中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2009年
5期
436-440
,共5页
文渊%马业新%张新金%洪李锋%冯达应%卢振华
文淵%馬業新%張新金%洪李鋒%馮達應%盧振華
문연%마업신%장신금%홍리봉%풍체응%로진화
红细胞生成素%信号传导%血管紧张素Ⅱ
紅細胞生成素%信號傳導%血管緊張素Ⅱ
홍세포생성소%신호전도%혈관긴장소Ⅱ
Erythropoietin%Signal transduction%Angiotensin Ⅱ
目的 探讨促红细胞生成素(EPO)对血管紧张素Ⅱ(AngⅡ)诱导的肥大心肌细胞的影响,以及磷脂酰肌醇3激酶(PI3K)/丝氨酸苏氨酸激酶(Akt)-内皮型一氧化氮合酶(eNOS)信号转导通路在其中的作用.方法 分离乳鼠心肌细胞,利用AngⅡ诱导建立心肌细胞肥大模型,以心肌细胞表面积和心钠素(ANF)mRNA表达作为心肌细胞肥大观察指标.观察不同浓度EPO对肥大心肌细胞的影响,并利用PI3K抑制剂LY294002和一氧化氮合酶抑制剂L-NAME对其相关机制进行探讨,I司时对细胞培养液中一氧化氮(NO)浓度进行检测,蛋白免疫印迹法检测磷酸化Akt(p-Akt)、Akt、磷酸化eNOS(p-eNOS)和eNOS蛋白表达情况.结果 20 U/ml EPO能抑制由AngⅡ诱导的心肌细胞肥大,表现为心肌细胞表面积和ANF mRNA表达均减少(P<0.05).EPO能激活Akt,促进eNOS及p-eNOS表达增加(均P<0.05),并使NO合成增加(P<0.01).LY294002和L-NAME能逆转EPO的抗心肌细胞肥大作用,减少NO产最(P<0.05).蛋白免疫印迹法榆测显示,LY294002能够抑制EPO对p-Akt、p-eNOS和eNOS蛋白表达的促进作用,而L-NAME能抑制eNOS的磷酸化(均P<0.05).结论 EPO能够抑制AngⅡ诱导的心肌细胞肥大,该作用可能是通过激活P13K/Akt信号转导通路,促进eNOS表达与活化,从而促进NO的合成来实现的.
目的 探討促紅細胞生成素(EPO)對血管緊張素Ⅱ(AngⅡ)誘導的肥大心肌細胞的影響,以及燐脂酰肌醇3激酶(PI3K)/絲氨痠囌氨痠激酶(Akt)-內皮型一氧化氮閤酶(eNOS)信號轉導通路在其中的作用.方法 分離乳鼠心肌細胞,利用AngⅡ誘導建立心肌細胞肥大模型,以心肌細胞錶麵積和心鈉素(ANF)mRNA錶達作為心肌細胞肥大觀察指標.觀察不同濃度EPO對肥大心肌細胞的影響,併利用PI3K抑製劑LY294002和一氧化氮閤酶抑製劑L-NAME對其相關機製進行探討,I司時對細胞培養液中一氧化氮(NO)濃度進行檢測,蛋白免疫印跡法檢測燐痠化Akt(p-Akt)、Akt、燐痠化eNOS(p-eNOS)和eNOS蛋白錶達情況.結果 20 U/ml EPO能抑製由AngⅡ誘導的心肌細胞肥大,錶現為心肌細胞錶麵積和ANF mRNA錶達均減少(P<0.05).EPO能激活Akt,促進eNOS及p-eNOS錶達增加(均P<0.05),併使NO閤成增加(P<0.01).LY294002和L-NAME能逆轉EPO的抗心肌細胞肥大作用,減少NO產最(P<0.05).蛋白免疫印跡法榆測顯示,LY294002能夠抑製EPO對p-Akt、p-eNOS和eNOS蛋白錶達的促進作用,而L-NAME能抑製eNOS的燐痠化(均P<0.05).結論 EPO能夠抑製AngⅡ誘導的心肌細胞肥大,該作用可能是通過激活P13K/Akt信號轉導通路,促進eNOS錶達與活化,從而促進NO的閤成來實現的.
목적 탐토촉홍세포생성소(EPO)대혈관긴장소Ⅱ(AngⅡ)유도적비대심기세포적영향,이급린지선기순3격매(PI3K)/사안산소안산격매(Akt)-내피형일양화담합매(eNOS)신호전도통로재기중적작용.방법 분리유서심기세포,이용AngⅡ유도건립심기세포비대모형,이심기세포표면적화심납소(ANF)mRNA표체작위심기세포비대관찰지표.관찰불동농도EPO대비대심기세포적영향,병이용PI3K억제제LY294002화일양화담합매억제제L-NAME대기상관궤제진행탐토,I사시대세포배양액중일양화담(NO)농도진행검측,단백면역인적법검측린산화Akt(p-Akt)、Akt、린산화eNOS(p-eNOS)화eNOS단백표체정황.결과 20 U/ml EPO능억제유AngⅡ유도적심기세포비대,표현위심기세포표면적화ANF mRNA표체균감소(P<0.05).EPO능격활Akt,촉진eNOS급p-eNOS표체증가(균P<0.05),병사NO합성증가(P<0.01).LY294002화L-NAME능역전EPO적항심기세포비대작용,감소NO산최(P<0.05).단백면역인적법유측현시,LY294002능구억제EPO대p-Akt、p-eNOS화eNOS단백표체적촉진작용,이L-NAME능억제eNOS적린산화(균P<0.05).결론 EPO능구억제AngⅡ유도적심기세포비대,해작용가능시통과격활P13K/Akt신호전도통로,촉진eNOS표체여활화,종이촉진NO적합성래실현적.
Objective To explore the effect of erythropoietin (EPO) on angiotensin Ⅱ (Ang Ⅱ) induced neonatal rat cardiomyocyte hypertrophy and the association with PI3K/Akt-eNOS signaling pathway. Methods Cardiomyocytes were isolated from new-born Sprague-Dawley rats and stimulated by Ang Ⅱ in vitro. The cell surface area and mRNA expression of atrial natriuretic factor (ANF) of cardiomyocytes were determined in the presence and absence of various concentrations of EPO, phosphatidylinositol 3'-kinase (PI3K) inhibitor LY294002 and nitric oxide synthase (NOS) inhibitor L-NAME. Intracellular signal molecules, such as Akt, phosphorylated Akt, eNOS and phosphorylated eNOS protein expressions were detected by western blot. Nitric oxide (NO) level in the supernatant of cultured cardiomyocytes was assayed by NO assay kit. Results EPO (20 U/ml) significantly inhibited Ang Ⅱ induced cardiomyocyte hypertrophy as shown by decreased cell surface area and ANF mRNA expression (all P <0.05). EPO also activated Akt and enhanced the expression of eNOS and its phosphorylation (all P < 0.05), increased the NO production (P <0.01). These effects could be partially abolished by cotreatment with LY294002 or L-NAME (all P < 0.05). Conclusion EPO attenuates Ang Ⅱ induced cardiomyocytes hypertrophy via activating PI3K-Akt-eNOS pathway and promoting NO production.