中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2012年
4期
327-331
,共5页
动脉粥样硬化%一氧化氮合酶%植物雌激素类
動脈粥樣硬化%一氧化氮閤酶%植物雌激素類
동맥죽양경화%일양화담합매%식물자격소류
Atherosclerosis%Nitric oxide synthase%Phytoestrogens
目的 探讨磷酸肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路在植物雌激素染料木素(genistein)促进内皮型一氧化氮合酶(eNOS)活性过程中的作用.方法 体外培养人脐静脉内皮细胞(HUVEC),在氧化型低密度脂蛋白(ox-LDL,100 mg/L)以及ox-LDL( 100 mg/L)加genistein( 100nmoL/L)分别作用5、10、15、30和60 min的基础上,Greiss反应测定细胞培养上清中一氧化氮(N0)的含量,RT-PCR检测HUVEC eNOS mRNA的表达,Western blot检测HUVEC eNOS蛋白和磷酸化eNOS (Ser1179)的表达水平.Western blot检测PI3K/AKT抑制剂LY294002和NSC154020干预后,genistein 对HUVEC磷酸化eNOS( Ser1179)表达的影响.结果 HUVEC经100 nmol/L genistein(5、10、15、30和60 min)处理后,培养液NO浓度和磷酸化eNOS(Ser1179)表达水平均明显高于ox-LDL组(P均＜0.05),其中genistein 15 min处理组最高,然而eNOS mRNA和非磷酸化eNOS蛋白表达水平与ox-LDL 组比较差异无统计学意义(P＞0.05).HUVEC经PI3K/AKT信号通路抑制剂LY294002和NSC154020干预后磷酸化eNOS( Ser1179)表达明显低于genistein 15 min处理组(P＜0.05).结论 genistein对HUVEC NO的诱导作用与其促进eNOS(Ser1175)磷酸化进而提高eNOS活性密切相关,而genistein对磷酸化eNOS( Ser1179)的诱导作用被PI3K/AKT抑制剂LY294002和NSC154020阻断,可见genistein在促进eNOS活性过程中通过PI3K/AKT信号通路发挥重要作用.
目的 探討燐痠肌醇3激酶/蛋白激酶B(PI3K/AKT)信號通路在植物雌激素染料木素(genistein)促進內皮型一氧化氮閤酶(eNOS)活性過程中的作用.方法 體外培養人臍靜脈內皮細胞(HUVEC),在氧化型低密度脂蛋白(ox-LDL,100 mg/L)以及ox-LDL( 100 mg/L)加genistein( 100nmoL/L)分彆作用5、10、15、30和60 min的基礎上,Greiss反應測定細胞培養上清中一氧化氮(N0)的含量,RT-PCR檢測HUVEC eNOS mRNA的錶達,Western blot檢測HUVEC eNOS蛋白和燐痠化eNOS (Ser1179)的錶達水平.Western blot檢測PI3K/AKT抑製劑LY294002和NSC154020榦預後,genistein 對HUVEC燐痠化eNOS( Ser1179)錶達的影響.結果 HUVEC經100 nmol/L genistein(5、10、15、30和60 min)處理後,培養液NO濃度和燐痠化eNOS(Ser1179)錶達水平均明顯高于ox-LDL組(P均＜0.05),其中genistein 15 min處理組最高,然而eNOS mRNA和非燐痠化eNOS蛋白錶達水平與ox-LDL 組比較差異無統計學意義(P＞0.05).HUVEC經PI3K/AKT信號通路抑製劑LY294002和NSC154020榦預後燐痠化eNOS( Ser1179)錶達明顯低于genistein 15 min處理組(P＜0.05).結論 genistein對HUVEC NO的誘導作用與其促進eNOS(Ser1175)燐痠化進而提高eNOS活性密切相關,而genistein對燐痠化eNOS( Ser1179)的誘導作用被PI3K/AKT抑製劑LY294002和NSC154020阻斷,可見genistein在促進eNOS活性過程中通過PI3K/AKT信號通路髮揮重要作用.
목적 탐토린산기순3격매/단백격매B(PI3K/AKT)신호통로재식물자격소염료목소(genistein)촉진내피형일양화담합매(eNOS)활성과정중적작용.방법 체외배양인제정맥내피세포(HUVEC),재양화형저밀도지단백(ox-LDL,100 mg/L)이급ox-LDL( 100 mg/L)가genistein( 100nmoL/L)분별작용5、10、15、30화60 min적기출상,Greiss반응측정세포배양상청중일양화담(N0)적함량,RT-PCR검측HUVEC eNOS mRNA적표체,Western blot검측HUVEC eNOS단백화린산화eNOS (Ser1179)적표체수평.Western blot검측PI3K/AKT억제제LY294002화NSC154020간예후,genistein 대HUVEC린산화eNOS( Ser1179)표체적영향.결과 HUVEC경100 nmol/L genistein(5、10、15、30화60 min)처리후,배양액NO농도화린산화eNOS(Ser1179)표체수평균명현고우ox-LDL조(P균＜0.05),기중genistein 15 min처리조최고,연이eNOS mRNA화비린산화eNOS단백표체수평여ox-LDL 조비교차이무통계학의의(P＞0.05).HUVEC경PI3K/AKT신호통로억제제LY294002화NSC154020간예후린산화eNOS( Ser1179)표체명현저우genistein 15 min처리조(P＜0.05).결론 genistein대HUVEC NO적유도작용여기촉진eNOS(Ser1175)린산화진이제고eNOS활성밀절상관,이genistein대린산화eNOS( Ser1179)적유도작용피PI3K/AKT억제제LY294002화NSC154020조단,가견genistein재촉진eNOS활성과정중통과PI3K/AKT신호통로발휘중요작용.
Objective Genistein could inhibit the development of atherosclerosis. This study explored the role of PI3K/AKT signaling during genistein promoted eNOS activation.Methods Human umbilical vein endothelial cells (HUVECs) were incubated with ox-LDL( 100 mg/L),then treated with genistein (100 nmol/L) for 5,10,15,30 and 60 min.The production of NO was assessed by Griess reaction in cell culture supernatant.The mRNA expression of endothelial nitric oxide synthase (eNOS) was detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein expression of eNOS and phosphorylation eNOS(Ser1179)were determined by Western blot.The effect of genistein on phosphorylation eNOS( Ser1179 ) level was also observed in the presence of LY294002 or NSC154020 ( PI3K and AKT inhibitors).Results The concentration of NO and the expression level of phosphorylation eNOS ( Ser1179 )were significantly increased in ox-LDL + genistein treated cells than ox-LDL treated cells ( all P＜0.05 ),and the peak effects were observed at 15 min,however,eNOS mRNA and non-phosphorylated eNOS protein expression were similar between the two groups ( P＞0.05 ). Furthermore,the expression level of phosphorylation eNOS(Ser1179 ) was significantly lower in PIK3/AKT inhibitors LY294002 and NSC154020 treated cells compared with ox-LDL + genistein treated cells (all P＜0.05 ).Conclusion Genistein could promote the activity of eNOS through increasing phosphorylation eNOS(Ser1179) level through PI3K/AKT pathway.
