CAJ | 학술논문

目的探讨牛磺酸对高糖诱导的内皮细胞凋亡的影响及其信号途径.方法取健康产妇分娩后12h内的脐带分离人脐静脉内皮细胞(HUVEC),进行细胞培养并分为5组:正常葡萄糖组(5 mmol/LD-葡萄糖,NG组);高糖组(30 mmol/LD-葡萄糖,HG组);NG+TAU组(5mmol/LD-葡萄糖+5 mmol/L牛磺酸);HG+TAU组(30 mmol/L D-葡萄糖+5 mmol/L牛磺酸);HG+TAU+Wo组(30 mmol/LD-葡萄糖+5 mmol/L牛磺酸+50 nmol/L wortmannin).干预48 h后应用流式细胞术检测细胞凋亡情况,Western印迹检测磷酸化蛋白激酶B(p-Akt)蛋白的表达水平,2,7-二氯二氢荧光素二乙酯(H2DCF-DA)探针技术及荧光倒置显微镜检测活性氧簇的相对含量.结果与NG组相比,HG组细胞的凋亡率显著增加(P＜0.05).与HG组相比,HG+TAU组的凋亡率下降(P＜0.05).与HG+TAU组相比,HG+TAU+Wo组的凋亡率较高(P＜0.05).与NG组相比,HG组磷酸化-Akt蛋白表达下降(P＜0.05).与HG组比较,HG+TAU组磷酸化-Akt蛋白表达增加(P＜0.05).与HG+TAU组相比,HG+TAU+Wo组磷酸化-Akt蛋白表达下降(P＜0.05).牛磺酸可降低高糖诱导的活性氧簇生成(P＜0.05),这一作用亦可被wortmannine所阻断(P＜0.05).结论牛磺酸通过PI3K/Akt信号途径调节细胞内活性氧簇生成,进而在高糖环境下发挥其抗凋亡作用.
목적 탐토우광산대고당유도적내피세포조망적영향급기신호도경.방법 취건강산부분면후12h내적제대분리인제정맥내피세포(HUVEC),진행세포배양병분위5조:정상포도당조(5 mmol/LD-포도당,NG조);고당조(30 mmol/LD-포도당,HG조);NG+TAU조(5mmol/LD-포도당+5 mmol/L우광산);HG+TAU조(30 mmol/L D-포도당+5 mmol/L우광산);HG+TAU+Wo조(30 mmol/LD-포도당+5 mmol/L우광산+50 nmol/L wortmannin).간예48 h후응용류식세포술검측세포조망정황,Western인적검측린산화단백격매B(p-Akt)단백적표체수평,2,7-이록이경형광소이을지(H2DCF-DA)탐침기술급형광도치현미경검측활성양족적상대함량.결과 여NG조상비,HG조세포적조망솔현저증가(P＜0.05).여HG조상비,HG+TAU조적조망솔하강(P＜0.05).여HG+TAU조상비,HG+TAU+Wo조적조망솔교고(P＜0.05).여NG조상비,HG조린산화-Akt단백표체하강(P＜0.05).여HG조비교,HG+TAU조린산화-Akt단백표체증가(P＜0.05).여HG+TAU조상비,HG+TAU+Wo조린산화-Akt단백표체하강(P＜0.05).우광산가강저고당유도적활성양족생성(P＜0.05),저일작용역가피wortmannine소조단(P＜0.05).결론 우광산통과PI3K/Akt신호도경조절세포내활성양족생성,진이재고당배경하발휘기항조망작용.
Objective To observe the effects and signalling pathway of taurine on high glucose induced endothelial apoptosis.Methods The umbilical cord from the healthy puerperal woman delivered in 12 hours was taken and the human umbilical vein endothelial cells (HUVECs)were isolated.HUVECs were cultured and divided into 5 groups:normal glucose group(5 mmol/L D-glucose,NG group),high glucose group (30 mmol/L D-glucose,HG group),NG+TAU group (5 mmol/L D-glucose+5 mmol/L taurine),HG+TAU group (30 mmol/L D-glucose+5 mmol/L taurine),HG+TAU+Wo group (30 mmol/L D-glucose+5 mmol/L taurine+ 50 nmol/L wortmannin).After 48 hours,the apoptosis was tested by Flow Cytometry and expression ofphosphorylated-protein kinase B(p-Akt)was examined by Western blot.At the same time,the relative content of reactive oxygen species (ROS) was tested by 2'-7'-Dichlorofluorescein diacetate (H2DCF-DA) probe technique and fluorescent inverted microscope.Results Compared with NG group,apoptosis rates in HG group were markedly enhanced (P＜0.05).Compared with HG group,the apoptosis rates in HG+TAU group were decreased(P＜0.05).Compared with HG+TAU group,apoptosis rates in HG+TAU+Wo group were increased(P＜0.05).Compared with HG group,expression of p-Akt protein in NG+TAU group was increased (P＜0.05).Compared with HG+TAU group,expression of p-Akt protein was reduced in HG+TAU+Wo group (P＜ 0.05).Taurine could decrease the production of ROS induced by hyperglycemia (P＜ 0.05),and the effects were blocked by wortmannin (P＜0.05).Conclusions Taurine can regulate the generation of intracellular ROS by PI3K/Akt pathway,then prevent apoptosis of endothelial cell induced by high glucose.