.6±0.52),(9.8±0.29),(12.6±0.51) kU.g^(-1),respectively.Compared between model group and normal control group,or experimental-M and experimental-H groups and model group,the differences of the factors were statistically significant(All P <0.05).Conclusion HSP alleviates high glucose-induced impairment in endotheliumdependent vasodilation of RCA,which may be related to the antioxidant effect of HSP and the increase of NOS activity.