卡托普利晚期预处理对人内皮细胞缺氧复氧损伤时内皮素1和一氧化氮合酶表达的影响

Effects of Catopril on Expressions of Edothelin-1 and Nitric Oxide Synthase against the Anoxia-Reoxygenation Injury to Endothelial Cells

观察缺氧复氧损伤时内皮细胞表达内皮素、一氧化氮合酶和一氧化氮的变化 ,并探讨卡托普利晚期预处理对内皮细胞在缺氧复氧损伤时三者的影响及机制。选用体外培养的第 3代人脐静脉内皮细胞 ,设正常对照组、单纯缺氧组、缺氧复氧组、卡托普利预处理组、卡托普利 +缓激肽B2 受体阻断剂组、卡托普利 +蛋白激酶C阻断剂组和卡托普利 +核因子κB阻断剂组 ,然后提取总RNA ,运用半定量逆转录—聚合酶链反应检测内皮素、一氧化氮合酶的mRNA表达。并利用分光光度计检测总一氧化氮合酶和一氧化氮蛋白的表达。结果发现 ,内皮素的灰度值在单纯缺氧组、缺氧复氧组均升高 ,在卡托普利组降低 ,而在三种阻断剂组均升高 ;诱导型一氧化氮合酶和内皮型一氧化氮合酶灰度值的变化规律与内皮素相反 ,其中诱导型一氧化氮合酶的变化较为轻微。与对照组相比 ,单纯缺氧组和缺氧复氧组一氧化氮合酶和一氧化氮有不同程度的降低 (P <0 .0 1) ;与单纯缺氧组和缺氧复氧组相比 ,卡托普利组一氧化氮合酶和一氧化氮均明显升高 (P <0 .0 1) ;与卡托普利组相比 ,3种阻断剂均可使一氧化氮合酶和一氧化氮的表达下降 (P <0 .0 1) ,而与缺氧复氧组相比表达升高 (P <0 .0 1)。结果提示 ,缺氧复氧可以使内皮细胞表达的内皮素增加 ,一氧化氮合酶降低 ,

Aim To observe the expressions of endothelin-1 (ET-1), nitric oxide (NO) and nitric oxide synthase (NOS) in the anoxia-reoxygenation (H/R) injury on human umbilical vein endothelial cells (EC), and further research the molecular mechanism of catopril late effect on production of ET-1 and NO. Methods The third passage of cultured EC was randomly divided into 7 groups: control, anoxia, anoxia-reoxygenation, catopril, catopril+bradykinin Β 2 receptor inhibitor, catopril+PKC inhibitor, catopril+NF-κB inhibitor. Total RNA was extracted. The expression of ET-1, endthelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) mRNA were analyzed by semi-quantiative RT-PCR method. The protein level of total NOS and NO were analyzed too. Results ET-1 mRNA expression increased significantly in anoxia group and anoxia-reoxygenation group, decreased in catopril group, and increased in three inhibitor groups. While the mRNA expressions of iNOS and eNOS were contrary to that of ET-1, and the change of iNOS was slightly. The protein levels of total NOS and NO are lower in anoxia group and anoxia-reoxygenation group than in control group, higher in catopril group than in anoxia group or anoxia-reoxygenation group, and lower in three inhibitor groups than in catopril group, but higher than in anoxia-reoxygenation group. Conclusion Anoxia-reoxygenation injury can induce ET-1 expression increasing and NO expression decreasing. Catopril can protect the imbalance of ET-1 and NO. Bradykinin Β 2 receptor, PKC activating and nucleus factor are partly involved in the protective effect.