CAJ | 학술논문

目的探讨葡萄糖浓度波动损伤牛血管内皮细胞的机制.方法取新生牛胸主动脉进行血管内皮细胞的原代培养.实验分3组:对照组、高糖组和波动组.荧光偏振法检测3组内皮细胞的细胞膜流动性;生化方法测定细胞内山梨醇、醛糖还原酶、山梨醇脱氢酶和糖基化终产物(AGEs)的含量;逆转录聚合酶链反应法检测细胞一氧化氮合酶(eNOS)、内皮素1(ET-1)以及糖基化终产物受体mRNA的表达情况.结果偏正度和膜微黏度均为波动组和高糖组高于对照组(均P<0.01),波动组高于高糖组(均P<0.01).细胞内山梨醇含量、醛糖还原酶、AGEs含量均为波动组和高糖组高于对照组(均P<0.01),醛糖还原酶、AGEs含量均为波动组高于高糖组(均P<0.01).细胞内山梨醇脱氢酶含量波动组和高糖组均低于对照组(P<0.05),波动组低于高糖组(P<0.05).与对照组比较,高糖组和波动组细胞RAGE、eNOS和ET-1的mRNA表达均明显上调(P<0.01).结论葡萄糖浓度波动导致血管内皮细胞的细胞膜流动性降低,激活多元醇通路途径相关酶的活性,增加细胞内AGEs的积聚,上调内皮细胞eNOS、ET-1和糖基化终产物受体mRNA的表达.葡萄糖浓度波动对血管内皮细胞的损伤比持续高糖更为严重.
목적 탐토포도당농도파동손상우혈관내피세포적궤제.방법 취신생우흉주동맥진행혈관내피세포적원대배양.실험분3조:대조조、고당조화파동조.형광편진법검측3조내피세포적세포막류동성;생화방법측정세포내산리순、철당환원매、산리순탈경매화당기화종산물(AGEs)적함량;역전록취합매련반응법검측세포일양화담합매(eNOS)、내피소1(ET-1)이급당기화종산물수체mRNA적표체정황.결과 편정도화막미점도균위파동조화고당조고우대조조(균P<0.01),파동조고우고당조(균P<0.01).세포내산리순함량、철당환원매、AGEs함량균위파동조화고당조고우대조조(균P<0.01),철당환원매、AGEs함량균위파동조고우고당조(균P<0.01).세포내산리순탈경매함량파동조화고당조균저우대조조(P<0.05),파동조저우고당조(P<0.05).여대조조비교,고당조화파동조세포RAGE、eNOS화ET-1적mRNA표체균명현상조(P<0.01).결론 포도당농도파동도치혈관내피세포적세포막류동성강저,격활다원순통로도경상관매적활성,증가세포내AGEs적적취,상조내피세포eNOS、ET-1화당기화종산물수체mRNA적표체.포도당농도파동대혈관내피세포적손상비지속고당경위엄중.
Objective To explore the effects of glucose concentration fluctuation on function of cultured bovine arterial endothelial cells and underlying mechanism. Methods The thoracic aorta of newborn calf was used for primary endothelial cells culture. Cells were divided into 3 groups and cultured for 48 h : control group ( C, 5.5 mmol/L), constant high glucose group ( HG, 30 mmol/L) and glucose fluctuation ( GF, three circles of 2 h 30 mmol/L followed by 3 h 5.5 mmol/L, 30 mmol/L overnight, repeat the whole procedure on the following day) groups. The membranes fluidity of endothelial cells was detected by fluorescence polarization method. The contents of sorbierite, aldose reductase (AR), sorbitoi dehydrogenase (SDH) and advanced glycation end products (AGEs) were measured. RAGE, eNOS and ET-1 mRNA expressions were detected by semi-quantitative RT-PCR. Results The membranes fluidity of endothelial cells in HG or GF group were significantly decreased compared with the control group( all P <0.01 ) and significantly lower in GF group than those in HG group (all P < 0. O1 ). Sorbierite, AR and AGEs concentrations were significantly higher in HG and GF groups than those in control group ( all P <0. 01 ) and AR and AGEs concentrations were significantly higher in GF group than that in HG group ( all P <0.01 ). SDH of endothelial cells in HG or GF group were decreased compared with the control group and lower in GF group than in HG group ( all P < 0.05). In addition, the mRNA levels of RAGE, eNOS and ET-1 were significantly upregulated compared with the control group( all P < 0.01 ). Conclusions Glucose concentration fluctuation can result in more severe bovine arterial endothelial cells dysfunction than high glucose via activating polyols metabolic pathways, upregulating the expression of AGEs, eNOS and ET-1.Therefore, glucose concentration fluctuation might play a crucial role on macrovascular complications of diabetes.